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PMID: 8892810 Published · ppublish English Journal Article

An N-terminal mutation in the bacteriophage T4 motA gene yields a protein that binds DNA but is defective for activation of transcription.

Journal of bacteriology ·Vol. 178 ·No. 21 ·1996-11-00 ·Pages 6133-9

Gerber JS, Hinton DM

Abstract

The bacteriophage T4 MotA protein is a transcriptional activator of T4-modified host RNA polymerase and is required for activation of the middle class of T4 promoters. MotA alone binds to the -30 region of T4 middle promoters, a region that contains the MotA box consensus sequence [(t/a)(t/a)TGCTT(t/c)A]. We report the isolation and characterization of a protein designated Mot21, in which the first 8 codons of the wild-type motA sequence have been replaced with 11 different codons. In gel retardation assays, Mot21 and MotA bind DNA containing the T4 middle promoter P(uvsX) similarly, and the proteins yield similar footprints on P(uvsX). However, Mot21 is severely defective in the activation of transcription. On native protein gels, a new protein species is seen after incubation of the sigma70 subunit of RNA polymerase and wild-type MotA protein, suggesting a direct protein-protein contact between MotA and sigma70. Mot21 fails to form this complex, suggesting that this interaction is necessary for transcriptional activation and that the Mot21 defect arises because Mot21 cannot form this contact like the wild-type activator.

MeSH Terms
Bacteriophage T4/genetics,metabolism Base Sequence DNA, Viral DNA-Binding Proteins/genetics,isolation & purification,metabolism DNA-Directed RNA Polymerases/metabolism Escherichia coli/metabolism Genetic Complementation Test Membrane Proteins/genetics Molecular Sequence Data Promoter Regions, Genetic Sigma Factor/metabolism Transcription Factors/genetics,isolation & purification,metabolism Transcriptional Activation Viral Proteins/genetics,isolation & purification,metabolism
Chemicals
DNA, Viral DNA-Binding Proteins Membrane Proteins MotA protein, Enterobacteria phage T4 Sigma Factor Transcription Factors UvsX protein, Enterobacteria phage T4 Viral Proteins RNA polymerase sigma 70 DNA-Directed RNA Polymerases
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Gerber J S
Section on Nucleic Acid Biochemistry, National Institute of Diabetes and Digestive and Kidney Diseases, Bethesda, Maryland 20892, USA.
Hinton D M
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Article Info
Journal
Journal of bacteriology
Abbr.
J Bacteriol
ISSN
0021-9193
DOI
10.1128/jb.178.21.6133-6139.1996
Published
1996-11-00
Pages
6133-9
Language
English
Region
United States
NLM ID
2985120R
PMCID
PMC178481
Subset
IM
External Links
PubMed source DOI: 10.1128/jb.178.21.6133-6139.1996 PMC Full Text View journal details
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