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PMID: 8002564 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Mutations affecting two adjacent amino acid residues in the alpha subunit of RNA polymerase block transcriptional activation by the bacteriophage P2 Ogr protein.

Journal of bacteriology ·Vol. 176 ·No. 24 ·1994-12-00 ·Pages 7430-8

Ayers DJ, Sunshine MG, Six EW, Christie GE

Abstract

The bacteriophage P2 ogr gene product is a positive regulator of transcription from P2 late promoters. The ogr gene was originally defined by compensatory mutations that overcame the block to P2 growth imposed by a host mutation, rpoA109, in the gene encoding the alpha subunit of RNA polymerase. DNA sequence analysis has confirmed that this mutation affects the C-terminal region of the alpha subunit, changing a leucine residue at position 290 to a histidine (rpoAL290H). We have employed a reporter plasmid system to screen other, previously described, rpoA mutants for effects on activation of a P2 late promoter and have identified a second allele, rpoA155, that blocks P2 late transcription. This mutation lies just upstream of rpoAL290H, changing the leucine residue at position 289 to a phenylalanine (rpoAL289F). The effect of the rpoAL289F mutation is not suppressed by the rpoAL290H-compensatory P2 ogr mutation. P2 ogr mutants that overcome the block imposed by rpoAL289F were isolated and characterized. Our results are consistent with a direct interaction between Ogr and the alpha subunit of RNA polymerase and support a model in which transcription factor contact sites within the C terminus of alpha are discrete and tightly clustered.

Related Genes
rpoA
MeSH Terms
Alleles Base Sequence Chromosome Mapping Chromosomes, Bacterial DNA-Directed RNA Polymerases/genetics Enterobacteriaceae/enzymology,genetics,virology Escherichia coli/enzymology,genetics,virology Gene Expression Regulation, Bacterial Genes, Reporter Molecular Sequence Data Mutation Peptide Fragments/genetics Promoter Regions, Genetic/genetics Salmonella typhimurium/enzymology,genetics,virology Selection, Genetic Sequence Analysis Structure-Activity Relationship Transcription Factors/metabolism Transcription, Genetic Viral Proteins/metabolism
Chemicals
Peptide Fragments Transcription Factors Viral Proteins ogr protein, Enterobacteria phage P2 DNA-Directed RNA Polymerases
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Ayers D J
Department of Microbiology, Virginia Commonwealth University, Richmond.
Sunshine M G
Six E W
Christie G E
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Article Info
Journal
Journal of bacteriology
Abbr.
J Bacteriol
ISSN
0021-9193
DOI
10.1128/jb.176.24.7430-7438.1994
Published
1994-12-00
Pages
7430-8
Language
English
Region
United States
NLM ID
2985120R
PMCID
PMC197197
Subset
IM
Grants
NIGMS NIH HHS · R01 GM34651 · United States
External Links
PubMed source DOI: 10.1128/jb.176.24.7430-7438.1994 PMC Full Text View journal details
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