25 results on '"Tinoco I"'
Search Results
2. The structure of the L3 loop from the hepatitis delta virus ribozyme: a syn cytidine.
- Author
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Lynch, S R and Tinoco, I
- Abstract
The structure of the L3 central hairpin loop isolated from the antigenomic sequence of the hepatitis delta virus ribozyme with the P2 and P3 stems from the ribozyme stacked on top of the loop has been determined by NMR spectroscopy. The 26 nt stem-loop structure contains nine base pairs and a 7 nt loop (5'-UCCUCGC-3'). This hairpin loop is critical for efficient catalysis in the intact ribozyme. The structure was determined using homonuclear and heteronuclear NMR techniques on non-labeled and15N-labeled RNA oligonucleotides. The overall root mean square deviation for the structure was 1.15 A (+/- 0.28 A) for the loop and the closing C.G base pair and 0.90 A (+/- 0.18 A) for the loop and the closing C.G base pair but without the lone purine in the loop, which is not well defined in the structure. The structure indicates a U.C base pair between the nucleotides on the 5'- and 3'-ends of the loop. This base pair is formed with a single hydrogen bond involving the cytosine exocyclic amino proton and the carbonyl O4 of the uracil. The most unexpected finding in the loop is a syn cytidine. While not unprecedented, syn pyrimidines are highly unusual. This one can be confidently established by intranucleotide distances between the ribose and the base determined by NMR spectroscopy. A similar study of the structure of this loop showed a somewhat different three-dimensional structure. A discussion of differences in the two structures, as well as possible sites of interaction with the cleavage site, will be presented.
- Published
- 1998
- Full Text
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3. Circular dichroism calculations for polyinosinic acid in proposed multi-stranded geometries.
- Author
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Cech, C. L., primary and Tinoco, I., additional
- Published
- 1976
- Full Text
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4. The kinetics of binding of U-U-C-A to a dodecanucleotide anticodon fragment from yeast tRNAPhe
- Author
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Yoon, K., primary, Turner, D. H., additional, Tinoco, I., additional, von der Haar, F., additional, and Cramer, F., additional
- Published
- 1976
- Full Text
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5. EF-G catalyzed translocation dynamics in the presence of ribosomal frameshifting stimulatory signals.
- Author
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Kim HK and Tinoco I Jr
- Subjects
- Bacterial Proteins genetics, Biocatalysis, Codon, DNA Polymerase III genetics, Fluorescence Resonance Energy Transfer, Mutation, Peptide Chain Elongation, Translational, RNA, Messenger chemistry, RNA, Transfer, Lys metabolism, Frameshifting, Ribosomal, Peptide Elongation Factor G metabolism
- Abstract
Programmed -1 ribosomal frameshifting (-1PRF) is tightly regulated by messenger RNA (mRNA) sequences and structures in expressing two or more proteins with precise ratios from a single mRNA. Using single-molecule fluorescence resonance energy transfer (smFRET) between (Cy5)EF-G and (Cy3)tRNALys, we studied the translational elongation dynamics of -1PRF in the Escherichia coli dnaX gene, which contains three frameshifting signals: a slippery sequence (A AAA AAG), a Shine-Dalgarno (SD) sequence and a downstream hairpin. The frameshift promoting signals mostly impair the EF-G-catalyzed translocation step of the two tRNALys and the slippery codons from the A- and P- sites. The hairpin acts as a road block slowing the translocation rate. The upstream SD sequence together with the hairpin promotes dissociation of futile EF-G and thus causes multiple EF-G driven translocation attempts. A slippery sequence also helps dissociation of the EF-G by providing alternative base-pairing options. These results indicate that frameshifting takes place during the repetitive ribosomal conformational changes associated with EF-G dissociation upon unsuccessful translocation attempts of the second slippage codon from the A- to the P- sites., (© The Author(s) 2016. Published by Oxford University Press on behalf of Nucleic Acids Research.)
- Published
- 2017
- Full Text
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6. A mutant RNA pseudoknot that promotes ribosomal frameshifting in mouse mammary tumor virus.
