The C-terminal 12 amino acids of sigma(N) are required for structure and function.

Abstract:

:The sigma(N) protein is an alternative sigma subunit of bacterial RNA polymerase. We investigated the role of a 12-amino-acid "tail" at the C-terminus of Klebsiella pneumoniae sigma(N), which was predicted to be largely surface-exposed and to be mostly loop (that is not alpha-helical or beta-strand). Deletion of this tail from N-terminal hexahistidine-tagged sigma(N) led to loss of sigma(N)-dependent transcription activity in vivo. We overexpressed and purified this deletion-mutant protein for in vitro characterization. The purified deleted protein showed decreased RNA polymerase core- and DNA-binding activities compared to the full-length protein and transcription activity was greatly impaired. Furthermore, evidence from circular dichroism and protease digestion experiments together suggested that deletion of the C-terminus tail resulted in a loss of conformational constraint in the protein. We discuss a possible structural role for the 12 amino acids at the C-terminus of sigma(N).

journal_name

Arch Biochem Biophys

authors

Studholme DJ,Finn RD,Chaney MK,Buck M

doi

10.1006/abbi.1999.1426

subject

Has Abstract

pub_date

1999-11-15 00:00:00

pages

234-40

issue

2

eissn

0003-9861

issn

1096-0384

pii

S0003986199914261

journal_volume

371

pub_type

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