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PMID: 7835347 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Efficient trans-cleavage of a stem-loop RNA substrate by a ribozyme derived from neurospora VS RNA.

The EMBO journal ·Vol. 14 ·No. 2 ·1995-01-16 ·Pages 368-76

Guo HC, Collins RA

Abstract

We have constructed a ribozyme containing 144 nucleotides of Neurospora VS RNA that can catalyze the cleavage of a separate RNA in a true enzymatic manner (Km approximately 0.13 microM, kcat approximately 0.7/min). Comparison of the rates of cis- and trans-cleavage, as well as the lack of effect of pH on the rate of cleavage, suggest that a rate-limiting step, possibly a conformational change, occurs prior to cleavage. The minimum contiguous substrate sequence required for cleavage consists of one nucleotide upstream and 19 nucleotides downstream of the cleavage site. Unlike most other ribozymes which interact with long single-stranded regions of their substrates, the minimal substrate for the VS ribozyme consists mostly of a stable stem-loop, which would appear to preclude its recognition simply via extensive Watson-Crick base pairing.

MeSH Terms
Base Sequence Hydrogen-Ion Concentration Hydrolysis Molecular Sequence Data Neurospora/genetics,metabolism RNA, Catalytic/metabolism RNA, Fungal/chemistry,metabolism Substrate Specificity
Chemicals
RNA, Catalytic RNA, Fungal
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Guo H C
Canadian Institute for Advanced Research Program in Evolutionary Biology, University of Toronto, Ontario.
Collins R A
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Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
Published
1995-01-16
Pages
368-76
Language
English
Region
England
NLM ID
8208664
PMCID
PMC398091
Subset
IM
External Links
PubMed source PMC Full Text View journal details
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