- Author
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Kang H and Tinoco I Jr
- Subjects
- Adenine, Animals, Base Composition, Base Sequence, Computer Simulation, Guanine, Magnetic Resonance Spectroscopy, Mice, Models, Molecular, Molecular Sequence Data, Point Mutation, Thermodynamics, Uracil, Frameshifting, Ribosomal, Mammary Tumor Virus, Mouse genetics, Nucleic Acid Conformation, RNA, Viral chemistry, RNA, Viral genetics
- Abstract
A single A-->G mutation that changes a potential A.U base pair to a G.U pair at the junction of the stems and loops of a non-frameshifting pseudoknot dramatically increases its frameshifting efficiency in mouse mammary tumor virus. The structure of the non-frameshifting pseudoknot APK has been found to be very different from that of pseudoknots that cause efficient frameshifting [Kang,H., Hines,J.V. and Tinoco,I. (1995) J. Mol. Biol. , 259, 135-147]. The 3-dimensional structure of the mutant pseudoknot was determined by restrained molecular dynamics based on NMR-derived interproton distance and torsion angle constraints. One striking feature of the mutant pseudoknot compared with the parent pseudoknot is that a G.U base pair forms at the top of stem 2, thus leaving only 1 nt at the junction of the two stems. The conformation is very different from that of the previously determined non-frameshifting parent pseudoknot, which lacks the A.U base pair at the top of the stem and has 2 nt between the stems. However, the conformation is quite similar to that of efficient frameshifting pseudoknots whose structures were previously determined by NMR. A single adenylate residue intervenes between the two stems and interrupts their coaxial stacking. This unpaired nucleotide produces a bent structure. The structural similarity among the efficient frameshifting pseudoknots indicates that a specific conformation is required for ribosomal frameshifting, further implying a specific interaction of the pseudoknot with the ribosome.
- Published
- 1997
- Full Text
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7. Synthesis and NMR of RNA with selective isotopic enrichment in the bases.
- Author
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SantaLucia J Jr, Shen LX, Cai Z, Lewis H, and Tinoco I Jr
- Subjects
- Adenine chemical synthesis, Cytidine chemical synthesis, Guanosine chemical synthesis, Magnetic Resonance Spectroscopy, RNA chemistry, Uracil chemical synthesis, Uridine chemical synthesis, Purine Nucleosides chemical synthesis, Pyrimidine Nucleosides chemical synthesis, RNA chemical synthesis
- Abstract
Efficient syntheses of pyrimidine and purine nucleosides and nucleotides with selective 13C enrichment in the base moieties are described. Uridine and cytidine are labeled at position C6 and adenosine and guanosine are labeled at position C8. The selectively labeled nucleosides were converted to nucleoside triphosphates and used with in vitro transcription to synthesize labeled RNA. Isotope-edited 12C and 13C sub-spectra of a omega 1-1/2-X-filtered NOESY experiment are demonstrated to be useful for making resonance assignments and for deriving structural information in large (> 20 nt) RNA molecules. The labeled RNAs also allow heteronuclear J-couplings and relaxation parameters to be measured without complications from 13C-13C J-couplings.
- Published
- 1995
- Full Text
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8. Use of ultra stable UNCG tetraloop hairpins to fold RNA structures: thermodynamic and spectroscopic applications.
- Author
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Molinaro M and Tinoco I Jr
- Subjects
- Base Sequence, Magnetic Resonance Spectroscopy, Molecular Sequence Data, Nucleic Acid Denaturation, Oligoribonucleotides chemical synthesis, Oligoribonucleotides chemistry, Thermodynamics, Nucleic Acid Conformation, RNA chemistry
- Abstract
RNA molecules of > 20 nucleotides have been the focus of numerous recent NMR structural studies. Several investigators have used the UNCG family of hairpins to ensure proper folding. We show that th UUCG hairpin has a minimum requirement of a two base-pair stem. Hairpins with a CG loop closing base pair and an initial 5'CG or 5'GC base pair have a melting temperature approximately 55 degrees C in 10 mM sodium phosphate. The high stability of even such small hairpins suggests that the hairpin can serve as a nucleation site for folding. For high resolution NMR work, the UNCG loop family (UACG in particular) provides excellent spectroscopic markers in one-dimensional exchangeable spectra, in two-dimensional COSY spectra and in NOESY spectra that clearly define it as forming a hairpin. This allows straightforward initiation of chemical shift assignments.
- Published
- 1995
- Full Text
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9. The solution structure of a d[C(TTCG)G] DNA hairpin and comparison to the unusually stable RNA analogue.
