234 results on '"Wool IG"'
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2. Accumulation of amino acids in muscle of perfused rat heart. Effect of insulin
- Author
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Scharff, R and Wool, IG
- Abstract
1. Rat heart perfused with Krebs-Henseleit bicarbonate buffer released material containing ninhydrin-positive nitrogen, but the amount was less than that reported to be released by diaphragm; glucose, but not insulin, decreased the release of ninhydrin-positive nitrogen and increased the concentration of the same material in the intracellular water of heart. 2. When heart was perfused with a mixture of amino acids and glucose, there was actually a net uptake, and an increase in intracellular concentration, of ninhydrin-positive nitrogen. Changes in the concentration of ninhydrin-positive nitrogen did not accurately reflect changes in concentration of amino acids. 3. The effect of insulin on the actual concentration of individual amino acids in heart muscle was examined by perfusing the heart with a mixture of amino acids and other ninhydrin-positive substances in the same concentration as they are found in plasma. 4. The effect of insulin on the concentrations of amino acids in the medium and in the intracellular water of the heart was determined after perfusion for different periods of time. No clear or meaningful effect of insulin was observed, despite the fact that insulin significantly increased the accumulation, in each of the same hearts, of radioactivity from amino[(14)C]isobutyric acid.
- Published
- 1965
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3. INSULIN AND INCORPORATION OF AMINO ACIDS INTO PROTEIN OF MUSCLE. 2. ACCUMULATION AND INCORPORATION STUDIES WITH THE PERFUSED RAT HEART
- Author
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MANCHESTER, KL and WOOL, IG
- Published
- 1963
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4. The identification of the determinants of the cyclic, sequential binding of elongation factors tu and g to the ribosome.
- Author
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Yu H, Chan YL, and Wool IG
- Subjects
- Base Sequence, Binding Sites, Escherichia coli chemistry, Models, Molecular, Molecular Sequence Data, Nucleic Acid Conformation, Protein Binding, RNA, Bacterial chemistry, RNA, Bacterial metabolism, RNA, Ribosomal, 23S chemistry, RNA, Ribosomal, 23S metabolism, Ribosome Subunits, Large, Bacterial chemistry, Escherichia coli metabolism, Peptide Elongation Factor G metabolism, Peptide Elongation Factor Tu metabolism, Ribosomes chemistry, Ribosomes metabolism
- Abstract
Experiments dedicated to gaining an understanding of the mechanism underlying the orderly, sequential association of elongation factor Tu (EF-Tu) and elongation factor G (EF-G) with the ribosome during protein synthesis were undertaken. The binding of one EF is always followed by the binding of the other, despite the two sharing the same-or a largely overlapping-site and despite the two having isosteric structures. Aminoacyl-tRNA, peptidyl-tRNA, and deacylated-tRNA were bound in various combinations to the A-site, P-site, or E-site of ribosomes, and their effect on conformation in the peptidyl transferase center, the GTPase-associated center, and the sarcin/ricin domain (SRD) was determined. In addition, the effect of the ribosome complexes on sensitivity to the ribotoxins sarcin and pokeweed antiviral protein and on the binding of EF-G*GTP were assessed. The results support the following conclusions: the EF-Tu ternary complex binds to the A-site whenever it is vacant and the P-site has peptidyl-tRNA; and association of the EF-Tu ternary complex is prevented, simply by steric hindrance, when the A-site is occupied by peptidyl-tRNA. On the other hand, the affinity of the ribosome for EF-G*GTP is increased when peptidyl-tRNA is in the A-site, and the increase is the result of a conformational change in the SRD. We propose that peptidyl-tRNA in the A-site is an effector that initiates a series of changes in tertiary interactions between nucleotides in the peptidyl transferase center, the SRD, and the GTPase-associated center of 23S rRNA; and that the signal, transmitted through a transduction pathway, informs the ribosome of the position of peptidyl-tRNA and leads to a conformational change in the SRD that favors binding of EF-G.
- Published
- 2009
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5. The integrity of the sarcin/ricin domain of 23 S ribosomal RNA is not required for elongation factor-independent peptide synthesis.
- Author
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Chan YL and Wool IG
- Subjects
- Base Sequence, Molecular Sequence Data, Peptide Elongation Factors metabolism, Protein Synthesis Inhibitors pharmacology, RNA, Ribosomal, 23S drug effects, Ribosomes drug effects, Ricin pharmacology, Toxins, Biological pharmacology, Nucleic Acid Conformation drug effects, Protein Biosynthesis drug effects, RNA, Ribosomal, 23S chemistry, RNA, Ribosomal, 23S metabolism, Ribosomes metabolism
- Abstract
The elongation stage of protein synthesis consists of repeated cycles of the binding of aminoacyl-tRNA, peptide bond formation, and translocation. The process is normally catalyzed by the elongation factors Tu and G; however, the reactions can proceed, at least in prescribed and limited circumstance, in the absence of the elongation factors, a finding that strongly implies that the chemistry of protein synthesis is inherent in the ribosome. The sarcin/ricin domain in 23 S rRNA, the site of inactivation of ribosomes by ribotoxins, is where the elongation factors bind. The question that arises is whether the sarcin/ricin domain is necessary for factor-independent peptide synthesis. The answer is that it is not. The disruption of the sarcin/ricin domain by covalent modification with either sarcin or pokeweed antiviral protein did not affect factor-independent peptide synthesis; nor did lethal mutations of nucleotides that abolish the binding of elongation factors. The results imply that the sole function of the sarcin/ricin domain is to provide a binding site for the elongation factors and, hence, to facilitate the elongation reactions. The results also raise the possibility of the co-evolution of the sarcin/ricin domain and the elongation factors.
- Published
- 2008
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6. Determination of the amino acids in yeast ribosomal protein YS11 essential for the recognition of nucleotides in 18 S ribosomal RNA.
- Author
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Dresios J, Chan YL, and Wool IG
- Subjects
- Amino Acid Sequence, Amino Acid Substitution, Amino Acids, Base Sequence, Binding Sites, Gene Expression Regulation, Fungal, Molecular Biology, Mutation, Protein Binding, Protein Conformation, RNA, Ribosomal, 18S chemistry, Ribosomal Proteins chemistry, Ribosomal Proteins genetics, Saccharomyces cerevisiae Proteins chemistry, Saccharomyces cerevisiae Proteins genetics, RNA, Ribosomal, 18S metabolism, Ribosomal Proteins metabolism, Saccharomyces cerevisiae metabolism, Saccharomyces cerevisiae Proteins metabolism
- Abstract
The nucleotides in domain I of 18 S rRNA that are important for the binding of the essential yeast ribosomal protein YS11 are mainly in a kink-turn motif and the terminal loop of helix 11 (H11). In the atomic structure of the Thermus thermophilus 30 S subunit, 16 amino acids in S17, the homolog of YS11, are within hydrogen bonding distance of nucleotides in 16 S rRNA. The homologous or analogous 16 amino acids in YS11 were replaced with alanine; nine of the substitutions slowed the growth of yeast cells. The most severe effects were caused by mutations R103A, N106A, K133A, T134A, and K151A. The T. thermophilus analogs of Arg103, Asn106, Thr134, and Lys151 contact nucleotides in the kink-turn motif of 16 S rRNA, whereas Lys133 contacts nucleotides in the terminal loop of H11. These contacts are predominantly with backbone phosphate and sugar oxygens in regions that deviate from A-form geometry, suggesting that YS11 recognizes the shape of its rRNA-binding site rather than reading the sequence of nucleotides. The effect of the mutations on the binding of YS11 to a domain I fragment of 18 S rRNA accorded, in general, with their effect on growth. Mutations of seven YS11 amino acids (Ser77, Met80, Arg88, Tyr97, Pro130, Ser132, and Arg136) whose homologs or analogs in S17 are within hydrogen bonding distance of nucleotides in 16 S rRNA did not affect binding. Apparently, proximities alone do not define either the amino acids or the nucleotides that are important for recognition.
- Published
- 2006
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7. A pathway for the transmission of allosteric signals in the ribosome through a network of RNA tertiary interactions.
- Author
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Chan YL, Dresios J, and Wool IG
- Subjects
- Allosteric Regulation, Models, Molecular, Mutation, Protein Subunits genetics, Protein Subunits metabolism, Ribosomes genetics, Nucleic Acid Conformation, RNA, Ribosomal chemistry, RNA, Ribosomal metabolism, Ribosomes metabolism, Signal Transduction physiology
- Abstract
There are a large number of tertiary contacts between nucleotides in 23S rRNA, but which are of functional importance is not known. Disruption of one between A2662 in the sarcin/ricin loop (SRL) and A2531 in the peptidyl-transferase center (PTC) has adverse effects on cell growth and on the ability of ribosomes to catalyze some but not other partial reactions of elongation. A lethal A2662C mutation is suppressed by a concomitant lethal A2531 mutation. Ribosomes with non-lethal A2531 mutations, treated with base-specific reagents, have alterations of nucleotides in the PTC (home of A2531) and, more significantly, in nucleotides in the SRL and in the GTPase center. The results suggest that the function of ribosomal centers is coordinated by a set of sequential conformational changes in rRNA that are a response to signals transmitted through a network of tertiary interactions.
- Published
- 2006
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8. A determination of the identity elements in yeast 18 S ribosomal RNA for the recognition of ribosomal protein YS11: the role of the kink-turn motif in helix 11.
