Two initiation sites detected in the small s1 species of reovirus mRNA by dipeptide synthesis in vitro.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 81 ·No. 4 ·1984-02-00 ·Pages 1084-8

Cenatiempo Y, Twardowski T, Shoeman R, Ernst H, Brot N, Weissbach H, Shatkin AJ

Abstract

Reovirus mRNAs directed the synthesis of fMet dipeptides in a translation initiation system reconstituted from rabbit reticulocyte initiation and elongation factors, Artemia salina 80S ribosomes, yeast fMet-tRNAiMet and Escherichia coli3H-labeled aminoacyl tRNAs. As predicted from the GC(U,G) codon that follows the 5'-proximal AUG in half of the viral mRNA species, fMet-Ala was the predominant dipeptide product obtained in response to a mixture of mRNAs or to the separated size classes of medium (m) and small (s) mRNA. The four individual small mRNA species each directed the synthesis of an fMet dipeptide that was consistent with the utilization of the 5'-proximal AUG for initiation. In addition to fMet-Asp, the s1 mRNA also directed fMet-Glu synthesis indicative of initiation in a second reading frame at the 5'-penultimate AUG. The tripeptide fMet-Glu-Tyr was also synthesized from s1 mRNA, which further verified this second initiation site. mRNAs containing 5'-terminal GpppG were 10-15% as active as the corresponding m7G-capped templates. The dipeptide assay provides a rapid method for determining initiation sites in individual mRNAs or in mixtures of mRNAs.

MeSH Terms

Amino Acid Sequence Animals Artemia Base Sequence Dipeptides/genetics Kinetics Mammalian orthoreovirus 3/genetics Nucleic Acid Hybridization Oligopeptides/genetics Peptide Chain Initiation, Translational RNA, Messenger/genetics RNA, Viral/genetics Rabbits Reoviridae/genetics Reticulocytes/metabolism Ribosomes/metabolism

Chemicals

Dipeptides Oligopeptides RNA, Messenger RNA, Viral

Authors & Affiliations

7 authors, click to expand affiliations / ORCID

Cenatiempo Y

Twardowski T

Shoeman R

Ernst H

Brot N

Weissbach H

Shatkin A J

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33 references, click to expand

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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.81.4.1084

Published

1984-02-00

Pages

1084-8

Language

English

Region

United States

NLM ID

7505876

PMCID

PMC344769

Subset

External Links

PubMed source DOI: 10.1073/pnas.81.4.1084 PMC Full Text View journal details

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Two initiation sites detected in the small s1 species of reovirus mRNA by dipeptide synthesis in vitro.

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