CAP interacts with RNA polymerase in solution in the absence of promoter DNA

Tomasz Heyduk, James Lee, Yon W. Ebright, Erich E. Blatter, Yuhong Zhou, Richard H. Ebright

Research output: Contribution to journalArticle

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Abstract

PROTEIN-PROTEIN interactions between transcription activator proteins and RNA polymerase or basal transcription factors have been suggested to be important for transcription activation1-8. Interactions between catabolite gene activator protein (CAP)9,10 and RNA polymerase have been proposed based on face-of-helix-dependent transcription activation by CAP11-13 and based on face-of-helix-dependent cooperative binding of CAP and RNA polymerase to promoter DNA14,15. Mutants of CAP specifically defective in transcription activation have been isolated (mutants defective in transcription activation, but not defective in DNA binding and DNA bending16-19). All such mutants contain amino-acid substitutions within a surface loop consisting of amino acids 152 to 166 of CAP16-19. Here we use the thermodynamically rigorous technique of fluorescence polarization20-23 to show that CAP interacts with RNA polymerase in solution in the absence of promoter DNA (KD,app = 2.8 × 10-7 M), whereas [Ala158]CAP, a mutant of CAP specifically defective in transcription activation, does not.

Original languageEnglish (US)
Pages (from-to)548-549
Number of pages2
JournalNature
Volume364
Issue number6437
StatePublished - Aug 5 1993

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Cyclic AMP Receptor Protein
DNA-Directed RNA Polymerases
Transcriptional Activation
DNA
Amino Acid Substitution
Transcription Factors
Fluorescence
Amino Acids

ASJC Scopus subject areas

  • General

Cite this

Heyduk, T., Lee, J., Ebright, Y. W., Blatter, E. E., Zhou, Y., & Ebright, R. H. (1993). CAP interacts with RNA polymerase in solution in the absence of promoter DNA. Nature, 364(6437), 548-549.

CAP interacts with RNA polymerase in solution in the absence of promoter DNA. / Heyduk, Tomasz; Lee, James; Ebright, Yon W.; Blatter, Erich E.; Zhou, Yuhong; Ebright, Richard H.

In: Nature, Vol. 364, No. 6437, 05.08.1993, p. 548-549.

Research output: Contribution to journalArticle

Heyduk, T, Lee, J, Ebright, YW, Blatter, EE, Zhou, Y & Ebright, RH 1993, 'CAP interacts with RNA polymerase in solution in the absence of promoter DNA', Nature, vol. 364, no. 6437, pp. 548-549.
Heyduk T, Lee J, Ebright YW, Blatter EE, Zhou Y, Ebright RH. CAP interacts with RNA polymerase in solution in the absence of promoter DNA. Nature. 1993 Aug 5;364(6437):548-549.
Heyduk, Tomasz ; Lee, James ; Ebright, Yon W. ; Blatter, Erich E. ; Zhou, Yuhong ; Ebright, Richard H. / CAP interacts with RNA polymerase in solution in the absence of promoter DNA. In: Nature. 1993 ; Vol. 364, No. 6437. pp. 548-549.
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