Origin binding activity of the Mycobacterial plasmid pAL5000 replication protein RepB is stimulated through interactions with host factors and coupled expression of repA.

Journal of Bacteriology
Abhijit BasuSujoy K Das Gupta

Abstract

The minimal replication region of the mycobacterial plasmid pAL5000 encompasses the replication origin (ori) and two tandemly organized replication genes, repA and repB, the functions of which are not clearly known. It was observed that when the repA and repB genes were expressed in Escherichia coli, a strong ori binding activity was generated in the host cells. Inactivation of repB led to a complete loss of activity, whereas inactivation of repA had a partial effect, indicating that while repB plays an important role in the process, its activity is stimulated through coexpression of repA. However, this stimulatory effect could be demonstrated only when expression of repA and that of repB were coupled. At a relatively high concentration (1,000 nM), the purified RepB protein was found to form an ori complex with low specificity, which was sensitive to high salt concentrations and challenge by a nonspecific competitor. In contrast, the complex formed by an extract of repA-repB-expressing cells was highly specific and was resistant to both types of challenges. At a 10-fold-lower concentration, RepB did not exhibit ori binding activity, but it could nevertheless form a salt-resistant ori complex in vitro, provided that host factors...Continue Reading

References

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Citations

May 18, 2013·Current Microbiology·Tímea SpakováPeter Pristas
Mar 9, 2004·Plasmid·Nathalie CorneauGisèle LaPointe
Oct 2, 2007·Journal of Bacteriology·Sujoy ChatterjeeSujoy K Das Gupta
Nov 9, 2011·Environmental Microbiology·Yingfei MaBrian Palenik
Feb 6, 2008·FEMS Microbiology Letters·Bidisha BhattacharyaSujoy K Das Gupta
Jan 30, 2008·Microbiology·Twishasri Das GuptaSujoy K Das Gupta

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