- Author
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Dresios J, Chan YL, and Wool IG
- Subjects
- Amino Acid Sequence, Base Pairing, Base Sequence, Binding Sites, Models, Molecular, Molecular Sequence Data, Mutation genetics, Protein Conformation, RNA, Ribosomal, 18S genetics, Ribosomal Proteins chemistry, Saccharomyces cerevisiae chemistry, Saccharomyces cerevisiae genetics, Sequence Alignment, Nucleic Acid Conformation, RNA, Ribosomal, 18S chemistry, RNA, Ribosomal, 18S metabolism, Ribosomal Proteins metabolism, Saccharomyces cerevisiae metabolism
- Abstract
A description of the site of interaction of YS11, the yeast homolog of eubacterial S17, with 18 S rRNA was obtained by assessing the binding of the ribosomal protein, in a filter retention assay, to oligoribonucleotides that reproduce regions of 18 S rRNA. YS11 binds predominantly to domain I; the Kd value is 113 nM. The dimensions of the YS11 binding site were refined, guided by chemical protection data and by the atomic structure of the Thermus thermophilus 30 S subunit, which has the S17 recognition site in 16 S rRNA. An oligoribonucleotide that mimics helix 11, a phylogenetically conserved region in domain I, binds YS11 with a Kd value of 230 nM; a second oligoribonucleotide that contains only the kink-turn motif in helix 11 binds YS11 with a Kd value of 528 nM. Thus, helix 11 has most of the nucleotides required for the recognition of YS11. To identify those nucleotides a set of 27 transversion mutations in H11 was constructed and their contribution to the binding of YS11 determined. Mutations of nine nucleotides (U313, C314, A316, G337, C338, G347, U348, U350, and C351) increased the Kd value for YS11 binding by at least eightfold; G325U and U349A mutations increased the Kd value fivefold. Eight of the 11 mutations are in the kink-turn in H11, confirming the critical importance of the motif for YS11 recognition. The other three nucleotides are in the lower stem and the terminal loop of H11, which makes a lesser, but still important, contribution to YS11 binding. The identity elements for YS11 recognition are: A316, G325, G337, G347, U348, U349, U350, and C351. The effect of the other nucleotides that decrease binding is probably indirect, presumably they affect the conformation of the binding site but do not have contacts to YS11 amino acid residues. The eight identity element nucleotides are in regions of H11 that deviate from A-form geometry and the contacts are predominantly, if not exclusively, to backbone phosphate and sugar oxygen atoms, indicating that YS11 recognizes the shape of the rRNA binding site rather than reading the sequence of nucleotides.
- Published
- 2005
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9. The location and the significance of a cross-link between the sarcin/ricin domain of ribosomal RNA and the elongation factor-G.
- Author
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Chan YL, Correll CC, and Wool IG
- Subjects
- Amino Acid Sequence, Bacterial Proteins chemistry, Bacterial Proteins genetics, Bacterial Proteins metabolism, Base Sequence, Binding Sites genetics, Cross-Linking Reagents, Escherichia coli genetics, Escherichia coli metabolism, Escherichia coli Proteins chemistry, Escherichia coli Proteins genetics, Escherichia coli Proteins metabolism, Macromolecular Substances, Models, Molecular, Molecular Sequence Data, Nucleic Acid Conformation, RNA, Bacterial chemistry, RNA, Bacterial genetics, RNA, Bacterial metabolism, RNA, Ribosomal genetics, Sequence Homology, Amino Acid, Thermus thermophilus genetics, Thermus thermophilus metabolism, Ultraviolet Rays, Peptide Elongation Factor G chemistry, Peptide Elongation Factor G metabolism, RNA, Ribosomal chemistry, RNA, Ribosomal metabolism, Ricin chemistry, Ricin metabolism
- Abstract
During translocation peptidyl-tRNA moves from the A-site to the P-site and mRNA is displaced by three nucleotides in the 3' direction. This reaction is catalyzed by elongation factor-G (EF-G) and is associated with ribosome-dependent hydrolysis of GTP. The molecular basis of translocation is the most important unsolved problem with respect to ribosome function. A critical question, one that might provide a clue to the mechanism of translocation, is the precise identity of the contacts between EF-G and ribosome components. To make the identification, a covalent bond was formed, by ultraviolet irradiation, between EF-G and a sarcin/ricin domain (SRD) oligoribonucleotide containing 5-iodouridine. The cross-link was established, by mass spectroscopy and by Edman degradation, to be between a tryptophan at position 127 in the G domain in EF-G and either one of two 5-iodouridine nucleotides in the sequence UAG2655U in the SRD. G2655 is a critical identity element for the recognition of the factor's ribosomal binding site. The site of the cross-link provides the first direct evidence that the SRD is in close proximity to the EF-G catalytic center. The proximity suggests that the SRD RNA has a role in the activation of GTP hydrolysis that leads to a transition in the conformation of the factor and to its release from the ribosome.
- Published
- 2004
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10. The role of the zinc finger motif and of the residues at the amino terminus in the function of yeast ribosomal protein YL37a.
- Author
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Dresios J, Chan YL, and Wool IG
- Subjects
- Amino Acid Motifs, Amino Acid Sequence, Amino Acids, Aromatic genetics, Amino Acids, Aromatic metabolism, Amino Acids, Basic genetics, Amino Acids, Basic metabolism, Archaeal Proteins chemistry, Archaeal Proteins metabolism, Binding Sites, Haloarcula marismortui genetics, Magnesium pharmacology, Models, Molecular, Molecular Sequence Data, Mutation genetics, Nucleic Acid Conformation, Phenotype, Protein Binding, RNA, Ribosomal chemistry, RNA, Ribosomal genetics, RNA, Ribosomal metabolism, RNA, Ribosomal, 23S chemistry, RNA, Ribosomal, 23S genetics, RNA, Ribosomal, 23S metabolism, RNA-Binding Proteins chemistry, RNA-Binding Proteins genetics, RNA-Binding Proteins metabolism, Recombinant Fusion Proteins chemistry, Recombinant Fusion Proteins metabolism, Ribosomal Proteins genetics, Saccharomyces cerevisiae Proteins genetics, Structure-Activity Relationship, Ribosomal Proteins chemistry, Ribosomal Proteins metabolism, Saccharomyces cerevisiae genetics, Saccharomyces cerevisiae growth & development, Saccharomyces cerevisiae Proteins chemistry, Saccharomyces cerevisiae Proteins metabolism, Zinc Fingers
- Abstract
YL37a is an essential yeast ribosomal protein that has a C(2)-C(2) zinc finger motif. Replacement of the cysteine residues had yielded variants that lacked the capacity to bind zinc but still supported cell growth. In a continuation of an examination of the relation of the structure of YL37a to its function, the contribution of amino acid residues in the intervening sequence between the internal cysteine residues of the motif was evaluated. Substitutions of alanine for the lysine residues at positions 44, 45, or 48, or for arginine 49 slowed cell growth. The most severe effect was caused by a double-mutation, K48A-R49A. A mutation of tryptophan 55 to alanine was lethal. Mutations to alanine of six conserved residues (K6, K7, K13, Y14, R17, and Y18) in the amino-terminal region decreased cell growth; the Y14 mutation was lethal. An in vitro assay for binding of YL37a to individual 26 S rRNA domains was developed. Binding of the recombinant fusion protein MBP-YL37a was to domains II and III; the K(d) for binding to domain II was 79 nM; for domain III it was 198 nM. There was a close correspondence between the effect of mutations in YL37a on cell growth and on binding to 26 S rRNA. In the atomic structure of the 50 S subunit of Haloarcula marismortui, the archaebacteria homolog of yeast YL37a, L37ae, coordinates a zinc atom and the finger motif is folded and interacts mainly with domain III of 23 S rRNA; whereas the amino-terminal region of L37ae interacts primarily with domain II. The biochemical and genetic experiments complement the three-dimensional structure and define for the first time the functional importance of a subset of the residues in close proximity to nucleotides., (Copyright 2002 Elsevier Science Ltd.)
- Published
- 2002
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11. Analysis by systematic deletion of amino acids of the action of the ribotoxin restrictocin.
- Author
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Glück A and Wool IG
- Subjects
- Amino Acid Sequence, Amino Acids genetics, Animals, Antigens, Plant, Cytotoxins genetics, DNA, Complementary chemistry, Fungal Proteins genetics, Gene Deletion, Lysine chemistry, Molecular Sequence Data, Mutation, Plasmids, Protein Structure, Secondary, RNA, Ribosomal, 28S chemistry, Rabbits, Rats, Reticulocytes chemistry, Ribonucleases genetics, Allergens, Cytotoxins chemistry, Fungal Proteins chemistry, Ribonucleases chemistry, Ribosomes chemistry
- Abstract
A series of contiguous deletions were made in a cDNA encoding the ribonuclease restrictocin with the purpose of identifying the amino acids that are essential for the cleavage of the phosphodiester bond on the 3' side of G4325 in the alpha-sarcin/ricin domain of mammalian (rat) 28S rRNA. In all 93 of 149 amino acids, 62% of the residues in restrictocin, were not essential for the action of the toxin. Of the five residues that have been proposed to constitute the active site, three could be deleted without loss of activity if they were part of a deletion of three or five amino acids but not if they were removed singly. It is likely that the loss of these three residues is compensated for by a neighboring residue that occupies the structural space created by the larger amino acid deletions. This was demonstrated to be the case for the active site residue Glu95 which in the deletion mutant Delta91-95 is replaced by Asp90. Systematic deletion of amino acids is a rapid, cost effective method for identifying the residues in a protein likely to contribute directly to function and, hence, deserving of closer scrutiny. Moreover, a semiquantitative estimate of the contribution of the residue to function can be made. For this reason the method may be useful for functional proteomics.
- Published
- 2002
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12. The phenotype of mutations of the base-pair C2658.G2663 that closes the tetraloop in the sarcin/ricin domain of Escherichia coli 23 S ribosomal RNA.