- Author
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James JK and Tinoco I Jr
- Subjects
- Magnetic Resonance Spectroscopy, Models, Molecular, Solutions, DNA chemistry, Nucleic Acid Conformation, RNA chemistry
- Abstract
The solution structure of the DNA analogue of the unusually stable r[C(UUCG)G] RNA hairpin, 5'-d[GGA-C(TTCG)GTCC]-3', has been determined by NMR spectroscopy, and its structure has been compared to that of the RNA molecule. The RNA molecule is compact and rigid with a highly structured loop. However, the DNA molecule is much less structured. The DNA hairpin contains a B-form stem of four base pairs. The terminal base pair frays, and the 3'-terminal nucleotides, C11 and C12, are in equilibrium between 2'-endo and 3'-endo conformations. Unlike the RNA loop, the DNA loop contains no syn nucleotides, and there is no evidence for base-base or base-phosphate hydrogen bonding in the loop. The loop is flexible, and reveals no specific internucleotide interactions.
- Published
- 1993
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10. Structure of a small RNA hairpin.
- Author
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Davis PW, Thurmes W, and Tinoco I Jr
- Subjects
- Base Sequence, Hydrogen Bonding, Magnetic Resonance Spectroscopy, Models, Molecular, Molecular Sequence Data, Thermodynamics, Nucleic Acid Conformation, RNA chemistry
- Abstract
The hairpin stem-loop form of the RNA oligonucleotide rCGC(UUU)GCG has been studied by NMR spectroscopy. In 10 mM phosphate buffer this RNA molecule forms a unimolecular hairpin with a stem of three base pairs and a loop of three uridines, as judged by both NMR and UV absorbance melting behavior. Distance and torsion angle restraints were determined using homonuclear proton-proton and heteronuclear proton-phosphorus 2-D NMR. These values were used in restrained molecular dynamics to determine the structure of the hairpin. The stem has characteristics of A-form geometry, although distortion from A-form occurs in the 3'-side of the stem, presumably to aid in accommodating the small loop. The loop nucleotides adopt C2'-endo conformations. NOE's strongly suggest stacking of the uracils with the stem, especially the first uracil on the 5'-side of the loop. The reversal of the chain direction in the loop seems to occur between U5 and U6. Loop structures produced by molecular dynamics simulations had a wide range of conformations and did not show stacking of the uracils. A flexible loop with significant dynamics is consistent with all the data.
- Published
- 1993
- Full Text
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11. Thermodynamic parameters for loop formation in RNA and DNA hairpin tetraloops.
- Author
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Antao VP and Tinoco I Jr
- Subjects
- Base Sequence, Molecular Sequence Data, Temperature, Thermodynamics, DNA chemistry, Nucleic Acid Conformation, Oligodeoxyribonucleotides chemistry, Oligoribonucleotides chemistry, RNA chemistry
- Abstract
We determined the melting temperatures (Tm) and thermodynamic parameters of 15 RNA and 19 DNA hairpins at 1 M NaCl, 0.01 M sodium phosphate, 0.1 mM EDTA, at pH 7. All these hairpins have loops of four bases, the most common loop size in 16S and 23S ribosomal RNAs. The RNA hairpins varied in loop sequence, loop-closing base pair (A.U, C.G, or G.C), base sequence of the stem, and stem size (four or five base pairs). The DNA hairpins varied in loop sequence, loop-closing base pair (C.G, or G.C), and base sequence of the four base-pair stem. Thermodynamic properties of a hairpin may be represented by nearest-neighbor interactions of the stem plus contributions from the loop. Thus, we obtained thermodynamic parameters for the formation of RNA and DNA tetraloops. For the tetraloops we studied, a free energy of loop formation (at 37 degrees C) of about +3 kcal/mol is most common for either RNA or DNA. There are extra stable loops with delta G degrees 37 near +1 kcal/mol, but the sequences are not necessarily the same for RNA and DNA. The closing base pair is also important; changing from C.G to G.C lowered the stability of several tetraloops in both RNA and DNA. These values will be useful in predicting RNA and DNA secondary structures.
- Published
- 1992
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12. Poly(rA) binds poly(rG).poly(rC) to form a triple helix.
