Cooperative DNA binding of heterologous proteins: Evidence for contact between the cyclic AMP receptor protein and RNA polymerase

Journal Article · Wed Jun 01 00:00:00 EDT 1988 · Proceedings of the National Academy of Sciences of the United States of America; (USA)

DOI:https://doi.org/10.1073/pnas.85.12.4138· OSTI ID:5642350

Ren, Y L; Garges, S; Adhya, S; Krakow, J S ^[1]

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1. Hunter College of the City Univ. of New York, NY (USA)

Four cAMP-independent receptor protein mutants (designated CRP* mutants) isolated previously are able to activate in vivo gene transcription in the absence of cAMP and their activity can be enhanced by cAMP or cGMP. One of the four mutant proteins, CRP*598 (Arg-142 to His, Ala-144 to Thr), has been characterized with regard to its conformational properties and ability to bind to and support abortive initiation from the lac promoter. Binding of wild-type CRP to its site on the lac promoter and activation of abortive initiation by RNA polymerase on this promoter are effected by cAMP but not by cGMP. CRP*598 can activate lacP{sup +}-directed abortive initiation in the presence of cAMP and less efficiently in the presence of cGMP or in the absence of cyclic nucleotide. DNase I protection (footprinting) indicates that cAMP-CRP* binds to its site on the lac promoter whereas unliganded CRP* and cGMP-CRP* form a stable complex with the ({sup 32}P)lacP{sup +} fragment only in the presence of RNA polymerase, showing cooperative binding of two heterologous proteins. This cooperative binding provides strong evidence for a contact between CRP and RNA polymerase for activation of transcription. Although cGMP binds to CRP, it cannot replace cAMP in effecting the requisite conformational transition necessary for site-specific promoter binding.

• OSTI ID: 5642350
• Journal Information: Proceedings of the National Academy of Sciences of the United States of America; (USA), Vol. 85:12; ISSN 0027-8424
• Country of Publication: United States
• Language: English

Related Subjects

59 BASIC BIOLOGICAL SCIENCES
PROTEINS
CROSS-LINKING
RECEPTORS
CONFORMATIONAL CHANGES
RNA POLYMERASES
TRANSCRIPTION
GENE REGULATION
AMP
DNA-ASE
ESCHERICHIA COLI
MUTANTS
PHOSPHORUS 32
TRITIUM COMPOUNDS
BACTERIA
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
CHEMICAL REACTIONS
DAYS LIVING RADIOISOTOPES
ENZYMES
ESTERASES
HYDROGEN COMPOUNDS
HYDROLASES
ISOTOPES
LIGHT NUCLEI
MEMBRANE PROTEINS
MICROORGANISMS
NUCLEI
NUCLEOTIDES
NUCLEOTIDYLTRANSFERASES
ODD-ODD NUCLEI
ORGANIC COMPOUNDS
PHOSPHODIESTERASES
PHOSPHORUS ISOTOPES
PHOSPHORUS-GROUP TRANSFERASES
POLYMERASES
POLYMERIZATION
RADIOISOTOPES
TRANSFERASES
550201* - Biochemistry- Tracer Techniques