- Author
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Chan YL, Sitikov AS, and Wool IG
- Subjects
- Bacterial Proteins biosynthesis, Bacterial Proteins metabolism, Base Sequence, Binding Sites, Escherichia coli growth & development, Genes, Lethal genetics, Guanosine Triphosphate metabolism, Oligoribonucleotides chemistry, Oligoribonucleotides genetics, Oligoribonucleotides metabolism, Peptide Elongation Factor G metabolism, Peptides metabolism, Phenotype, Plasmids genetics, Promoter Regions, Genetic genetics, Protein Binding, Protein Biosynthesis genetics, RNA, Bacterial chemistry, RNA, Bacterial genetics, RNA, Bacterial metabolism, RNA, Ribosomal, 23S genetics, RNA, Ribosomal, 23S metabolism, Ribosomes genetics, Ribosomes metabolism, Thermodynamics, rRNA Operon genetics, Base Pairing genetics, Endoribonucleases metabolism, Escherichia coli genetics, Fungal Proteins, Mutation genetics, RNA, Ribosomal, 23S chemistry, Ricin metabolism
- Abstract
The sarcin/ricin domain (SRD) in Escherichia coli 23 S rRNA is a part of the site for the association of elongation factors with ribosomes and for that reason is critical for the binding of aminoacyl-tRNA and for translocation during the reiterative elongation reactions of protein synthesis. The SRD has a GAGA tetraloop that is shut off by a Watson-Crick C2658 x G2663 pair. The contribution of this pair to the function of the ribosome has been evaluated by constructing mutations in the nucleotides and determining their phenotype. Constitutive expression of a plasmid-encoded rrnB operon with a G2663C transversion mutation that disrupts the Watson-Crick pair was lethal. Double transversion mutations, C2658G x G2663C and C2658A x G2663U, that reverse the polarity of the pyrimidine and the purine but restore the potential to form a canonical pair, were also lethal. Induction of transcription of 23 S rRNA with the same mutations, but encoded in a plasmid with a lambdaP(L) promoter and expressed at a lower level, retarded growth. The sedimentation profiles of ribosomes with transversion mutations in C2658 and/or G2663 are altered; the ratio of 50 S subunits to 30 S particles is changed and polysomes are reduced. Ribosomes with a G2663C, a C2658G x G2663C, or a C2658A x G2663U mutation in 23 S rRNA were not active in protein synthesis, indeed, they appeared to inhibit the activity of ribosomes with wild-type 23 S rRNA. Transversion mutations in the analogs of C2658 and G2663 decreased binding of EF-G to SRD oligoribonucleotides; the same mutations in 23 S rRNA decreased binding of the factor to intact ribosomes. The most severe phenotype, in growth, in protein synthesis, and in the binding of EF-G, was associated with a C2658G x G2663C mutation; it is surprising that this was more severe than an analogous C2658A x G2663U mutation. A double transition mutation, C2658U x G2663A, which is not known to have occurred in nature, had no effect on the growth of cells or on the function of ribosomes. The lethal phenotype of transversion mutations in C2658 and G2663 appears to derive from a loss of the capacity of ribosomes to bind EF-G and by indirection the EF-Tu ternary complex., (Copyright 2000 Academic Press.)
- Published
- 2000
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13. The contribution of a zinc finger motif to the function of yeast ribosomal protein YL37a.
- Author
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Rivlin AA, Chan YL, and Wool IG
- Subjects
- Amino Acid Sequence, Binding Sites genetics, Cysteine genetics, Escherichia coli genetics, Fungal Proteins chemistry, Genes, Fungal, Genetic Complementation Test, Molecular Sequence Data, Mutation, Phenotype, Protein Binding, Recombinant Proteins chemistry, Recombinant Proteins genetics, Recombinant Proteins metabolism, Ribosomal Proteins chemistry, Zinc metabolism, Zinc Fingers genetics, Fungal Proteins genetics, Fungal Proteins metabolism, Ribosomal Proteins genetics, Ribosomal Proteins metabolism, Saccharomyces cerevisiae genetics, Saccharomyces cerevisiae metabolism
- Abstract
Eukaryotic ribosomes have a large number of proteins but the exact nature of their contribution to the structure and to the function of the particle is not known. Of the 78 proteins in yeast ribosomes, six have zinc finger motifs of the C2-C2 variety. Both genes encoding the essential yeast ribosomal protein YL37a, which has such a zinc finger motif, were disrupteXXPd. The double deletion, which is lethal, can be rescued with a plasmid-encoded copy of a YL37a gene. Mutations were constructed in a plasmid-encoded copy of YL37a; the mutations caused the cysteine residues in the motif (at positions 39, 42, 57 and 60) to be replaced, one at a time, with serine. The cysteine residue at position 39, the first of the four in the motif, is essential for the function of YL37a, since a C39S mutation did not complement the null phenotype. However, plasmids encoding variants with C42S, C57S, or C60S mutations in the zinc finger motif were able to rescue the null mutant. YL37a binds zinc, but none of the mutant proteins, C39S, C42S, C57S, or C60S, was able to bind the metal. Thus, all four cysteine residues are essential for the binding of zinc; only one, C39, is essential for the function of the ribosomal protein., (Copyright 1999 Academic Press.)
- Published
- 1999
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14. The two faces of the Escherichia coli 23 S rRNA sarcin/ricin domain: the structure at 1.11 A resolution.
- Author
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Correll CC, Wool IG, and Munishkin A
- Subjects
- Animals, Crystallography, X-Ray, Hydrogen Bonding, Models, Molecular, Nucleic Acid Conformation, Peptide Elongation Factors chemistry, RNA-Binding Proteins chemistry, Rats, Thallium chemistry, Water chemistry, Escherichia coli genetics, RNA, Ribosomal, 23S chemistry, Ricin chemistry
- Abstract
The sarcin/ricin domain of 23 S - 28 S ribosomal RNA is essential for protein synthesis because it forms a critical part of the binding site for elongation factors. A crystal structure of an RNA of 27 nucleotides that mimics the domain in Escherichia coli 23 S rRNA was determined at 1.11 A resolution. The domain folds into a hairpin distorted by four non-canonical base-pairs and one base triple. The fold is stabilized by cross-strand and intra-stand stacking; no intramolecular stabilizing metal ions are observed. This is the first structure to reveal in great detail the geometry and the hydration of two common motifs that are conserved in this rRNA domain, a GAGA tetraloop and a G-bulged cross-strand A stack. Differences in the region connecting these motifs to the stem in the E. coli and in the rat sarcin/ricin domains may contribute to the species-specific binding of elongation factors. Correlation of nucleotide protection data with the structure indicates that the domain has two surfaces. One surface is accessible, lies primarily in the major groove, and is likely to bind the elongation factors. The second lies primarily in the minor groove, and is likely to be buried in the ribosome. This minor groove surface includes the Watson-Crick faces of the cytosine bases in the unusual A2654.C2666 and U2653.C2667 water-mediated base-pairs., (Copyright 1999 Academic Press.)
- Published
- 1999
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15. The phenotype of mutations of G2655 in the sarcin/ricin domain of 23 S ribosomal RNA.
- Author
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Macbeth MR and Wool IG
- Subjects
- Base Sequence, Cell Division genetics, Molecular Sequence Data, Nucleic Acid Conformation, Peptide Elongation Factors genetics, Phenotype, Plasmids genetics, Polyribosomes genetics, Protein Synthesis Inhibitors pharmacology, Ricin pharmacology, rRNA Operon genetics, Escherichia coli genetics, Mutation genetics, RNA, Ribosomal, 23S genetics, Ricin genetics
- Abstract
The sarcin/ricin domain (SRD) in Escherichia coli 23 S rRNA forms a part of the site for the association of the elongation factors with the ribosome and hence is critical for the binding of aminoacyl-tRNA and for translocation. The domain is also the site of action of the eponymous toxins which catalyze covalent modification of single nucleotides that inactivate the ribosome. The conformation of the conserved guanosine at position 2655 is an especially prominent feature of the structure of the SRD: the nucleotide is bulged out of a helix and forms a base-triple with A2665 and U2656. G2655 in 23 S rRNA is protected from chemical modification when the elongation factors, EF-Tu and EF-G, are bound to ribosomes and the analog of G2655 in oligoribonucleotides is critical for recognition by the toxin sarcin and by EF-G. The contribution of G2655 to the function of the ribosome has been evaluated by constructing mutations in the nucleotide and determining the phenotype. Constitutive expression of a plasmid-encoded rrnB operon with a deletion of, or transversions in, G2655 is lethal to E. coli cells, whereas a defect in the growth of cells with a G2655A transition is observed only in competition with wild-type cells. The sedimentation profiles of ribosomes with mutations in G2655 are altered; most markedly by deletion or transversion of the nucleotide, less severely by transition to adenosine. Mutations of G2655 confer resistance to sarcin on ribosomes. Ribosomes with G2655Delta, G2655C, or G2655U mutations in 23 S rRNA are not active in protein synthesis, whereas those with the G2655A transition mutation suffer decreased activity., (Copyright 1999 Academic Press.)
- Published
- 1999
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16. Characterization of in vitro and in vivo mutations in non-conserved nucleotides in the ribosomal RNA recognition domain for the ribotoxins ricin and sarcin and the translation elongation factors.
