Mycobacterium tuberculosis FtsZ requires at least one arginine residue at the C-terminal end for polymerization in vitro

Acta Biochim Biophys Sin (Shanghai). 2010 Jan;42(1):58-69. doi: 10.1093/abbs/gmp105.

Abstract

We examined whether C-terminal residues of soluble recombinant FtsZ of Mycobacterium tuberculosis (MtFtsZ) have any role in MtFtsZ polymerization in vitro. MtFtsZ-deltaC1, which lacks C-terminal extreme Arg residue (underlined in the C-terminal extreme stretch of 13 residues, DDDDVDVPPFMRR), but retaining the penultimate Arg residue (DDDDVDVPPFMR), polymerizes like full-length MtFtsZ in vitro. However, MtFtsZ-deltaC2 that lacks both the Arg residues at the C-terminus (DDDDVDVPPFM), neither polymerizes at pH 6.5 nor forms even single- or double-stranded filaments at pH 7.7 in the presence of 10 mM CaCl(2). Neither replacement of the penultimate Arg residue, in the C-terminal Arg deletion mutant DDDDVDVPPFMR, with Lys or His or Ala or Asp (DDDDVDVPPFMK/H/A/D) enabled polymerization. Although MtFtsZ-deltaC2 showed secondary and tertiary structural changes, which might have affected polymerization, GTPase activity of MtFtsZ-deltaC2 was comparable to that of MtFtsZ. These data suggest that MtFtsZ requires an Arg residue as the extreme C-terminal residue for polymerization in vitro. The polypeptide segment containing C-terminal 67 residues, whose coordinates were absent from MtFtsZ crystal structure, was modeled on tubulin and MtFtsZ dimers. Possibilities for the influence of the C-terminal Arg residues on the stability of the dimer and thereby on MtFtsZ polymerization have been discussed.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alcohol Oxidoreductases / metabolism*
  • Amino Acid Sequence / genetics
  • Arginine / chemistry
  • Arginine / metabolism*
  • Aspartic Acid / metabolism*
  • Bacterial Proteins / chemistry
  • Bacterial Proteins / metabolism*
  • Cloning, Molecular
  • Cytoskeletal Proteins / chemistry
  • Cytoskeletal Proteins / metabolism*
  • DNA-Binding Proteins / metabolism*
  • Gene Expression Regulation, Bacterial
  • Membrane Transport Proteins / metabolism
  • Molecular Sequence Data
  • Mycobacterium tuberculosis / chemistry*
  • Substrate Specificity

Substances

  • Bacterial Proteins
  • Cytoskeletal Proteins
  • DNA-Binding Proteins
  • FtsZ protein, Bacteria
  • Membrane Transport Proteins
  • Aspartic Acid
  • Arginine
  • Alcohol Oxidoreductases
  • C-terminal binding protein