Coupling nucleotide hydrolysis to transcription activation performance in a bacterial enhancer binding protein.

Abstract:

:The bacterial enhancer binding proteins (bEBP) are members of the AAA+ protein family and have a highly conserved 'DE' Walker B motif thought to be involved in the catalytic function of the protein with an active role in nucleotide hydrolysis. Based on detailed structural data, we analysed the functionality of the conserved 'DE' Walker B motif of a bEBP model, phage shock protein F (PspF), to investigate the role of these residues in the sigma(54)-dependent transcription activation process. We established their role in the regulation of PspF self-association and in the relay of the ATPase activity to the remodelling of an RNA polymerase.promoter complex (Esigma(54).DNA). Specific substitutions of the conserved glutamate (E) allowed the identification of new functional ATP.bEBP.Esigma(54) complexes which are stable and transcriptionally competent, providing a new tool to study the initial events of the sigma(54)-dependent transcription activation process. In addition, we show the importance of this glutamate residue in sigma(54).DNA conformation sensing, permitting the identification of new intermediate stages within the transcription activation pathway.

journal_name

Mol Microbiol

journal_title

Molecular microbiology

authors

Joly N,Rappas M,Wigneshweraraj SR,Zhang X,Buck M

doi

10.1111/j.1365-2958.2007.05901.x

subject

Has Abstract

pub_date

2007-11-01 00:00:00

pages

583-95

issue

3

eissn

0950-382X

issn

1365-2958

pii

MMI5901

journal_volume

66

pub_type

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