- Author
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Macbeth MR and Wool IG
- Subjects
- Animals, Base Sequence, Binding Sites, Catalysis, Escherichia coli genetics, Molecular Sequence Data, Mutagenesis, Nucleic Acid Conformation, Nucleotides, Peptide Elongation Factor G, Peptide Elongation Factors metabolism, Phenotype, Plasmids, Protein Biosynthesis, RNA, Bacterial, RNA, Ribosomal, 23S, RNA, Ribosomal, 28S genetics, Rats, rRNA Operon, Endoribonucleases metabolism, Fungal Proteins, RNA, Ribosomal, 28S metabolism, Ricin metabolism
- Abstract
The sarcin/ricin domain in 23 S/28 S rRNA is crucial for ribosome function, since it constitutes at least part of the binding site for the elongation factors and hence is essential for binding aminoacyl-tRNA and for translocation. The domain is also the site of action of ricin and sarcin and analysis of the effect of mutations in the RNA on recognition by the cytotoxins has helped to define the structure and to understand the function of the region. We have constructed deletions, separately, of pairs of non-conserved, juxtaposed but non-hydrogen-bonded nucleotides that correspond to C4317 and C4331, and to U4316 and C4332, in an oligoribonucleotide that mimics the sarcin/ricin domain in rat 28 S rRNA. The deletions had no effect on the depurination of A4324 by ricin nor on the cleavage of the phosphodiester bond on the 3' side of G4325 by sarcin. However, simultaneous deletion of the four nucleotides decreased cleavage by sarcin but did not affect depurination by ricin. Removal of the non-canonical A4318.A4330 pair abolished recognition by both toxins. Deletion from oligoribonucleotides, that reproduce the sarcin/ricin domain of Escherichia coli 23 S rRNA, of U2653 and C2667 (equivalent to U4316, C4317 and C4331, C4332 in 28 S rRNA), or substitution of guanosine for U2653 (designed to form a Watson-Crick G2653.C2667 pair), reduced cleavage by sarcin whereas depurination by ricin was slightly increased. An increase in the stability of the mutant oligoribonucleotides may be the basis of the impairment in sarcin action. The tm for the wild-type RNA is 60 degreesC; for the double-deletion mutant U2653Delta/C2667Delta it is 65 degreesC; and for the U2653G transversion it is 69 degreesC. Expression of a mutant 23 S rRNA gene lacking U2653 and C2667 is lethal and a U2653G transversion mutation impairs growth. The mutant ribosomes are less active in protein synthesis than the wild-type and ribosomes with the U2653G mutation are resistant to sarcin. The binding of EF-G to oligoribonucleotides with a U2653/C2667 double deletion is reduced and an effect on the affinity of the factor for the sarcin/ricin domain may account in part for the decrease in ribosome efficiency. The results stress the potential importance in rRNA structure and function of non-conserved nucleotides, and suggest that the sarcin/ricin domain in ribosomes requires a region of structural flexibility for optimal efficiency., (Copyright 1999 Academic Press.)
- Published
- 1999
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17. Crystal structure of the ribosomal RNA domain essential for binding elongation factors.
- Author
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Correll CC, Munishkin A, Chan YL, Ren Z, Wool IG, and Steitz TA
- Subjects
- Animals, Binding Sites, Escherichia coli, Magnetic Resonance Spectroscopy, Molecular Sequence Data, Peptide Elongation Factors chemistry, Protein Biosynthesis, RNA, Ribosomal, 28S metabolism, Rats, Nucleic Acid Conformation, Peptide Elongation Factors metabolism, RNA, Ribosomal, 28S chemistry
- Abstract
The structure of a 29-nucleotide RNA containing the sarcin/ricin loop (SRL) of rat 28 S rRNA has been determined at 2.1 A resolution. Recognition of the SRL by elongation factors and by the ribotoxins, sarcin and ricin, requires a nearly universal dodecamer sequence that folds into a G-bulged cross-strand A stack and a GAGA tetraloop. The juxtaposition of these two motifs forms a distorted hairpin structure that allows direct recognition of bases in both grooves as well as recognition of nonhelical backbone geometry and two 5'-unstacked purines. Comparisons with other RNA crystal structures establish the cross-strand A stack and the GNRA tetraloop as defined and modular RNA structural elements. The conserved region at the top is connected to the base of the domain by a region presumed to be flexible because of the sparsity of stabilizing contacts. Although the conformation of the SRL RNA previously determined by NMR spectroscopy is similar to the structure determined by x-ray crystallography, significant differences are observed in the "flexible" region and to a lesser extent in the G-bulged cross-strand A stack.
- Published
- 1998
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18. The ribosome-in-pieces: binding of elongation factor EF-G to oligoribonucleotides that mimic the sarcin/ricin and thiostrepton domains of 23S ribosomal RNA.
- Author
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Munishkin A and Wool IG
- Subjects
- Base Sequence, Escherichia coli genetics, Escherichia coli metabolism, Molecular Sequence Data, Oligonucleotides chemistry, Oligonucleotides genetics, Oligonucleotides metabolism, Peptide Elongation Factor G, Protein Binding, RNA, Ribosomal, 23S metabolism, Ribosomal Proteins metabolism, Ribosomes genetics, Ricin chemistry, Thiostrepton chemistry, Thiostrepton metabolism, Peptide Elongation Factors metabolism, RNA, Ribosomal, 23S chemistry, Ribosomal Proteins chemistry, Ribosomes chemistry, Ribosomes metabolism
- Abstract
An oligoribonucleotide (a 27-mer) that mimics the sarcin/ricin (S/R) domain of Escherichia coli 23S rRNA binds elongation factor EF-G; the Kd is 6.9 microM, whereas for binding to ribosomes it is 0.7 microM. Binding saturates when EF-G and the S/R RNA are equimolar; at saturation 70% of the input RNA is in complexes with EF-G. Binding of EF-G to S/R RNA does not require GTP but is inhibited by GDP; the inhibition by GDP is overcome by GTP. The effects of mutations of the S/R domain nucleotides G2655, A2660, and G2661 suggest that EF-G recognizes the conformation of the RNA rather than the identity of the nucleotides. EF-G also binds to an oligoribonucleotide (an 84-mer) that has the thiostrepton region of 23S rRNA; however, EF-G binds independently to S/R and thiostrepton oligoribonucleotides.
- Published
- 1997
- Full Text
- View/download PDF
19. The primary structures of rat ribosomal proteins S3a (the V-Fos transformation effector) and of S3b.
- Author
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Chan YL, Olvera J, Paz V, and Wool IG
- Subjects
- Alternative Splicing, Amino Acid Sequence, Animals, Base Sequence, DNA, Complementary genetics, Gene Dosage, Genes, fos, Molecular Sequence Data, Plant Proteins chemistry, Plant Proteins genetics, RNA Processing, Post-Transcriptional, RNA, Messenger genetics, RNA, Messenger metabolism, Rats, Ribosomal Proteins genetics, Cell Cycle Proteins, Ribosomal Proteins chemistry
- Abstract
The amino acid sequence of the rat 40S ribosomal subunit protein S3a was deduced from the sequence of nucleotides in two recombinant cDNAs and confirmed by the determination of the NH2-terminal sequence by Edman degradation. Ribosomal protein S3a has 263 amino acids (the NH2-terminal methionine is removed after translation of the mRNA) and the molecular weight is 29,794. The protein designated S3b has the same amino acid sequence as S3a except that it lacks the carboxyl-terminal 12 residues. We are unable to determine whether there are separate genes for S3a and S3b, or whether there is a single gene and alternate splicing of the precursor to yield separate mRNAs for S3a and S3b, or whether there is a single gene and a single mRNA whose translation yields S3a which is converted by proteolysis, either physiological or fortuitous, to S3b. The mRNA for S3a is about 1000 nucleotides in length. Hybridization of cDNA to digests of nuclear DNA suggests that there are 8-13 copies of the S3a gene. Rat ribosomal protein S3a is identical to the product of the rat Fte-1 gene which encodes the V-Fos transformation effector; S3a is also related to the plant protein cyc07, which is encoded by a cell cycle S-phase specific gene.
- Published
- 1996
- Full Text
- View/download PDF
20. The primary structure of rat ribosomal protein L10: relationship to a Jun-binding protein and to a putative Wilms' tumor suppressor.
- Author
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Chan YL, Diaz JJ, Denoroy L, Madjar JJ, and Wool IG
- Subjects
- Amino Acid Sequence, Animals, Base Sequence, Carrier Proteins metabolism, Chickens, DNA, Complementary genetics, Humans, Liver metabolism, Molecular Sequence Data, Molecular Structure, Multigene Family, RNA, Messenger chemistry, RNA, Messenger genetics, RNA, Messenger metabolism, Rats, Ribosomal Protein L10, Species Specificity, Carrier Proteins chemistry, Carrier Proteins genetics, Genes, Wilms Tumor, Proto-Oncogene Proteins c-jun metabolism, Ribosomal Proteins chemistry, Ribosomal Proteins genetics
- Abstract
The amino acid sequence of the rat 60S ribosomal subunit protein L10 was deduced from the sequence of nucleotides in two recombinant cDNAs and confirmed by determination of the NH2-terminal amino acid sequence in the protein. Ribosomal protein L10 has 213 amino acids (the NH2-terminal methionine is removed after translation of the mRNA); the molecular weight is 24,456. Hybridization of the cDNA to digests of nuclear DNA suggests that there are 8 to 10 copies of the L10 gene. The mRNA for the protein is about 900 nucleotides in length. Rat L10 is related to ribosomal proteins from other eukaryotes. Ribosomal protein L10 is, in addition, the mammalian homolog of the chicken Jun-binding protein and is nearly identical to a putative Wilms' tumor suppressor. This is a presumptive example, of which there are many others, of an extraribosomal function of a ribosomal protein.
- Published
- 1996
- Full Text
- View/download PDF
21. The primary structure of rat ribosomal protein L6.
- Author
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Chan YL and Wool IG
- Subjects
- Amino Acid Sequence, Amino Acids analysis, Animals, Base Sequence, Cloning, Molecular, Conserved Sequence genetics, DNA, Complementary chemistry, Databases, Factual, Gene Dosage, Molecular Sequence Data, Molecular Weight, Open Reading Frames genetics, Organophosphorus Compounds metabolism, Peptide Fragments chemistry, RNA, Messenger chemistry, Rats, Ribosomal Proteins isolation & purification, Sequence Analysis, Software, Ribosomal Proteins chemistry
- Abstract
The amino acid sequence of the rat 60S ribosomal subunit protein L6 was deduced from the sequence of nucleotides in a recombinant cDNA and confirmed by determination of amino acid sequences in the protein. Ribosomal protein L6 has 296 amino acids (the NH2-terminal methionine is removed after translation of mRNA); the molecular weight is 33,417. Hybridization of the cDNA to digests of nuclear DNA suggests that there are 7 to 10 copies of the L6 gene. The mRNA for the protein is about 1,100 nucleotides in length. Rat L6 is related to ribosomal proteins from other eukaryotes and to histone H1.
