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PMID: 3005968 Published · ppublish English Journal Article Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S.

In vitro splicing of simian virus 40 early pre mRNA.

Nucleic acids research ·Vol. 14 ·No. 3 ·1986-02-11 ·Pages 1219-35

Noble JC, Prives C, Manley JL

Abstract

The products of splicing of simian virus 40 early pre mRNA in HeLa cell nuclear extracts have been characterized. Of the two alternative splicing patterns exhibited by this precursor in vivo, which involve the use of alternative large T and small t 5' splice sites and a single shared 3' splice site, only one, producing large T mRNA, was found to occur in vitro. A number of possible intermediates and byproducts of splicing of large T mRNA were observed, including free large T 5' exon, lariat form intron joined to 3' exon and free lariat and linear forms of large T intron. The formation of these products argues strongly for a basic similarity in the mechanism underlying large T and other, non-alternative splices. A collection of RNAs resulting from protection of early pre mRNA at specific points from an endogenous 5' to 3' exonuclease activity in vitro have also been observed. The regions of the precursor RNA protected map to positions immediately upstream of the 5' splice sites of large T and small t and the lariat branchpoint, and may represent interaction of these regions with components of the splicing machinery.

MeSH Terms
Antigens, Viral, Tumor/genetics Base Sequence Cell Nucleus/metabolism Cell-Free System DNA, Viral/genetics Electrophoresis, Polyacrylamide Gel Endonucleases HeLa Cells Kinetics Nucleic Acid Precursors/genetics RNA Splicing RNA, Messenger/genetics RNA, Viral/genetics Simian virus 40/genetics Single-Strand Specific DNA and RNA Endonucleases
Chemicals
Antigens, Viral, Tumor DNA, Viral Nucleic Acid Precursors RNA, Messenger RNA, Viral Endonucleases Single-Strand Specific DNA and RNA Endonucleases
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Noble J C
Prives C
Manley J L
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48 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/14.3.1219
Published
1986-02-11
Pages
1219-35
Language
English
Region
England
NLM ID
0411011
PMCID
PMC339499
Subset
IM
Grants
NCI NIH HHS · CA 33620 · United States
Databases
GENBANK
J02400, J02402, J02403, J02406, J02407, J02408, J02409, J02410, V01380
External Links
PubMed source DOI: 10.1093/nar/14.3.1219 PMC Full Text View journal details
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