- Author
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Chastain M and Tinoco I Jr
- Subjects
- Circular Dichroism, Macromolecular Substances, Nucleic Acid Conformation, Poly A chemistry, Poly C chemistry, Poly G chemistry, Spectrophotometry, Temperature, Poly A metabolism, Poly C metabolism, Poly G metabolism
- Abstract
Poly(rA) binds poly(rG).poly(rC) to form a triple helix. Evidence for this structure includes ultraviolet absorbance mixing curves and melting curves, and circular dichroism spectroscopy. The formation of the triple helix depends on the length of the poly(rC) strand. Triple helix forms when the average length is around 100 nucleotides but does not form when the average length is about 500 nucleotides.
- Published
- 1992
- Full Text
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13. A thermodynamic study of unusually stable RNA and DNA hairpins.
- Author
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Antao VP, Lai SY, and Tinoco I Jr
- Subjects
- Base Sequence, Consensus Sequence genetics, Molecular Sequence Data, Temperature, DNA chemistry, Nucleic Acid Conformation, Oligonucleotides chemistry, RNA chemistry
- Abstract
About 70% of the RNA tetra-loop sequences identified in ribosomal RNAs from different organisms fall into either (UNCG) or (GNRA) families (where N = A, C, G, or U; and R = A or G). RNA hairpins with these loop sequences form unusually stable tetra-loop structures. We have studied the RNA hairpin GGAC(UUCG)GUCC and several sequence variants to determine the effect of changing the loop sequence and the loop-closing base pair on the thermodynamic stability of (UNCG) tetra-loops. The hairpin GGAG(CUUG)CUCC with the conserved loop G(CUUG)C was also unusually stable. We have determined melting temperatures (Tm), and obtained thermodynamic parameters for DNA hairpins with sequences analogous to stable RNA hairpins with (UNCG), C(GNRA)G, C(GAUA)G, and G(CUUG)C loops. DNA hairpins with (TTCG), (dUdUCG), and related sequences in the loop, unlike their RNA counterparts, did not form unusually stable hairpins. However, DNA hairpins with the consensus loop sequence C(GNRA)G were very stable compared to hairpins with C(TTTT)G or C(AAAA)G loops. The C(GATA)G and G(CTTG)C loops were also extra stable. The relative stabilities of the unusually stable DNA hairpins are similar to those observed for their RNA analogs.
- Published
- 1991
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14. A proton NMR study of a DNA dumb-bell structure with hairpin loops of only two nucleotides: d(CACGTGTGTGCGTGCA).
- Author
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Rinkel LJ and Tinoco I Jr
- Subjects
- Base Sequence, Deuterium, Magnetic Resonance Spectroscopy, Molecular Sequence Data, Temperature, Water, DNA chemistry, Nucleic Acid Conformation, Polynucleotides chemistry
- Abstract
One- and two-dimensional nuclear magnetic resonance (NMR) experiments were used to study the conformation of the DNA hexadecanucleotide d(CACGTGTGTGCGTGCA) in aqueous solution. NMR spectra were recorded for the compound in D2O and in H2O/D2O (90/10) over the temperature range 1 degree C-60 degrees C. Assignments of imino proton resonances and of non-exchangeable proton resonances (except for some H4', H5' and H5" resonances) are given. The 1H-NMR spectra indicate that below about 20 degrees C, the compound exists as a single monomolecular species. Between 20 degrees C and 55 degrees C the oligonucleotide occurs as a mixture of structures in fast exchange on the NMR time scale, except for the temperature region 30 degrees - 34 degrees C, where substantial line broadening indicates intermediate exchange; above 60 degrees C the single strand predominates. The imino proton spectra, chemical shift values, and scalar coupling and NOE data reveal that the monomeric form, which is exclusively present below 20 degrees C, consists of a structure with a B-DNA double helix region of six base pairs, both ends of which are closed by hairpin loops of only two nucleotides, giving the molecule a dumbbell-like structure: [sequence: see text].
- Published
- 1991
- Full Text
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15. Base-base mismatches. Thermodynamics of double helix formation for dCA3XA3G + dCT3YT3G (X, Y = A,C,G,T).