- Published
- 1996
- Full Text
- View/download PDF
22. Dependence of depurination of oligoribonucleotides by ricin A-chain on divalent cations and chelating agents.
- Author
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Glück A and Wool IG
- Subjects
- Base Sequence, Chelating Agents chemistry, Chelating Agents metabolism, Molecular Sequence Data, Nucleic Acid Conformation, Oligoribonucleotides chemistry, Oligoribonucleotides metabolism, RNA, Ribosomal metabolism, Ricin metabolism
- Abstract
Ricin A-chain is a cytotoxic RNA N-glycosidase that inactivates eukaryotic ribosomes by depurinating the adenosine at position 4324 in 28S rRNA. The enzyme retains its specificity when a synthetic oligoribonucleotide (a 35-mer) that mimics the structure at the site of action is the substrate. However, covalent modification by ricin A-chain of the oligoribonucleotide but not of ribosomes, depends on the simultaneous presence of a divalent cation and a chelating agent.
- Published
- 1996
- Full Text
- View/download PDF
23. Extraribosomal functions of ribosomal proteins.
- Author
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Wool IG
- Subjects
- DNA-Binding Proteins, Eukaryotic Cells, Prokaryotic Cells, Ribosomes, Biological Evolution, Ribosomal Proteins genetics
- Abstract
The discovery of DNA-binding motifs in ribosomal proteins has led to the conjecture that the transition of the ribosome from an RNA to an RNP machine occurred by adding pre-existing proteins. Supportive, but circumstantial, evidence for the hypothesis is adduced from the finding that many ribosomal proteins have a second function apart from the particle. These extraribosomal functions are enumerated.
- Published
- 1996
24. The primary structure of rat ribosomal protein L10a.
- Author
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Olvera J and Wool IG
- Subjects
- Amino Acid Sequence, Animals, Base Sequence, Cloning, Molecular, DNA, Complementary, Humans, Liver Regeneration, Molecular Sequence Data, Oligonucleotide Probes, RNA, Messenger biosynthesis, Rats, Recombinant Proteins biosynthesis, Recombinant Proteins chemistry, Restriction Mapping, Ribosomal Proteins biosynthesis, Sequence Homology, Amino Acid, Software, Species Specificity, Liver metabolism, Ribosomal Proteins chemistry
- Abstract
The amino acid sequence of the rat 60S ribosomal subunit protein L1Oa was deduced from the sequence of nucleotides in a recombinant cDNA. Ribosomal protein L10a has 217 amino acids and has a molecular weight of 24,815. Hybridization of the cDNA to digests of nuclear DNA suggests that there are 7 to 1O copies of the L10a gene. The mRNA for the protein is about 760 nucleotides in length. Rat L1Oa is related to ribosomal proteins from other eukaryotes and from archaebacteria.
- Published
- 1996
- Full Text
- View/download PDF
25. Determination of the 28 S ribosomal RNA identity element (G4319) for alpha-sarcin and the relationship of recognition to the selection of the catalytic site.
- Author
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Glück A and Wool IG
- Subjects
- Base Sequence, Binding Sites genetics, Hydrolysis, Models, Molecular, Molecular Sequence Data, Mutagenesis, Site-Directed, Nucleic Acid Conformation, Protein Biosynthesis, RNA, Ribosomal, 28S chemistry, Ricin metabolism, Endoribonucleases, Fungal Proteins metabolism, RNA, Ribosomal, 28S genetics, RNA, Ribosomal, 28S metabolism
- Abstract
Ricin A-chin and alpha-sarcin are ribotoxins that inactivate eukaryotic ribosomes by modifying 28 S rRNA; ricin A-chain is an RNA N-glycosidase that depurinates the adenosine at position 4324 and alpha-sarcin is a ribonuclease that cleaves the phosphodiester bond on the 3' side of the adjacent guanosine (at position 4325). In cartoons of the secondary structure these two residues are seen to be embedded in a 17 base single-stranded loop over a seven base-pair helix. However, NMR spectroscopy of an oligoribonucleotide, a 29-mer that mimics the sarcin/ricin domain, indicates that the RNA has a compact conformation in which the guanosine at the position analogous to 4319 in 28 S rRNA is bulged out of what otherwise is an extended A-form helix. Since similar structural irregularities are used by proteins to bind to RNA, we have tested the effect of mutations of the bulged guanosine on recognition and covalent modification of the RNA by ricin A-chain and by alpha-sacrin. For the test a synthetic oligoribonucletide, a 35-mer, was used; the mutations were the deletion, the transition to adenosine, and the transversion to cytidine and uridine of the guanosine that is the analog of G4319. Each of the four mutations abolished cleavage og the RNA by alpha-sacrin, where depurination by ricin A-chain was little affected. Thus G4319 is an identity element for alpha-sacrin recognition. Analysis of the effect of alpha-sacrin on variant oligoribonucleotides in which additional bases were inserted between the identity element guanosine and the site of catalysis suggest that on binding to the RNA the toxin uses the guanosine for orientation and then cleaves at a fixed distance and at a fixed position in space.
- Published
- 1996
- Full Text
- View/download PDF
26. Systematic deletion analysis of ricin A-chain function. Single amino acid deletions.
- Author
-
Munishkin A and Wool IG
- Subjects
- Amino Acid Sequence, Catalysis, Kinetics, Models, Molecular, Molecular Sequence Data, Mutagenesis, N-Glycosyl Hydrolases isolation & purification, RNA, Ribosomal, 28S metabolism, Recombinant Proteins chemistry, Recombinant Proteins isolation & purification, Recombinant Proteins metabolism, Restriction Mapping, Ribosome Inactivating Proteins, Ricin chemistry, Ricin isolation & purification, N-Glycosyl Hydrolases chemistry, N-Glycosyl Hydrolases metabolism, Protein Structure, Secondary, Ricin metabolism, Sequence Deletion
- Abstract
The A-chain of ricin is a cytotoxic RNA N-glycosidase that inactivates ribosomes by depurination of the adenosine at position 4324 in 28 S rRNA. Of the 267 amino acids in the protein, 222 could be deleted, in one or another of 74 mutants, without the loss of the capacity to catalyze hydrolysis of a single specific nucleotide in rRNA (Morris, K. N., and Wool, I. G. (1992) Proc. Natl. Acad. Sci. U.S.A. 89, 4869-4873). The 45 amino acids that could not be omitted when the deletions were in sets of 20, 5, or 2 residues have now been deleted one at a time; 9 of these deletion mutants retained activity. A RNP-like structural motif in ricin A-chain that may mediate binding to ribosomal RNA has been identified.
- Published
- 1995
- Full Text
- View/download PDF
27. Structure and evolution of mammalian ribosomal proteins.
- Author
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Wool IG, Chan YL, and Glück A
- Subjects
- Amino Acid Sequence, Animals, Base Sequence, Genetic Code, Humans, Molecular Sequence Data, Molecular Structure, RNA, Nuclear genetics, Rats, Biological Evolution, RNA, Messenger genetics, Ribosomal Proteins chemistry
- Abstract
Mammalian (rat) ribosomes have 80 proteins; the sequence of amino acids in 75 have been determined. What has been learned of the structure of the rat ribosomal proteins is reviewed with particular attention to their evolution and to amino acid sequence motifs. The latter include: clusters of basic or acidic residues; sequence repeats or shared sequences; zinc finger domains; bZIP elements; and nuclear localization signals. The occurrence and the possible significance of phosphorylated residues and of ubiquitin extensions is noted. The characteristics of the mRNAs that encode the proteins are summarized. The relationship of the rat ribosomal proteins to the proteins in ribosomes from humans, yeast, archaebacteria, and Escherichia coli is collated.
- Published
- 1995
- Full Text
- View/download PDF
28. The carboxyl extensions of two rat ubiquitin fusion proteins are ribosomal proteins S27a and L40.
- Author
-
Chan YL, Suzuki K, and Wool IG
- Subjects
- Amino Acid Sequence, Animals, Base Sequence, Cell Nucleus metabolism, DNA metabolism, DNA, Complementary chemistry, DNA, Complementary metabolism, Liver metabolism, Molecular Sequence Data, Open Reading Frames, Rats, Recombinant Fusion Proteins biosynthesis, Ribosomal Proteins biosynthesis, Sequence Homology, Amino Acid, Ubiquitins biosynthesis, Zinc Fingers, Recombinant Fusion Proteins chemistry, Ribosomal Proteins chemistry, Ubiquitins chemistry
- Abstract
Two rat recombinant cDNAs were characterized; they encode fusion proteins that have at their NH2 terminus the conserved 76 amino acid ubiquitin and at their carboxyl terminus the extension ribosomal proteins S27a (80 amino acids and a molecular weight of 9,397) or L40 (52 amino acids and a molecular weight of 6,177). The fusion proteins are processed in a reticulocyte lysate to ubiquitin and either S27a or L40. Hybridization of cDNAs to digests of nuclear DNA suggests that there are 14 to 19 copies of the S27a, and 6 to 10 of the L40, genes. The mRNA for ubiquitin-S27a has about 700 nucleotides and ubiquitin-L40 about 650. Ribosomal proteins S27a and L40 contain zinc finger motifs of the C2-C2 variety. S27a and L40 are related to ribosomal proteins from other species.
