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PMID: 7984416 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Start site selection at lacUV5 promoter affected by the sequence context around the initiation sites.

Nucleic acids research ·Vol. 22 ·No. 22 ·1994-11-11 ·Pages 4667-72

Jeong W, Kang C

Abstract

The effects of single base pair substitutions at the initiation sites of lacUV5 promoter on the transcription start site selection by E. coli RNA polymerase were systematically studied. Transcription start sites were mapped by sizing the cytosine-specifically terminated transcripts produced in vitro by using a chain terminator 3'-deoxycytidine 5'-triphosphate (3'-dCTP) in transcription reactions. Transcription of a prototype lacUV5 promoter initiated with three purines (-1G, +1A and +2A; +1 representing the predominant start site) located 6-8 bp downstream from the Pribnow box. All the substitutions affected the start site selection, resulting in a change in the number of start sites (from 3 to 2 or 1) and/or a shift of the major start site (to -1 or +2). None of the variants started outside the 3-bp region and at the positions substituted by a pyrimidine. Purine-to-pyrimidine changes suppressed not only initiation at the substituted position but also, in some cases, at the other purine position. Purine-to-purine changes also shifted the major start site or suppressed the initiation at other sites. Changes at -2 and +5 also affected the start site selection. Thus, the sequence context around the initiation sites of lacUV5 promoter strongly influences the selection of initiating nucleotides by E. coli RNA polymerase.

Related Genes
lacUV5
MeSH Terms
Base Sequence DNA-Directed RNA Polymerases Escherichia coli/enzymology,genetics Lac Operon/genetics Molecular Sequence Data Point Mutation/genetics Promoter Regions, Genetic/genetics RNA, Messenger/biosynthesis Transcription, Genetic/genetics
Chemicals
RNA, Messenger DNA-Directed RNA Polymerases
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Jeong W
Department of Life Science, Korea Advanced Institute of Science and Technology, Yusong-gu, Taejon.
Kang C
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28 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/22.22.4667
Published
1994-11-11
Pages
4667-72
Language
English
Region
England
NLM ID
0411011
PMCID
PMC308516
Subset
IM
External Links
PubMed source DOI: 10.1093/nar/22.22.4667 PMC Full Text View journal details
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