- Author
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Aboul-ela F, Koh D, Tinoco I Jr, and Martin FH
- Subjects
- Base Composition, Base Sequence, Hydrogen Bonding, DNA, Nucleic Acid Conformation, Thermodynamics
- Abstract
Thermodynamic parameters for double strand formation have been measured for the sixteen double helices of the sequence dCA3XA3G.dCT3YT3G, with each of the bases A, C, G and T at the positions labelled X and Y. The results are analyzed in terms of nearest-neighbors and are compared with thermodynamic parameters for RNA secondary structure. At room temperature the sequence (Formula: see text) is more stable than (Formula: see text) and is similar in stability to (Formula: see text) and (Formula: see text) are least stable. At higher temperatures the sequences containing a G.C base pair become more stable than those containing only A.T. All molecules containing mismatches are destabilized with respect to those with only Watson-Crick pairing, but there is a wide range of destabilization. At room temperature the most stable mismatches are those containing guanine (G.T, G.G, G.A); the least stable contain cytosine (C.A, C.C). At higher temperatures pyrimidine-pyrimidine mismatches become the least stable.
- Published
- 1985
- Full Text
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16. A dynamic programming algorithm for finding alternative RNA secondary structures.
- Author
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Williams AL Jr and Tinoco I Jr
- Subjects
- Animals, DNA Restriction Enzymes metabolism, Deoxyribonuclease BamHI, Ribonuclease T1 metabolism, Tetrahymena genetics, Computers, Nucleic Acid Conformation, RNA, Ribosomal analysis, Software
- Abstract
Dynamic programming algorithms that predict RNA secondary structure by minimizing the free energy have had one important limitation. They were able to predict only one optimal structure. Given the uncertainties of the thermodynamic data and the effects of proteins and other environmental factors on structure, the optimal structure predicted by these methods may not have biological significance. We present a dynamic programming algorithm that can determine optimal and suboptimal secondary structures for an RNA. The power and utility of the method is demonstrated in the folding of the intervening sequence of the rRNA of Tetrahymena. By first identifying the major secondary structures corresponding to the lowest free energy minima, a secondary structure of possible biological significance is derived.
- Published
- 1986
- Full Text
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17. Small changes in free energy assignments for unpaired bases do not affect predicted secondary structures in single stranded RNA.
- Author
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Nussinov R, Tinoco I Jr, and Jacobson AB
- Subjects
- Base Composition, Computers, Methods, Thermodynamics, Base Sequence, Nucleic Acid Conformation, RNA, Messenger
- Abstract
We present extensive calculations of the secondary structure of mRNA which point to its insensitivity to small changes in the free energy assignments of single stranded regions. Truncating the free energies of hairpin loops, bulges, internal loops and multibranched junctions to two significant digits yields structures nearly identical to those generated using three digit values. The results show that one can safely use truncated values in RNA folding calculations. The implementation of these results enabled us to carry out secondary structure calculations on 2600 nucleotides in a single computer run.
- Published
- 1982
- Full Text
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18. DNA-RNA hybrid duplexes containing oligo(dA:rU) sequences are exceptionally unstable and may facilitate termination of transcription.
- Author
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Martin FH and Tinoco I Jr
- Subjects
- Base Sequence, Deoxyadenosines, Hot Temperature, Nucleic Acid Conformation, Oligonucleotides, Uracil, DNA metabolism, Nucleic Acid Hybridization, RNA metabolism, Transcription, Genetic
- Abstract
A DNA-RNA hybrid oligonucleotide duplex, dC(pA) 5pG:rC(pU)5pG, which contains a (dA:rU) 5 sequence, is at least 200 times less stable at room temperature than the corresponding duplex containing an (rA:dT) 5 sequence, rC(pA)5pG:dC(pT)5pG. This result provides an explanation for the finding that most primary RNA transcripts terminate in several consecutive rU residues, but not rA residues. It strongly supports the idea that instability of the DNA-RNA hybrid at the growing point of transcription plays a role in termination of transcription.
- Published
- 1980
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19. The tetraribonucleotide rCpGpCpG forms a left-handed Z-RNA double-helix.
- Author
-
Davis PW, Hall K, Cruz P, Tinoco I Jr, and Neilson T
- Subjects
- Base Sequence, Chlorates, Circular Dichroism, Magnetic Resonance Spectroscopy, Spectrophotometry, Ultraviolet, Nucleic Acid Conformation, Oligoribonucleotides, RNA, Double-Stranded
- Abstract
NMR and circular dichroism studies show that the RNA tetranucleotide rCpGpCpG can form a Z-RNA left-handed double-helix. In 1.0 M NaClO4, circular dichroism measurements indicate that the tetramer is in the A-form. In 6.0 M NaClO4, there is a characteristic change in the circular dichroism, indicating that the tetramer adopts a left-handed Z-form. This conformation is verified by phosphorus and proton NMR studies. The 31P spectrum shows a large downfield shift in one of the resonances upon an increase in salt concentration. Proton nuclear Overhauser effect (NOE) experiments indicate that the guanosines are in the syn conformation. These results are consistent with the formation of a Z-form double-helix.