- Published
- 1995
- Full Text
- View/download PDF
29. The primary structures of rat ribosomal proteins L4 and L41.
- Author
-
Chan YL, Olvera J, and Wool IG
- Subjects
- Amino Acid Sequence, Amino Acids analysis, Animals, Base Sequence, Chromatography, High Pressure Liquid, DNA, Complementary, Humans, Molecular Sequence Data, RNA, Messenger analysis, RNA, Messenger biosynthesis, Rats, Recombinant Proteins biosynthesis, Recombinant Proteins chemistry, Recombinant Proteins isolation & purification, Repetitive Sequences, Nucleic Acid, Ribosomal Proteins biosynthesis, Ribosomal Proteins isolation & purification, Ribosomal Proteins chemistry
- Abstract
The amino acid sequences of the rat 60S ribosomal subunit proteins L4 and L41 were deduced from the sequences of nucleotides in recombinant cDNAs. Ribosomal protein L4 has 421 amino acids; the molecular weight is 47,280. L41 is the smallest ribosomal protein; it has 25 amino acids and a molecular weight of 3,454. Hybridization of the cDNAs to digests of nuclear DNA suggests that there are 7 to 8 copies of the L4, and 9 to 12 of the L41, genes. The mRNA for L4 is about 1,500 nucleotides in length and that for L41 about 500 nucleotides. The 5' noncoding sequence of the L4 cDNA is exceptional in that it has, in addition to a short polypyrimidine sequence at the 5' end, a second stretch of 15 consecutive pyrimidines near the site of initiation of translation. The 3' noncoding sequence of the L41 mRNA is unusual in that it is at least 246 nucleotides long. Rat L4 and L41 are related to ribosomal proteins from other eukaryotes.
- Published
- 1995
- Full Text
- View/download PDF
30. The primary structures of rat ribosomal proteins: the characterization of the cDNAs for S21 and L39, corrections in the sequences of L7 and L18a, and the identification of L33.
- Author
-
Chan YL, Olvera J, and Wool IG
- Subjects
- Amino Acid Sequence, Animals, Base Sequence, DNA, Complementary, Molecular Sequence Data, RNA, Messenger genetics, Rats, Aniline Compounds, Ribosomal Proteins genetics
- Abstract
cDNAs for rat ribosomal proteins S21 and L39 were characterized. S21 has 83 amino acids and a molecular weight of 9,121; L39 has 50 amino acids and a molecular weight of 6,271. There are 8 to 9 copies of the S21, and 9 to 11 copies of the L39 genes. The mRNA for S21 is about 500 nucleotides in length and for L39 it is about 450. Rat S21 and L39 are related to ribosomal proteins from other species. Corrections have been made in the amino acid sequences of L7 and L18a. The rat ribosomal protein designated L33 from its coordinates on two-dimensional gels has been found, from its amino acid sequence, to be identical to S24.
- Published
- 1995
- Full Text
- View/download PDF
31. The primary structure of rat ribosomal protein L22.
- Author
-
Chan YL and Wool IG
- Subjects
- Amino Acid Sequence, Animals, Base Sequence, DNA, Complementary, Molecular Sequence Data, Rats, RNA-Binding Proteins genetics, Ribosomal Proteins genetics
- Abstract
The amino acid sequence of the rat 60S ribosomal subunit protein L22 was deduced from the sequence of nucleotides in two recombinant cDNAs. Ribosomal protein L22 has 128 amino acids; the molecular weight is 14,779. Hybridization of the cDNA to digests of nuclear DNA suggests that there are 6 to 9 copies of the L22 gene. The mRNA for the protein is about 600 nucleotides in length. Rat L22 is related to ribosomal proteins from other eukaryotes and is identical in amino acid sequence to human EAP, the EBER 1 (Epstein-Barr virus encoded RNA) associated protein.
- Published
- 1995
- Full Text
- View/download PDF
32. Analysis of the contribution of an amphiphilic alpha-helix to the structure and to the function of ricin A chain.
- Author
-
Morris KN and Wool IG
- Subjects
- Amino Acid Sequence, Models, Molecular, Molecular Sequence Data, N-Glycosyl Hydrolases genetics, Recombinant Proteins chemistry, Ribosome Inactivating Proteins, Ricin genetics, Sequence Analysis, Sequence Deletion, Structure-Activity Relationship, N-Glycosyl Hydrolases chemistry, Protein Structure, Secondary, Ricin chemistry
- Abstract
The A chain of ricin is a cytotoxic RNA N-glycosidase that inactivates eukaryotic ribosomes. The contribution of the amphiphilic helix D, which is distant from the active site, to the catalysis of the depurination of the adenosine at position 4324 in 28S rRNA has been examined by systematic deletion of amino acids. Two sets of consecutive two- or three-amino acid deletions of the 12 residues in helix D, a total of 20 mutants, were constructed. All 12 of the amino acids could be deleted in one mutant or another without loss of activity; however, mutations that disrupted the amphiphilicity of the helix led to inactivation of the enzyme. Thus, the minimum contribution of helix D to the structure of the ricin A chain is to provide hydrophobic and hydrophilic surfaces to shield helix E, which has the active-site residues; moreover, no amino acid side chain in helix D makes a specific contribution to the recognition of the RNA substrate or to catalysis; and, finally, phasing of the amino acid deletions can be important to the phenotype of mutants.
- Published
- 1994
- Full Text
- View/download PDF
33. The primary structure of rat ribosomal protein L24.
- Author
-
Chan YL, Olvera J, and Wool IG
- Subjects
- Amino Acid Sequence, Animals, Base Sequence, Codon, DNA Probes, DNA, Complementary chemistry, Liver chemistry, Molecular Sequence Data, Molecular Weight, Nucleic Acid Hybridization, RNA, Messenger chemistry, Rats, Ribosomal Proteins genetics, Sequence Alignment, Sequence Analysis, Sequence Homology, Software, Ribosomal Proteins chemistry
- Abstract
The amino acid sequence of the rat 60S ribosomal subunit protein L24 was deduced from the sequence of nucleotides in a recombinant cDNA. Ribosomal protein L24 has 157 amino acids; the molecular weight is 17,767. Hybridization of the cDNA to digests of nuclear DNA suggests that there are 9 to 11 copies of the L24 gene. The mRNA for the protein is about 650 nucleotides in length. Rat L24 is related to ribosomal proteins from other eukaryotes and from archaebacteria.
- Published
- 1994
- Full Text
- View/download PDF
34. The primary structure of rat ribosomal protein S23.
- Author
-
Kitaoka Y, Olvera J, and Wool IG
- Subjects
- Amino Acid Sequence, Animals, Base Sequence, DNA, Complementary, Humans, Molecular Sequence Data, Molecular Weight, Protein Biosynthesis, RNA, Messenger metabolism, Rats, Recombinant Proteins biosynthesis, Recombinant Proteins chemistry, Ribosomal Proteins biosynthesis, Saccharomyces cerevisiae metabolism, Sequence Homology, Amino Acid, Species Specificity, Tetrahymena thermophila metabolism, Transcription, Genetic, Ribosomal Proteins chemistry
- Abstract
The amino acid sequence of the rat 40S ribosomal subunit protein S23 was deduced from the sequence of nucleotides in a recombinant cDNA. Ribosomal protein S23 has 142 amino acids, the NH2-terminal methionine is removed after translation of the mRNA, and a molecular weight of 15,666. Hybridization of the cDNA to digests of nuclear DNA suggests that there are 6 to 13 copies of the S23 gene. The mRNA for the protein is about 650 nucleotides in length. Rat S23 is identical to a human ribosomal protein and is also related to Saccharomyces cerevisiae S28, to Tetrahymena thermophila S12, and to the prokaryotic S12 family of ribosomal proteins.
- Published
- 1994
- Full Text
- View/download PDF
35. The primary structure of rat ribosomal protein L13.
- Author
-
Olvera J and Wool IG
- Subjects
- Amino Acid Sequence, Animals, Base Sequence, Cloning, Molecular, DNA, Complementary genetics, Gene Expression, Genes, Molecular Sequence Data, RNA, Messenger genetics, Rats, Sequence Homology, Amino Acid, Ribosomal Proteins chemistry
- Abstract
The amino acid sequence of the rat 60S ribosomal subunit protein L13 was deduced from the sequence of nucleotides in two recombinant cDNAs. Ribosomal protein L13 has 210 amino acids, the NH2-terminal methionine is removed after translation of the mRNA and has a molecular weight of 24,094. Hybridization of the cDNA to digests of nuclear DNA suggests that there are 8 to 10 copies of the L13 gene. The mRNA for the protein is about 870 nucleotides in length. Rat L13 is related to ribosomal proteins from other eukaryotes.
- Published
- 1994
- Full Text
- View/download PDF
36. The primary structure of rat ribosomal protein L15.
- Author
-
Chang YL, Olvera J, and Wool IG
- Subjects
- Amino Acid Sequence, Animals, Base Sequence, Cloning, Molecular, DNA, Complementary, Gene Expression, Genes, Molecular Sequence Data, RNA, Messenger genetics, Rats, Sequence Alignment, Sequence Homology, Amino Acid, Ribosomal Proteins chemistry
- Abstract
The amino acid sequence of the rat 60S ribosomal subunit protein L15 was deduced from the sequence of nucleotides in two recombinant cDNAs. Ribosomal protein L15 has 203 amino acids, the NH2-terminal methionine is removed after translation of the mRNA, and has a molecular weight of 24,000. Hybridization of the cDNA to digests of nuclear DNA suggests that there are 13 to 15 copies of the L15 gene. The mRNA for the protein is about 850 nucleotides in length. Rat L15 is related to ribosomal proteins from other eukaryotes. Rat L15 has the hexapeptide, TYKFFE, that also occurs in the amyloidogenic glycoprotein A4 which is associated with Alzheimer's disease and Down's Syndrome.
- Published
- 1994
- Full Text
- View/download PDF
37. The primary structure of rat ribosomal protein S15a.
- Author
-
Chan YL, Olvera J, Paz V, and Wool IG
- Subjects
- Amino Acid Sequence, Animals, Base Sequence, Cloning, Molecular, Genes, Molecular Sequence Data, Rats, Sequence Alignment, Sequence Homology, Amino Acid, Ribosomal Proteins chemistry
- Abstract
The amino acid sequence of the rat 40S ribosomal subunit protein S15a was deduced from the sequence of nucleotides in two recombinant cDNAs. Ribosomal protein S15a has 129 amino acids, the NH2-terminal methionine is removed after translation of the mRNA and has a molecular weight of 14,698. Hybridization of the cDNA to digests of nuclear DNA suggests that there are 10 to 15 copies of the S15a gene. The mRNA for the protein is about 650 nucleotides in length. Rat S15a is related to ribosomal proteins from other eukaryotes, from eubacteria, and from archaebacteria.
