NDLI: A sigma32 mutant with a single amino acid change in the highly conserved region 2.2 exhibits reduced core RNA polymerase affinity.

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A sigma32 mutant with a single amino acid change in the highly conserved region 2.2 exhibits reduced core RNA polymerase affinity.

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A sigma32 mutant with a single amino acid change in the highly conserved region 2.2 exhibits reduced core RNA polymerase affinity.

Content Provider	World Health Organization (WHO)-Global Index Medicus
Author	Ng, N. Calendar, R. Joo, D. M.
Description	Author Affiliation: Joo DM ( Department of Molecular and Cell Biology, University of California, Berkeley, CA 94720, USA.);
Abstract	sigma32, the product of the rpoH gene in Escherichia coli, provides promoter specificity by interacting with core RNAP. Amino acid sequence alignment of sigma32 with other sigma factors in the sigma70 family has revealed regions of sequence homology. We have investigated the function of the most highly conserved region, 2.2, using purified products of various rpoH alleles. Core RNAP binding analysis by glycerol gradient sedimentation has revealed reduced core RNAP affinity for one of the mutant sigma32 proteins, Q80R. This reduced core interaction is exacerbated in the presence of sigma70, which competes with sigma32 for binding of core RNAP. When a different but more conserved amino acid was introduced at this position by site-directed mutagenesis (Q80N), this mutant sigma factor still displayed a significant reduction in its core RNAP affinity. Based on these results, we conclude that at least one specific amino acid in region 2.2 is involved in core RNAP interaction.
ISSN	00278424
e-ISSN	10916490
Journal	Proceedings of the National Academy of Sciences of the United States of America
Issue Number	10
Volume Number	94
Language	English
Publisher	National Academy of Sciences
Publisher Date	1997-06-01
Publisher Place	United States
Access Restriction	Open
Subject Keyword	DNA-Directed RNA Polymerases Metabolism Escherichia Coli Proteins Escherichia Coli Heat-Shock Proteins Protein Conformation Sigma Factor Transcription Factors Amino Acid Sequence Bacterial Proteins Chemistry Isolation & Purification Binding Sites Cloning, Molecular Conserved Sequence Genetics HSP70 Heat-Shock Proteins Biosynthesis Kinetics Models, Molecular Mutagenesis, Site-Directed Point Mutation Recombinant Proteins Transcription, Genetic Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S. Multidisciplinary
Content Type	Text
Resource Type	Article
Subject	Multidisciplinary

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