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PMID: 8392187 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Identification of the activating region of catabolite gene activator protein (CAP): isolation and characterization of mutants of CAP specifically defective in transcription activation.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 90 ·No. 13 ·1993-07-01 ·Pages 6081-5

Zhou Y, Zhang X, Ebright RH

Abstract

We have isolated 21 mutants of catabolite gene activator protein (CAP) defective in transcription activation at the lac promoter but not defective in DNA binding. The amino acid substitutions in the mutants map to a single region of CAP: amino acids 156-162. As assessed in vitro, the substituted CAP variants are nearly completely unable to activate transcription at the lac promoter but bind to DNA with the same affinity and bend DNA to the same extent as wild-type CAP. Our results establish that amino acids 156-162 are critical for transcription activation at the lac promoter but not for DNA binding and DNA bending. In the structure of CAP, amino acids 156-162 are part of a surface loop. We propose that this surface loop makes a direct protein-protein contact with RNA polymerase at the lac promoter.

Related Genes
crp
MeSH Terms
Base Sequence Cyclic AMP Receptor Protein/chemistry,genetics,physiology DNA/chemistry,metabolism DNA-Directed RNA Polymerases/metabolism Lac Operon Models, Molecular Molecular Sequence Data Mutation Nucleic Acid Conformation Peptide Fragments/physiology Promoter Regions, Genetic Structure-Activity Relationship Transcriptional Activation
Chemicals
Cyclic AMP Receptor Protein Peptide Fragments DNA DNA-Directed RNA Polymerases
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Zhou Y
Department of Chemistry, Rutgers University, New Brunswick, NJ 08855.
Zhang X
Ebright R H
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26 references, click to expand
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.90.13.6081
Published
1993-07-01
Pages
6081-5
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC46871
Subset
IM
Grants
NIGMS NIH HHS · GM41376 · United States
External Links
PubMed source DOI: 10.1073/pnas.90.13.6081 PMC Full Text View journal details
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