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Selection of circularly permuted ribozymes from Bacillus subtilis RNAse P by substrate binding
- Source :
- Biochemistry. 33:14207-14212
- Publication Year :
- 1994
- Publisher :
- American Chemical Society (ACS), 1994.
-
Abstract
- The effect of a single break in the phosphodiester backbone of Bacillus subtilis RNAse P RNA (P RNA) was examined using circular permutation analysis (CPA). This method reveals that many of the phosphodiester bonds in this catalytic RNA can be broken with little or no effect on substrate binding. Phosphate positions that show strong effects are located mostly in regions conserved among all RNAse P RNAs, or they are in regions known to interact directly with the pre-tRNA substrate. Two circularly permuted isomers of P RNA were constructed and analyzed in detail. The KM for both circularly permuted isomers is nearly identical to that of the wild-type P RNA. Since the KM of the P RNA is essentially the same as the binding constant to the substrate, this finding confirms the CPA results. The implications of backbone breakage are discussed with respect to folding and catalysis of the RNAse P RNA.
- Subjects :
- Hot Temperature
RNase P
Stereochemistry
Molecular Sequence Data
Bacillus subtilis
Polymerase Chain Reaction
Biochemistry
Ribonuclease P
Substrate Specificity
Microbiology
RNA, Transfer, Phe
Endoribonucleases
RNA Precursors
RNA, Catalytic
Cloning, Molecular
Base Sequence
biology
Chemistry
Ribozyme
Substrate (chemistry)
RNA
RNA, Circular
Circular permutation in proteins
biology.organism_classification
Binding constant
Kinetics
RNA, Bacterial
Phosphodiester bond
biology.protein
Nucleic Acid Conformation
Subjects
Details
- ISSN :
- 15204995 and 00062960
- Volume :
- 33
- Database :
- OpenAIRE
- Journal :
- Biochemistry
- Accession number :
- edsair.doi.dedup.....c6cade3cc9e37b8a11a20097f337e361