- Published
- 1994
- Full Text
- View/download PDF
38. A leucine zipper-like motif and a basic region-leucine zipper-like element in rat ribosomal protein L13a. Identification of the tum- transplantation antigen P198.
- Author
-
Chan YL, Olvera J, Glück A, and Wool IG
- Subjects
- Amino Acid Sequence, Animals, Base Sequence, DNA, Complementary, Mitochondrial Proteins, Molecular Sequence Data, RNA, Messenger genetics, Rats, Ribosomal Proteins genetics, Sequence Homology, Amino Acid, Species Specificity, Histocompatibility Antigens analysis, Leucine Zippers, Ribosomal Proteins chemistry, Saccharomyces cerevisiae Proteins
- Abstract
The amino acid sequence of the rat 60 S ribosomal subunit protein L13a was deduced from the sequence of nucleotides in two recombinant cDNAs. Mature ribosomal protein L13a has 202 amino acids (the NH2-terminal methionine is removed after translation of the mRNA) and a M(r) of 23,330. Hybridization of the L13a cDNA to digests of nuclear DNA suggests that there are 9-11 copies of the L13a gene. The mRNA for the protein is approximately 800 nucleotides in length. Rat L13a is related to the Saccharomyces cerevisiae ribosomal proteins that have been provisionally designated rp22 and rp23 and to the eubacterial and archaebacterial family of L13 ribosomal proteins. The mouse tum- transplantation antigen P198 is a mutant of the mouse homolog of rat ribosomal protein L13a. Rat ribosomal protein L7 has, at its NH2 terminus, five tandem repeats of a similar sequence of 12 amino acids (Lin, A., Chan, Y. L., McNally, J., Peleg, D., Meyuhas, O., and Wool, I. G. (1987) J. Biol. Chem. 262, 12665-12671); L13a has, in its carboxyl-terminal region, amino acid sequences with significant identity to L7 repeats 1, 3, and 5. L13a also has a number of short amino acid sequences that are repeated, a leucine zipper-like motif at its NH2 terminus, and a potential basic region-leucine zipper element in its carboxyl-terminal region.
- Published
- 1994
39. The ribosomal RNA identity elements for ricin and for alpha-sarcin: mutations in the putative CG pair that closes a GAGA tetraloop.
- Author
-
Glück A, Endo Y, and Wool IG
- Subjects
- Base Sequence, Binding Sites, Hydrogen Bonding, In Vitro Techniques, Kinetics, Molecular Sequence Data, Nucleic Acid Conformation, Substrate Specificity, Endoribonucleases, Fungal Proteins chemistry, RNA, Ribosomal, 28S chemistry, Ricin chemistry
- Abstract
alpha-Sarcin is a ribonuclease that cleaves the phosphodiester bond on the 3' side of G4325 in 28S rRNA; ricin A-chain is a RNA N-glycosidase that depurinates the 5' adjacent A4324. These single covalent modifications inactivate the ribosome. An oligoribonucleotide that reproduces the structure of the sarcin/ricin domain in 28S rRNA was synthesized and mutations were constructed in the 5' C and the 3' G that surround a GAGA tetrad that has the sites of toxin action. Covalent modification of the RNA by ricin, but not by alpha-sarcin, requires a Watson-Crick pair to shut off a putative GAGA tetraloop. Either the recognition elements for the two toxins are different despite their catalyzing covalent modification of adjacent nucleotides in 28S rRNA or there are transitions in the conformation of the alpha-sarcin/ricin domain in 28S rRNA and one conformer is recognized by alpha-sarcin and the other by ricin A-chain.
- Published
- 1994
- Full Text
- View/download PDF
40. The conformation of the sarcin/ricin loop from 28S ribosomal RNA.
- Author
-
Szewczak AA, Moore PB, Chang YL, and Wool IG
- Subjects
- Animals, Base Sequence, Hydrogen Bonding, Magnetic Resonance Spectroscopy, Models, Molecular, Molecular Sequence Data, Nucleic Acid Conformation, Oligonucleotides chemistry, RNA, Ribosomal, 28S chemistry, Rats, Thermodynamics, Endoribonucleases, Fungal Proteins toxicity, RNA, Ribosomal, 28S ultrastructure, Ricin toxicity
- Abstract
The sarcin/ricin loop is a highly conserved sequence found in the RNA of all large ribosomal subunits. The cytotoxins alpha-sarcin and ricin both inactivate ribosomes by cleaving a single bond in that loop. Once it has been attacked, ribosomes no longer interact with elongation factors properly, and translation stops. We have determined the conformation of the sarcin/ricin loop by multinuclear NMR spectroscopy using E73, a 29-nucleotide RNA that has the sarcin/ricin loop sequence and that is sensitive to both toxins in vitro. The sarcin/ricin loop has a compact structure that contains several purine.purine base pairs, a GAGA tetraloop, and a bulged guanosine adjacent to a reverse Hoogsteen A.U pair. It is stabilized by an unusual set of cross-strand base-stacking interactions and imino proton to phosphate oxygen hydrogen bonds. In addition to having interesting structural features, this model explains many of the biochemical observations made about the loop's structure and its reactivity with cytotoxins, and it sheds light on the loop's interactions with elongation factors.
- Published
- 1993
- Full Text
- View/download PDF
41. The carboxyl extension of a ubiquitin-like protein is rat ribosomal protein S30.
- Author
-
Olvera J and Wool IG
- Subjects
- Amino Acid Sequence, Animals, Base Sequence, DNA, Molecular Sequence Data, Molecular Weight, Oligodeoxyribonucleotides, Open Reading Frames, Protein Biosynthesis, RNA, Messenger biosynthesis, Rats, Recombinant Fusion Proteins chemistry, Recombinant Proteins chemistry, Ribosomal Proteins chemistry, Sequence Homology, Amino Acid, Sequence Homology, Nucleic Acid, Transcription, Genetic, Ubiquitins chemistry, RNA, Messenger metabolism, Ribosomal Proteins genetics, Ubiquitins genetics
- Abstract
The amino acid sequence of the rat 40 S ribosomal subunit protein S30 was deduced from the sequence of nucleotides in a recombinant cDNA and confirmed by the determination of the 18 residues at the NH2 terminus of the protein. Unlike the majority of ribosomal proteins, which are unprocessed primary products of the translation of their mRNAs, S30 is formed by cleavage from a larger hybrid protein. The NH2-terminal polypeptide has 38% identity with ubiquitin and contains the characteristic carboxyl-terminal Gly-Gly dipeptide of this family of proteins. S30 has 59 amino acids and the molecular weight is 6,643; the ubiquitin-like sequence has 74 residues and the molecular weight is 7,634. The hybrid protein is encoded in each of the 8-10 members of the family of rat S30 genes; there is, however, only a single species of mRNA which contains the sequences for both proteins. The coding sequence of the hybrid protein occurs in the reverse polarity in the genome of the Finkel-Biskis-Reilly murine sarcoma virus.
- Published
- 1993
42. The primary structure of rat ribosomal protein S9.
- Author
-
Chan YL, Paz V, Olvera J, and Wool IG
- Subjects
- Amino Acid Sequence, Amino Acids analysis, Animals, Base Sequence, DNA, Molecular Sequence Data, RNA, Messenger genetics, Rats, Repetitive Sequences, Nucleic Acid, Ribosomal Protein S9, Ribosomal Proteins genetics, Sequence Homology, Amino Acid, Ribosomal Proteins chemistry
- Abstract
The amino acid sequence of the rat 40S ribosomal subunit protein S9 was deduced from the sequence of nucleotides in a recombinant cDNA. Ribosomal protein S9 has 193 amino acids, the NH2-terminal methionine is removed after translation of the mRNA, and has a molecular weight of 22,360. Hybridization of the cDNA to digests of nuclear DNA suggests that there are 14 to 16 copies of the S9 gene. The mRNA for the protein is about 1,000 nucleotides in length in part because of an especially long 5' noncoding region (103 nucleotides). Rat S9 is related to ribosomal proteins from other eukaryotes, Saccharomyces cerevisiae YS11 and Dictyostelium discoideum rp 1024, and to the eubacterial, archaebacterial, and chloroplast family of S4 ribosomal proteins. We have identified the product of the Trypanosoma brucei gene U as the homolog of rat ribosomal protein S9.
- Published
- 1993
- Full Text
- View/download PDF
43. The primary structure of L37--a rat ribosomal protein with a zinc finger-like motif.
- Author
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Chan YL, Paz V, Olvera J, and Wool IG
- Subjects
- Amino Acid Sequence, Animals, Base Sequence, DNA, Recombinant, Molecular Sequence Data, RNA, Messenger genetics, Rats, Ribosomal Proteins genetics, Sequence Homology, Amino Acid, Ribosomal Proteins chemistry, Saccharomyces cerevisiae Proteins, Zinc Fingers genetics
- Abstract
The amino acid sequence of the rat 60S ribosomal subunit protein L37 was deduced from the sequence of nucleotides in a recombinant cDNA. Ribosomal protein L37 has 96 amino acids, the NH2-terminal methionine is removed after translation of the mRNA, and has a molecular weight of 10,939. Ribosomal protein L37 has a single zinc finger-like motif of the C2-C2 type. Hybridization of the cDNA to digests of nuclear DNA suggests that there are 13 or 14 copies of the L37 gene. The mRNA for the protein is about 500 nucleotides in length. Rat L37 is related to Saccharomyces cerevisiae ribosomal protein YL35 and to Caenorhabditis elegans L37. We have identified in the data base a DNA sequence that encodes the chicken homolog of rat L37.
