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PMID: 6174941 Published · ppublish English Journal Article

Comparison of the polyoma virus early and late promoters by transcription in vitro.

Nucleic acids research ·Vol. 10 ·No. 3 ·1982-02-11 ·Pages 871-87

Jat P, Roberts JW, Cowie A, Kamen R

Abstract

Polyoma virus DNA was transcribed in the HeLa whole cell extract in vitro system (1). Early region transcripts with the same 5'-ends as in vivo mRNAs, located 31 +/- 2bp from 'TATA'-boxes, were synthesized by RNA polymerase II. Sequences sufficient for efficient expression of the early promoter were present in a substitution mutant lacking viral DNA from a position 55bp before the principal cap sites. Late region transcripts were synthesised inefficiently. Only one (at nt5129 +/- 2) of the many late mRNA cap sites functioned as an in vitro initiation point. This was the one 5'-end located 31 +/- 2bp from a sequence resembling the 'TATA' consensus. The proportion of late to early region RNA polymerase II transcripts decreased dramatically at suboptimal template concentrations. An hypothesis to explain the regulation of late gene expression in vivo based on these results is proposed. A linear templates were transcribed only by RNA polymerase II, transcripts with the same sense as late mRNAs and 5'-ends at nt5076 +/- 2 were produced from superhelical template by an alpha amanitin resistant enzyme.

MeSH Terms
Base Sequence DNA Restriction Enzymes DNA, Viral/genetics HeLa Cells/metabolism Humans Kinetics Operon Polyomavirus/genetics RNA Polymerase II/metabolism RNA, Messenger/genetics RNA-Directed DNA Polymerase/metabolism Transcription, Genetic
Chemicals
DNA, Viral RNA, Messenger RNA Polymerase II RNA-Directed DNA Polymerase DNA Restriction Enzymes
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Jat P
Roberts J W
Cowie A
Kamen R
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44 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/10.3.871
Published
1982-02-11
Pages
871-87
Language
English
Region
England
NLM ID
0411011
PMCID
PMC326208
Subset
IM
Databases
GENBANK
J02288, J02290, J02291, J02292, K00932, K00997, K01041, K01071, K01072, V01117, V01147
External Links
PubMed source DOI: 10.1093/nar/10.3.871 PMC Full Text View journal details
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