Genomic organization and in vivo characterization of proteolytic activity of FtsH of Mycobacterium smegmatis SN2

Anilkumar, Gopalakrishnapillai and Srinivasan, Ramanujam and Ajitkumar, Parthasarathi (2004) Genomic organization and in vivo characterization of proteolytic activity of FtsH of Mycobacterium smegmatis SN2. In: Microbiology, 150 (8). pp. 2629-2639.

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Abstract

The ftsH gene of Mycobacterium smegmatis SN2 (MsftsH) was cloned from two independent partial genomic DNA libraries and characterized, along with the identification of ephA and folE as the neighbouring upstream and downstream genes respectively. The genomic organization of the MsftsH locus was found to be identical to that of the Mycobacterium tuberculosis ftsH gene (MtftsH) and similar to that of other bacterial genera, but with divergence in the upstream region. The MsftsH gene is 2·3 kb in size and encodes the AAA (ATPases Associated with diverse cellular Activities) family $Zn^2+$-metalloprotease FtsH $(M_sF_t_sH)$ of 85 kDa molecular mass. This was demonstrated from the expression of the full-length recombinant gene in Escherichia coli JM109 cells and from the identification of native MsFtsH in M. smegmatis SN2 cell lysates by Western blotting with anti-MtFtsH and anti- $E_cF_t_sH$ antibodies respectively. The recombinant and the native $M_sF_t_sH$ proteins were found localized to the membrane of E. coli and M. smegmatis cells respectively. Expression of $M_sF_t_sH$ protein in E. coli was toxic and resulted in growth arrest and filamentation of cells. The $M_sf_t_sH$ gene did not complement lethality of a \delta ftsH3 : : kan mutation in E. coli, but when expressed in E. coli cells, it efficiently degraded conventional FtsH substrates, namely \sigma 32 protein and the protein translocase subunit SecY, of E. coli cells

Item Type:	Journal Article
Publication:	Microbiology
Publisher:	Society for General Microbiology
Additional Information:	The copyright of this article belongs to Society for General Microbiology
Department/Centre:	Division of Biological Sciences > Microbiology & Cell Biology
Date Deposited:	07 Dec 2004
Last Modified:	19 Sep 2010 04:17
URI:	http://eprints.iisc.ac.in/id/eprint/2250

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