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PMID: 1986368 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Enzymatic aminoacylation of single-stranded RNA with an RNA cofactor.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 88 ·No. 1 ·1991-01-01 ·Pages 209-13

Musier-Forsyth K, Scaringe S, Usman N, Schimmel P

Abstract

A chemically synthesized single-stranded ribonucleotide tridecamer derived from the 3' end of Escherichia coli alanine tRNA can be charged with alanine in the presence of short complementary RNA oligonucleotides that form duplexes with the 3' fragment. Complementary 5' oligomers of 9, 8, 6, and 4 nucleotides all confer charging of the 3' fragment. Furthermore, in the presence of limiting 5' oligomer, greater than stoichiometric amounts of the single-stranded 3' acceptor fragment can be aminoacylated. This is due to a reiterative process of transient duplex formation followed by charging, dissociation of the 5' oligomer, and then rebinding to an uncharged single-stranded ribotridecamer so as to create another transient duplex substrate. Thus, a short RNA oligomer serves as a cofactor for a charging enzyme, and it thereby makes possible the aminoacylation of single-stranded RNA. These results expand possibilities for flexible routes to the development of early charging and coding systems.

MeSH Terms
Alanine-tRNA Ligase/metabolism Base Composition Base Sequence Escherichia coli/enzymology,genetics Kinetics Macromolecular Substances Molecular Sequence Data Nucleic Acid Conformation RNA, Transfer, Ala/genetics,metabolism Transcription, Genetic
Chemicals
Macromolecular Substances RNA, Transfer, Ala Alanine-tRNA Ligase
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Musier-Forsyth K
Department of Biology, Massachusetts Institute of Technology, Cambridge 02139.
Scaringe S
Usman N
Schimmel P
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.88.1.209
Published
1991-01-01
Pages
209-13
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC50779
Subset
IM
Grants
NIGMS NIH HHS · GM 15539 · United States
NIGMS NIH HHS · GM 37641 · United States
External Links
PubMed source DOI: 10.1073/pnas.88.1.209 PMC Full Text View journal details
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