Structural and Biophysical Studies on Two Promoter Recognition Domains of the Extra-cytoplasmic Function σ Factor σC from Mycobacterium tuberculosis

Krishan Gopal Thakur, Anagha Madhusudan Joshi, B. Gopal

Research output: Contribution to journalArticlepeer-review

Abstract

σ factors are transcriptional regulatory proteins that bind to the RNA polymerase and dictate gene expression. The extracytoplasmic function (ECF) σ factors govern the environment dependent regulation of transcription. ECF σ factors have two domains σ2 and σ4 that recognize the -10 and -35 promoter elements. However, unlike the primary σ factor σA, the ECF σ factors lack σ3, a region that helps in the recognition of the extended -10 element and σ1.1, a domain involved in the autoinhibition of σA in the absence of core RNA polymerase. Mycobacterium tuberculosis σC is an ECF σ factor that is essential for the pathogenesis and virulence of M. tuberculosis in the mouse and guinea pig models of infection. However, unlike other ECF σ factors, σC does not appear to have a regulatory anti-σ factor located in the same operon. We also note that M. tuberculosis σC differs from the canonical ECF σ factors as it has an N-terminal domain comprising of 126 amino acids that precedes the σC2 and σC4 domains. In an effort to understand the regulatory mechanism of this protein, the crystal structures of the σC2 and σC4 domains of σC were determined. These promoter recognition domains are structurally similar to the corresponding domains of σA despite the low sequence similarity. Fluorescence experiments using the intrinsic tryptophan residues of σC2 as well as surface plasmon resonance measurements reveal that the σC2 and σC4 domains interact with each other. Mutational analysis suggests that the Pribnow box-binding region of σC2 is involved in this interdomain interaction. Interaction between the promoter recognition domains in M. tuberculosis σC are thus likely to regulate the activity of this protein even in the absence of an anti-σ factor.
Original languageEnglish
Pages (from-to)4711-4718
Number of pages8
JournalJournal of Biological Chemistry
Volume282
Issue number7
DOIs
Publication statusPublished - 2007

Keywords

  • ANIMALS
  • Bacterial Proteins
  • Crystallography, X-Ray
  • Cytoplasm
  • DNA, Bacterial
  • DNA-Directed RNA Polymerases
  • Disease Models, Animal
  • Gene Expression Regulation, Bacterial
  • Guinea Pigs
  • HUMANS
  • MICE
  • Models, Molecular
  • MUTATION
  • Mycobacterium tubercolosis
  • Promoter Regions, Genetic
  • Protein Binding
  • protein structure
  • Sigma Factor
  • Surface Plasmon Resonance
  • Transcription, Genetic
  • Tubercolosis

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