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PMID: 7029468 Published · ppublish English Journal Article

Separation of the plus and minus strands of cytoplasmic polyhedrosis virus and human reovirus double-stranded genome RNAs by gel electrophoresis.

Nucleic acids research ·Vol. 9 ·No. 20 ·1981-10-24 ·Pages 5269-86

Smith RE, Morgan MA, Furuichi Y

Abstract

The complementary strands of most of the genome double-stranded RNA segments of insect cytoplasmic polyhedrosis virus (CPV) and human reovirus are separated for the first time by agarose gel electrophoresis in in the presence of 7 M urea. CPV (+) strands and most reovirus (-) strands migrate faster than the corresponding strands of opposite polarity. Glyoxal treatment, which modifies guanine residues and prevents G-C basepairing, results in a loss of strand resolution and concomitantly a significant decrease in electrophoretic mobilities. Reovirus mRNAs synthesized in vitro with ITP substituted for GTP show similar decreased electrophoretic mobilities as the glyoxalated mRNAs. These results clearly indicate that the basis for (+) and (-) strand resolution is the presence of secondary structure formed mainly by G-C(U) base-pairs that are maintained during gel electrophoresis in the presence of 7 M urea. When the plus and minus strands of CPV genomes were separated and compared for protein synthesizing activity, it was found that only the plus strands were able to form stable 80S ribosome-RNA initiation complexes in wheat germ cell-free extracts.

MeSH Terms
Base Composition Base Sequence Genes, Viral Humans Insect Viruses/genetics Plants/metabolism RNA, Double-Stranded/isolation & purification RNA, Messenger/genetics RNA, Viral/isolation & purification Reoviridae/genetics Ribosomes/metabolism Triticum/metabolism
Chemicals
RNA, Double-Stranded RNA, Messenger RNA, Viral
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Smith R E
Morgan M A
Furuichi Y
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33 references, click to expand
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Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
0305-1048
DOI
10.1093/nar/9.20.5269
Published
1981-10-24
Pages
5269-86
Language
English
Region
England
NLM ID
0411011
PMCID
PMC327519
Subset
IM
External Links
PubMed source DOI: 10.1093/nar/9.20.5269 PMC Full Text View journal details
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