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PMID: 140861 Published · ppublish English Journal Article Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S.

Characterization and in vitro translation of polyadenylated messenger ribonucleic acid from Neurospora crassa.

Journal of bacteriology ·Vol. 130 ·No. 3 ·1977-06-00 ·Pages 1192-8

Lucas MC, Jacobson JW, Giles NH

Abstract

Ribonucleic acid (RNA) extracted from Neurospora crassa has been fractionated by oligodeoxythymidylic acid [oligo(dT)]-cellulose chromatography into polyadenylated messenger RNA [poly(A) mRNA] and unbound RNA. The poly(A) mRNA, which comprises approximately 1.7% of the total cellular RNA, was further characterized by Sepharose 4B chromatography and polyacrylamide gel electrophoresis. Both techniques showed that the poly(A) mRNA was heterodisperse in size, with an average molecular weight similar to that of 17S ribosomal RNA (rRNA). The poly(A) segments isolated from the poly(A) mRNA were relatively short, with three major size classes of 30, 55, and 70 nucleotides. Gel electrophoresis of the non-poly(A) RNA indicated that it contained primarily rRNA and 4S RNA. The optimal conditions were determined for the translation of Neurospora mRNA in a cell-free wheat germ protein-synthesizing system. Poly(A) mRNA stimulated the incorporation of [14C]leucine into polypeptides ranging in size from 10,000 to 100,000 daltons. The RNA that did not bind to oligo(dT)-cellulose also stimulated the incorporation of [14C]leucine, indicating that this fraction contains a significant concentration of mRNA which has either no poly(A) or very short poly(A) segments. In addition, the translation of both poly(A) mRNA and unbound mRNA was inhibited by 7-methylguanosine-5'-monophosphate (m7G5'p). This is preliminary evidence for the existence of a 5'-RNA "cap" on Neurospora mRNA.

MeSH Terms
Chromatography, Agarose Chromatography, Ion Exchange Electrophoresis, Polyacrylamide Gel Neurospora/metabolism Neurospora crassa/analysis,metabolism Poly A/isolation & purification,metabolism Protein Biosynthesis RNA, Messenger/isolation & purification,metabolism
Chemicals
RNA, Messenger Poly A
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Lucas M C
Jacobson J W
Giles N H
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33 references, click to expand
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Article Info
Journal
Journal of bacteriology
Abbr.
J Bacteriol
ISSN
0021-9193
DOI
10.1128/jb.130.3.1192-1198.1977
Published
1977-06-00
Pages
1192-8
Language
English
Region
United States
NLM ID
2985120R
PMCID
PMC235342
Subset
IM
External Links
PubMed source DOI: 10.1128/jb.130.3.1192-1198.1977 PMC Full Text View journal details
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