Identification and functional annotation of mycobacterial septum formation genes using cell division mutants of Escherichia coli

Sharma, Sujata S. Gaiwala; Kishore, Vimal; Raghunand, Tirumalai R.

doi:10.1016/j.resmic.2015.10.008

Cited by 3 publications

(2 citation statements)

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“…; https://doi.org/10.1101/2023.03. 20.533423 doi: bioRxiv preprint Since no Min like system has been reported in mycobacteria so far, we set out to identify such MinD/ParA/SojA family members, to gain insights into the process of mycobacterial septum site determination, using a target-based genetic complementation approach [19]. Following the identification of MSMEG_3743 and Rv1708 as homologues of Ec MinD, we proceeded to carry out a detailed characterization of the M. smegmatis MinD homologue, MSMEG_3743 and investigated its role in mycobacterial cell division using genetic, biochemical, cell-biological, and comparative structural analysis approaches.…”

Section: Introductionmentioning

confidence: 99%

Septum site placement inMycobacteria- Identification and Characterization of mycobacterial homologues ofEscherichia coliMinD

Kishore

Sharma

Raghunand

2023

Preprint

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A major virulence trait of Mycobacterium tuberculosis (M. tb) is its ability to enter a dormant state within its human host. Since cell division is intimately linked to metabolic shut down, understanding the mechanism of septum formation and its integration with other events in the division pathway is likely to offer clues to the molecular basis of dormancy. The M. tb genome lacks obvious homologues of several conserved cell division proteins, and this study aimed at identifying and functionally characterising mycobacterial homologues of the E.coli septum site specification protein MinD (Ec MinD). Sequence homology based analyses suggested that the genomes of both M. tb and the saprophyte Mycobacterium smegmatis (M. smegmatis) encode two putative <Ec MinD homologues - Rv1708/MSMEG_3743 and Rv3660c/MSMEG_6171. Both Rv1708 and MSMEG_3743 were observed to fully complement the mini-cell phenotype of the E.coli deltaminDE mutant HL1, but the other homologues only partially complemented the mutant phenotype. Over-expression of MSMEG_3743 but not MSMEG_6171 in M. smegmatis led to cell elongation and a drastic decrease in CFU counts, indicating the essentiality of MSMEG_3743 in cell-division. Sequence analysis of MSMEG_3743 showed a conserved Walker A motif, the functional role of which was confirmed by a radiolabelled ATPase activity assay. Rv1708 was observed to interact with the chromosome associated proteins ScpA and ParB, pointing to a link between its septum formation role and chromosome segregation. Comparative structural analyses showed Rv1708 to be closer in similarity to Ec MinD than Rv3660c. In summary we have demonstrated that Rv1708 and MSMEG_3743 are true mycobacterial homologues of Ec MinD, adding a critical missing piece to the mycobacterial cell division puzzle.

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Section: Introductionmentioning

confidence: 99%

Septum site placement inMycobacteria- Identification and Characterization of mycobacterial homologues ofEscherichia coliMinD

Kishore

Sharma

Raghunand

2023

Preprint

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“…Two models explain the asymmetric nature of mycobacterial division -both of which invoke the faster growth of daughter cells containing the old pole, and differ only in their description of whether asymmetry arises due to faster growth of these cells before or after cytokinesis [16][17][18]. Since no Min like system has been reported in mycobacteria so far, we set out to identify such MinD/ParA/SojA family members, to gain insights into the process of mycobacterial septum site determination, using a target-based genetic complementation approach [19]. Following the identification of MSMEG_3743 and Rv1708 as true homologues of Ec MinD, we proceeded to carry out a detailed characterization of the M. smegmatis MinD homologue, MSMEG_3743 and investigated its role in mycobacterial cell division using genetic, biochemical, cell-biological, and comparative structural analysis approaches.…”

Section: Introductionmentioning

confidence: 99%

Septum site placement in Mycobacteria – identification and characterisation of mycobacterial homologues of Escherichia coli MinD

2023

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An Escherichia coli expression model reveals the species-specific function of FtsA from Neisseria gonorrhoeae in cell division

Zou

Ekanayake

et al. 2017

FEMS Microbiology Letters

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Escherichia coli (Ec) has been used to study the function of cell division proteins from different microorganisms, especially when genetic tools are limited for studying these proteins in their native hosts. The expression of ftsA from Neisseria gonorrhoeae (Ng) disrupted cell division in E. coli resulting in a significant increase in cell length. In some cells, FtsANg localised to the division site and the poles of E. coli cells, but the majority of cells showed no specifical localisation. FtsANg did not complement an E. coli ftsA mutant strain. Bacterial two-hybrid and GST pull-down assays indicated that FtsANg interacted with FtsNEc, but no other cell division proteins from E. coli. This interaction was mediated through the 2A and 2B subdomains of FtsANg. This evidence suggests that the function of FtsANg is species specific.

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Identification and functional annotation of mycobacterial septum formation genes using cell division mutants of Escherichia coli

Cited by 3 publications

References 28 publications

Septum site placement inMycobacteria- Identification and Characterization of mycobacterial homologues ofEscherichia coliMinD

Septum site placement inMycobacteria- Identification and Characterization of mycobacterial homologues ofEscherichia coliMinD

Septum site placement in Mycobacteria – identification and characterisation of mycobacterial homologues of Escherichia coli MinD

An Escherichia coli expression model reveals the species-specific function of FtsA from Neisseria gonorrhoeae in cell division

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