- Published
- 1993
- Full Text
- View/download PDF
44. The primary structure of rat ribosomal protein L36.
- Author
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Chan YL, Paz V, Olvera J, and Wool IG
- Subjects
- Amino Acid Sequence, Amino Acids analysis, Animals, Base Sequence, DNA, Molecular Sequence Data, Rats, Ribosomal Proteins genetics, Sequence Homology, Amino Acid, Species Specificity, Ribosomal Proteins chemistry
- Abstract
The amino acid sequence of the rat 60S ribosomal subunit protein L36 was deduced from the sequence of nucleotides in a recombinant cDNA. Ribosomal protein L36 has 104 amino acids, the NH2-terminal methionine is removed after translation of the mRNA and has a molecular weight of 12,128. Hybridization of the cDNA to digests of nuclear DNA suggests that there are 8 to 11 copies of the L36 gene. The mRNA for the protein is about 500 nucleotides in length. Rat L36 is related to yeast ribosomal protein YL39.
- Published
- 1993
- Full Text
- View/download PDF
45. Zinc finger-like motifs in rat ribosomal proteins S27 and S29.
- Author
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Chan YL, Suzuki K, Olvera J, and Wool IG
- Subjects
- Amino Acid Sequence, Amino Acids analysis, Animals, Base Sequence, DNA, Electrophoresis, Polyacrylamide Gel, Humans, Molecular Sequence Data, Protein Binding, RNA, Ribosomal metabolism, RNA-Binding Proteins, Rats, Ribosomal Proteins genetics, Ribosomal Proteins metabolism, Sequence Homology, Amino Acid, Species Specificity, Zinc analysis, Metalloproteins, Nuclear Proteins, Ribosomal Proteins chemistry, Zinc Fingers
- Abstract
The primary structures of the rat 40S ribosomal subunit proteins S27 and S29 were deduced from the sequences of nucleotides in recombinant cDNAs and confirmed by determination of amino acid sequences in the proteins. Ribosomal protein S27 has 83 amino acids and the molecular weight is 9,339. Hybridization of cDNA to digests of nuclear DNA suggests that there are 4-6 copies of the S27 gene; the mRNA for the protein is about 620 nucleotides in length. Ribosomal protein S29 has 55 amino acids and the molecular weight is 6,541. There are 14-17 copies of the S29 gene and its mRNA is about 500 nucleotides in length. Rat ribosomal protein S29 is related to several members of the archaebacterial and eubacterial S14 family of ribosomal proteins. S27 and S29 have zinc finger-like motifs as do other proteins from eukaryotic, archaebacterial, eubacterial, and mitochondrial ribosomes. Moreover, ribosomes and ribosomal subunits appear to contain zinc and iron as well.
- Published
- 1993
- Full Text
- View/download PDF
46. The primary structure of rat ribosomal protein L23a. The application of homology search to the identification of genes for mammalian and yeast ribosomal proteins and a correlation of rat and yeast ribosomal proteins.
- Author
-
Suzuki K and Wool IG
- Subjects
- Amino Acid Sequence, Aminohydrolases genetics, Animals, Candida albicans genetics, Cloning, Molecular, DNA genetics, DNA, Fungal genetics, Formate-Tetrahydrofolate Ligase genetics, Methylenetetrahydrofolate Dehydrogenase (NADP) genetics, Molecular Sequence Data, Multienzyme Complexes genetics, Rats, Saccharomyces cerevisiae genetics, Sequence Alignment, Sequence Homology, Amino Acid, Escherichia coli Proteins, Genes, Genes, Fungal, Ribosomal Proteins genetics
- Abstract
The amino acid sequence of the rat 60 S ribosomal subunit protein L23a was deduced from the sequence of nucleotides in a recombinant cDNA. Ribosomal protein L23a has 156 amino acids and a molecular weight of 17,684. Hybridization of the L23a cDNA to digests of nuclear DNA suggests that there are 18-20 copies of the L23a gene. The mRNA for the protein is about 600 nucleotides in length. Rat L23a is related to the yeast Saccharomyces cerevisiae L25, to the archaebacterial Methanococcus vannielii L23, to eubacterial Escherichia coli L23, and to other members of the L23 family of ribosomal proteins. A novel application of a routine homology search procedure was employed to identify a nucleotide sequence that could be used to design an oligodeoxynucleotide probe to screen a library for a cDNA that encodes rat L23a; this same procedure uncovered a number of previously unidentified genes for yeast ribosomal proteins in the GenBank DNA data base. In a correlation of rat and yeast ribosomal proteins 48 pairs are shown to be related.
- Published
- 1993
47. Ribosomal RNA identity elements for recognition by ricin and by alpha-sarcin: mutation in the putative CG pair that closes a GAGA tetraloop.
- Author
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Endo Y, Gluck A, and Wool IG
- Subjects
- Base Sequence, Binding Sites, Molecular Sequence Data, Nucleic Acid Conformation, RNA, Ribosomal, 28S chemistry, RNA, Ribosomal, 28S genetics, Endoribonucleases, Fungal Proteins metabolism, Mutation, RNA, Ribosomal, 28S metabolism, Ricin metabolism
- Abstract
Alpha sarcin is a ribonuclease that cleaves the phosphodiester bond on the 3' side of G4325 in 28S rRNA; ricin A-chain is a RNA N-glycosidase that depurinates the 5' adjacent A4324. These single covalent modifications inactivate the ribosomes. An oligoribonucleotide that reproduces the structure of the sarcin/ricin domain in 28S rRNA was synthesized and mutations were constructed in the 3'C and the 5'G that surround the GAGA tetrad that has the site of toxin action. Analysis indicates that catalysis by ricin requires a Watson-Crick pair to shut off a putative GAGA tetraloop, whereas, alpha sarcin does not. One interpretation is that there are alternate conformations of the sarcin/ricin domain in 28S rRNA and that one of the conformers is recognized by sarcin and the other by ricin A-chain. This switch in the structure could underlie the translocation of peptidyl-tRNA from the A to the P site and the vectoral displacement of mRNA one codon during elongation.
- Published
- 1993
48. The primary structure of rat ribosomal protein S5. A ribosomal protein present in the rat genome in a single copy.
- Author
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Kuwano Y, Olvera J, and Wool IG
- Subjects
- Amino Acid Sequence, Amino Acids analysis, Animals, Base Sequence, DNA genetics, DNA, Recombinant metabolism, Electrophoresis, Polyacrylamide Gel, Genome, Molecular Sequence Data, Protein Biosynthesis, Pseudogenes, RNA Precursors metabolism, RNA Processing, Post-Transcriptional, RNA, Messenger genetics, RNA, Messenger metabolism, Rats, Ribosomal Proteins biosynthesis, Ribosomal Proteins isolation & purification, Sequence Homology, Amino Acid, Transcription, Genetic, Ribosomal Proteins genetics
- Abstract
The amino acid sequence of the rat 40 S ribosomal subunit protein S5 was deduced from the sequence of nucleotides in a recombinant cDNA and confirmed by the determination, directly from the protein, of 17 residues near the NH2 terminus. S5 has 204 amino acids; the molecular weight is 22,863. The protein designated S5a has the same amino acid sequence as S5 except that it lacks the NH2-terminal 5 residues. It is not known whether the conversion of a portion of S5 to S5a is physiological or fortuitous. The mRNA for S5 has about 820 nucleotides. Hybridization of the S5 cDNA to digests of nuclear DNA indicates that the rat genome has only a single copy of the gene; this is in distinction to the mouse and human genomes which have three to six copies of the S5 gene. Rat ribosomal protein S5 is related to the eubacteria, the arachaebacteria, and the chloroplast family of S7 ribosomal proteins. There is a peptide of 16 residues at the carboxyl terminus of S5 that is highly conserved in 18 species spanning the three kingdoms and chloroplasts.
- Published
- 1992
49. The primary structure of rat ribosomal protein L3.
- Author
-
Kuwano Y and Wool IG
- Subjects
- Amino Acid Sequence, Animals, Base Sequence, DNA chemistry, DNA, Recombinant chemistry, Molecular Sequence Data, Molecular Weight, Nucleic Acid Hybridization, RNA, Messenger genetics, Rats, Ribosomal Protein L3, Ribosomal Proteins genetics, Sequence Homology, Nucleic Acid, Software, Ribosomal Proteins chemistry
- Abstract
The amino acid sequence of the rat 60S ribosomal subunit protein L3 was deduced from the sequence of nucleotides in a recombinant cDNA. Ribosomal protein L3 has 403 amino acids and has a molecular weight of 46,106. Hybridization of the cDNA to digests of nuclear DNA suggests that there are 7 to 9 copies of the L3 gene. The mRNA for the protein is about 1,400 nucleotides in length. Rat L3 is homologous to ribosomal proteins from other eukaryotes and to proteins from eubacterial, archaebacterial, and chloroplast ribosomes.
- Published
- 1992
- Full Text
- View/download PDF
50. The primary structure of rat ribosomal protein S25.
- Author
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Chan YL and Wool IG
- Subjects
- Amino Acid Sequence, Animals, Base Sequence, DNA genetics, Gene Library, Humans, Liver physiology, Molecular Sequence Data, Molecular Weight, Poly A genetics, RNA, Messenger genetics, Rats, Recombinant Proteins chemistry, Ribosomal Proteins chemistry, Sequence Homology, Nucleic Acid, Ribosomal Proteins genetics
- Abstract
The amino acid sequence of the rat 40S ribosomal subunit protein S25 was deduced from the sequence of nucleotides in a recombinant cDNA. Ribosomal protein S25 has 125 amino acids and has a molecular weight of 13,733. Hybridization of the cDNA to digests of nuclear DNA suggests that there are 19 to 22 copies of the S25 gene. The mRNA for the protein is about 550 nucleotides in length. Rat S25 is homologous to ribosomal proteins from other eukaryotes (human and yeast).
- Published
- 1992
- Full Text
- View/download PDF
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