Mitogenic factor secreted by Streptococcus pyogenes is a heat-stable nuclease requiring His122 for activity

Microbiology (Reading). 1997 Jul:143 ( Pt 7):2449-2455. doi: 10.1099/00221287-143-7-2449.

Abstract

The gene encoding a mitogenic factor, termed MF, was cloned from Streptococcus pyogenes and the recombinant MF was overexpressed in Escherichia coli. Both the natural and recombinant MF had heat-resistant nuclease activity. The nuclease activity of MF was characterized using the recombinant protein. MF showed endonuclease activity, digesting ssDNA, dsDNA and tRNA. The optimal pH for the DNase activity of MF was 9.5. The DNase activity was enhanced approximately tenfold by the simultaneous presence of two divalent cations, Mg2+ and Ca2+, compared to either alone and was inhibited by EDTA or NaCl. The heat stability of MF was biphasic; the DNase activity was heat-stable from 0 to 50 degrees C over 80 degrees C but very unstable at around 60 degrees C. DNA digested by MF possessed 5'-phosphorylated and 3'-hydroxylated termini, identical to those obtained by digestion of DNA by pancreatic deoxyribonuclease I. A mutant clone revealed that His122 was a residue essential to the nuclease activity.

MeSH terms

  • Amino Acid Sequence
  • Bacterial Proteins / genetics*
  • Bacterial Proteins / metabolism
  • Deoxyribonucleases / genetics*
  • Deoxyribonucleases / metabolism
  • Enzyme Activation / genetics
  • Histidine / genetics
  • Hot Temperature
  • Mitogens / genetics*
  • Mitogens / metabolism
  • Molecular Sequence Data
  • Recombinant Proteins / genetics
  • Recombinant Proteins / metabolism
  • Sequence Alignment
  • Streptococcus pyogenes / genetics*
  • Streptococcus pyogenes / metabolism

Substances

  • Bacterial Proteins
  • Mitogens
  • Recombinant Proteins
  • Histidine
  • Deoxyribonucleases

Associated data

  • GENBANK/D13428
  • GENBANK/X27241