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PMID: 6203741 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Simian virus 40 cRNA is processed into functional mRNA in microinjected monkey cells.

The EMBO journal ·Vol. 1 ·No. 9 ·1982-00-00 ·Pages 1081-8

Graessmann M, Graessmann A

Abstract

Monkey cells, microinjected with simian virus 40 (SV40) in vitro synthesized cRNA produce full-size tumor (T)-antigen. This was verified by analyzing immunoprecipitates of microinjected cells by polyacrylamide gel electrophoresis. Early SV40 DNA contains an intron within the large T-antigen coding sequences. Therefore, cRNA copied in vitro from the early DNA strand requires removal of the intron in order to become a functional mRNA. Polyadenylation of the cRNA in vitro by Escherichia coli poly(A)-polymerase increased the biological activity of the RNA. Detection of T-antigen by gel electrophoresis required as little as 50 poly(A)-cRNA injected cells. Splicing of the microinjected cRNA appears to be a nuclear process. Cells enucleated by cytochalasin B prior to injection do not synthesize large T-antigen. However, small t-antigen, a protein with a continuous sequence, is synthesized in these cells. Finally, it is shown that the process of splicing is not required for the transport of mRNA from the nucleus into the cytoplasm. Authentic T-antigen mRNA, isolated from virus infected cells, induced T-antigen synthesis with similar efficiency after either nuclear or cytoplasmic injection.

MeSH Terms
Animals Antigens, Polyomavirus Transforming Base Sequence Cell Line Cell Transformation, Viral Chlorocebus aethiops DNA Restriction Enzymes Genes, Viral Lung Microinjections Protein Kinases/genetics RNA/genetics RNA, Complementary RNA, Messenger/genetics Rats Simian virus 40/genetics Viral Proteins/genetics
Chemicals
Antigens, Polyomavirus Transforming RNA, Complementary RNA, Messenger Viral Proteins RNA Protein Kinases DNA Restriction Enzymes
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Graessmann M
Graessmann A
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32 references, click to expand
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Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
Published
1982-00-00
Pages
1081-8
Language
English
Region
England
NLM ID
8208664
PMCID
PMC553166
Subset
IM
External Links
PubMed source PMC Full Text View journal details
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