Structure and function of RNase AS, a polyadenylate-specific exoribonuclease affecting mycobacterial virulence in vivo

Structure. 2014 May 6;22(5):719-30. doi: 10.1016/j.str.2014.01.014. Epub 2014 Apr 3.

Abstract

The cell-envelope of Mycobacterium tuberculosis plays a key role in bacterial virulence and antibiotic resistance. Little is known about the molecular mechanisms of regulation of cell-envelope formation. Here, we elucidate functional and structural properties of RNase AS, which modulates M. tuberculosis cell-envelope properties and strongly impacts bacterial virulence in vivo. The structure of RNase AS reveals a resemblance to RNase T from Escherichia coli, an RNase of the DEDD family involved in RNA maturation. We show that RNase AS acts as a 3'-5'-exoribonuclease that specifically hydrolyzes adenylate-containing RNA sequences. Also, crystal structures of complexes with AMP and UMP reveal the structural basis for the observed enzyme specificity. Notably, RNase AS shows a mechanism of substrate recruitment, based on the recognition of the hydrogen bond donor NH2 group of adenine. Our work opens a field for the design of drugs able to reduce bacterial virulence in vivo.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenine
  • Adenosine Monophosphate / chemistry
  • Adenosine Monophosphate / metabolism
  • Animals
  • Bacterial Proteins / chemistry
  • Bacterial Proteins / metabolism
  • Crystallography, X-Ray
  • Embryo, Nonmammalian / microbiology
  • Exoribonucleases / chemistry
  • Gene Knockout Techniques
  • Hydrogen Bonding
  • Models, Molecular
  • Mutation
  • Mycobacterium marinum / genetics
  • Mycobacterium marinum / pathogenicity
  • Mycobacterium tuberculosis / enzymology
  • Mycobacterium tuberculosis / pathogenicity*
  • Poly A / metabolism
  • Protein Multimerization
  • Ribonucleases / chemistry*
  • Ribonucleases / genetics
  • Ribonucleases / metabolism*
  • Substrate Specificity
  • Uridine Monophosphate / chemistry
  • Uridine Monophosphate / metabolism
  • Zebrafish / embryology
  • Zebrafish / microbiology

Substances

  • Bacterial Proteins
  • Poly A
  • Adenosine Monophosphate
  • Uridine Monophosphate
  • Exoribonucleases
  • Ribonucleases
  • exoribonuclease T
  • Adenine

Associated data

  • PDB/4OKE
  • PDB/4OKJ
  • PDB/4OKK