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PMID: 11345430 Published · ppublish English Journal Article

Optimization and optimality of a short ribozyme ligase that joins non-Watson-Crick base pairings.

RNA (New York, N.Y.) ·Vol. 7 ·No. 4 ·2001-04-00 ·Pages 513-23

Robertson MP, Hesselberth JR, Ellington AD

Abstract

A small ribozyme ligase (L1) selected from a random sequence population appears to utilize non-Watson-Crick base pairs at its ligation junction. Mutational and selection analyses confirmed the presence of these base pairings. Randomization of the L1 core and selection of active ligases yielded highly active variants whose rates were on the order of 1 min(-1). Base-pairing covariations confirmed the general secondary structure of the ligase, and the most active ligases contained a novel pentuple sequence covariation. The optimized L1 ligases may be optimal within their sequence spaces, and minimal ligases that span less than 60 nt in length have been engineered based on these results.

MeSH Terms
Base Pairing Base Sequence Catalysis Directed Molecular Evolution Ligases Models, Molecular Molecular Sequence Data Nucleic Acid Conformation RNA, Catalytic/genetics,metabolism Substrate Specificity
Chemicals
RNA, Catalytic Ligases
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Robertson M P
Department of Chemistry and Biochemistry, Institute for Cellular and Molecular Biology, University of Texas at Austin, 78712, USA.
Hesselberth J R
Ellington A D
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Article Info
Journal
RNA (New York, N.Y.)
Abbr.
RNA
ISSN
1355-8382
DOI
10.1017/s1355838201002199
Published
2001-04-00
Pages
513-23
Language
English
Region
United States
NLM ID
9509184
PMCID
PMC1370105
Subset
IM
External Links
PubMed source DOI: 10.1017/s1355838201002199 PMC Full Text View journal details
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