- Published
- 1986
- Full Text
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20. Base pairing involving deoxyinosine: implications for probe design.
- Author
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Martin FH, Castro MM, Aboul-ela F, and Tinoco I Jr
- Subjects
- Hydrogen Bonding, Oligodeoxyribonucleotides chemical synthesis, Structure-Activity Relationship, Thermodynamics, Inosine analogs & derivatives
- Abstract
The thermal stability of oligodeoxyribonucleotide duplexes containing deoxyinosine (I) residues matched with each of the four normal DNA bases were determined by optical melting techniques. The duplexes containing at least one I were obtained by mixing equimolar amounts of an oligonucleotide of sequence dCA3XA3G with one of sequence dCT3YT3G where X and Y were A, C, G, T, or I. Comparison of optical melting curves yielded relative stabilities for the I-containing standard base pairs in an otherwise identical base-pair sequence. I:C pairs were found to be less stable than A:T pairs in these duplexes. Large neighboring-base effects upon stability were observed. For example, when (X,Y) = (I,A), the duplex is eight-fold more stable than when (X,Y) = (A,I). Independent of sequence effects the order of stabilities is: I:C greater than I:A greater than I:T congruent to I:G. This order differs from that of deoxyguanosine which pairs less strongly with dA; otherwise each deoxyinosine base pair is less stable than its deoxyguanosine counterpart in the same sequence environment. Implications of these results for design of DNA oligonucleotide probes are discussed.
- Published
- 1985
- Full Text
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21. Evidence for Z-form RNA by vacuum UV circular dichroism.
- Author
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Riazance JH, Baase WA, Johnson WC Jr, Hall K, Cruz P, and Tinoco I Jr
- Subjects
- Circular Dichroism, Ultraviolet Rays, Nucleic Acid Conformation, RNA
- Abstract
Circular dichroism (CD) spectra in the vacuum UV region for different conformations of poly d(G-C) X poly d(G-C) and poly r(G-C) X poly r(G-C) are very characteristic. The CD of the RNA in the A-form (6 M NaClO4 and 22 degrees C) is very similar to that of the DNA in 80% alcohol where it is believed to be in the A-form. With the exception of the longest wavelength transition, the CD of the RNA in 6 M NaClO4 at 46 degrees C is similar to the CD of the DNA under conditions where it is believed to be in the Z-form (2 M NaClO4). This substantiates that poly r(G-C) X poly r(G-C) assumes a left-handed Z-conformation in 6 M NaClO4 above 35 degrees C. CD spectra for the left-handed Z-forms of both the RNA and DNA are characterized by an intense negative peak at 190-195 nm, a crossover at about 184 nm, and an intense positive peak below 180 nm. The right-handed A- and B-forms of RNA and DNA all have an intense positive peak in their CD spectra near 186 nm. The large difference in CD in the range 185-195 nm for right- and left-handed conformations of nucleic acids can be used to identify the sense of helix winding.
- Published
- 1985
- Full Text
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22. Vacuum ultraviolet circular dichroism as an indicator of helical handedness in nucleic acids.
- Author
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Williams AL Jr, Cheong C, Tinoco I Jr, and Clark LB
- Subjects
- Adenine, Circular Dichroism methods, Cytosine, Guanine, Spectrophotometry, Ultraviolet methods, Thymine, Uracil, DNA, Nucleic Acid Conformation, RNA
- Abstract
Calculated circular dichroism spectra are presented for double-stranded polynucleotides of regular sequences in A-RNA, A-DNA, B-DNA, and Z-DNA conformations. Quantum mechanical matrix method calculations were carried out in the near and vacuum ultraviolet regions. In the near UV, the calculated spectra agreed qualitatively with the measured spectra. However in the far and vacuum UV, the calculated CD compared nearly quantitatively with the experimental spectra. The calculations show that the sign of the CD in the vacuum UV, in contrast to that in the near UV, can be correlated with the handedness of the helix.
