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Related Experiment Videos

McrBs, a modulator peptide for McrBC activity

D Panne1, E A Raleigh, T A Bickle

  • 1Department of Microbiology, Biozentrum, Basel University, Klingelbergstrasse 70, CH-4056 Basel, Switzerland.

The EMBO Journal
|September 16, 1998
PubMed
Summary

The McrBC enzyme complex from E. coli restricts methylated DNA. Its activity is modulated by McrBs, which inhibits or enhances DNA cleavage depending on McrC levels.

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Area of Science:

  • Molecular Biology
  • Enzymology
  • Bacteriology

Background:

  • McrBC is a methylation-dependent endonuclease in E. coli K-12.
  • It recognizes and restricts DNA with specific methylated cytosine patterns.
  • The enzyme complex involves McrBL, McrBs, and McrC gene products.

Purpose of the Study:

  • To elucidate the role of the McrBs protein in McrBC endonuclease activity.
  • To understand the subunit stoichiometry required for DNA cleavage.
  • To investigate the regulatory mechanism of McrBC by McrBs.

Main Methods:

  • Purification of McrBs protein.
  • In vitro analysis of DNA cleavage activity.
  • Investigating the effect of McrBs on McrBC in varying ratios.

Main Results:

  • DNA cleavage requires McrBL, McrC, GTP, and Mg2+.
  • Optimal cleavage occurs at a 3-5:1 ratio of McrBL to McrC.
  • McrBs inhibits cleavage at optimal ratios by sequestering McrC, but enhances it when McrC is in excess.

Conclusions:

  • McrBs acts as a modulator of McrBC activity.
  • Its function is dependent on the relative concentrations of McrBL and McrC.
  • McrBs likely regulates McrBC by binding to McrC in a GTP-dependent manner.

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