- Published
- 1986
- Full Text
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23. The TFIIIA recognition fragment d(GGATGGGAG).d(CTCCCATCC) is B-form in solution.
- Author
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Aboul-ela F, Varani G, Walker GT, and Tinoco I Jr
- Subjects
- Binding Sites, Circular Dichroism, Magnetic Resonance Spectroscopy, Nucleic Acid Conformation, Transcription Factor TFIIIA, Deoxyribonucleotides metabolism, Transcription Factors metabolism
- Abstract
The deoxyoligonucleotide d(GGATGGGAG).d(CTCCCATCC) is a portion of the gene recognition sequence of transcription factor IIIA (TFIIIA). The crystal structure of this oligonucleotide was shown to be A-form (Mc Call, M., Brown, T., Hunter, W.N., and Kennard, O. 1986 Nature 322, 661-664). The present study employs NMR, optical, chemical and enzymatic techniques to investigate the solution structure of this DNA 9-mer. NMR COSY experiments indicate 16 of the 18 residues are predominantly south (C2'-endo) sugar conformation. NMR NOESY indicates glycosidic angles in the range predicted for B-form DNA as opposed to A-form. Related DNA and RNA self-complementary 18-mer sequences, d(GGATGGGAGC-TCCCATCC), with U substituted for T in RNA, were studied by circular dichroism. CD spectra support B-form structures for the DNA 9-mer and the DNA 18-mer, and A-form for the RNA 18-mer. High trifluoroethanol concentrations induce a B- to A-form transition in the DNA oligonucleotides. Enzymatic and chemical probes also illustrate significant differences between the DNA and the RNA oligonucleotides. We find no evidence to support an A-form conformation for the TFIIIA recognition sequence d(GGATGGGAG).d(CTCCCATCC) in solution.
- Published
- 1988
- Full Text
- View/download PDF
24. Secondary structure model for the complete simian virus 50 late precursor mRNA.
- Author
-
Nussinov R, Tinoco I Jr, and Jacobson AB
- Subjects
- Base Sequence, Computers, DNA Restriction Enzymes, Methods, Models, Biological, Nucleic Acid Conformation, DNA, Viral genetics, Genes, Viral, RNA, Messenger genetics, Simian virus 40 genetics
- Abstract
Structures for all sequences containing less than 1790 nucleotides in the 2600 nucleotide late region of the SV40 virus have been computed and saved on magnetic tape. Previously the longest sequence whose secondary structure was calculated in a single computer run contained 950 nucleotides. In the past, analysis of long molecules required numerous repeated, partially overlapping computations on much shorter segments. The structure obtained for the late half of the SV40 is Y-shaped with two unequal arms. It has 52 short hairpins. Two long range interactions between nucleotides near 650 and 1350 and between 1450 and 2450 appear to play an important role. The first is within the 16S intron; the second is in the 3' exon. The 5' and 3' ends of the molecule are close to each other and are found in the major elongated stem in the vicinity of the fork.
- Published
- 1982
- Full Text
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25. Thermodynamic studies of base pairing involving 2,6-diaminopurine.
- Author
-
Cheong C, Tinoco I Jr, and Chollet A
- Subjects
- 2-Aminopurine metabolism, Base Composition, Oligodeoxyribonucleotides metabolism, Thermodynamics, 2-Aminopurine analogs & derivatives, Adenine analogs & derivatives
- Abstract
The thermal stabilities of oligodeoxyribonucleotide duplexes containing 2,6-diaminopurine (D) matched with each of the four normal DNA bases were determined by optical melting techniques. Comparison of optical melting curves yielded relative stabilities for the D-containing standard base pairs in an otherwise identical base-pair sequence. The D:T pair was found to be more stable than the A:T pair in dC3DG3:dC3TG3, as stable as the A:T in dCT3DT3G:dCA3TA3G, and less stable than the A:T in dCA3DA3G:dCT7G. The order of stabilities for X:Y in the DNA duplex dCA3XA3G:dCT3YT3G is: (A:T) greater than (T:D) congruent to (D:T) greater than or equal to (T:A) greater than (C:D) congruent to (D:A) congruent to (D:G) greater than or equal to (D:C) congruent to (G:D) congruent to (D:D) greater than or equal to (A:D). Implications of these results for design of DNA oligonucleotide probes are discussed.
- Published
- 1988
- Full Text
- View/download PDF
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