FtsZDr, a tubulin homologue in radioresistant bacterium Deinococcus radiodurans is characterized as a GTPase exhibiting polymerization/depolymerization dynamics in vitro and FtsZ ring formation in vivo

Modi, Kruti; Tewari, R.; Misra, Hari S.

doi:10.1016/j.biocel.2014.01.015

Cited by 16 publications

(50 citation statements)

References 42 publications

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“…For purification of recombinant proteins, Dr-FtsZ expressing plasmid was constructed as described earlier [20] . Ec-FtsZ expressing plasmid is as described in [25] .…”

Section: Methodsmentioning

confidence: 99%

“…PCR products were cloned at Xho I and Eco RI sites in vector pBAD/HisA under araBAD promoter to yield the pBADEftsA and pBADDftsA plasmids. For in trans expression of FtsZ in FtsA expressing cells, Dr-FtsZ was expressed as FtsZ-GFP on pFTSZGFP [20] . PCR amplified coding sequence of gfp was also cloned in pVHS559 at ScaI and SalI sites to give pVHSgfp.…”

Section: Methodsmentioning

confidence: 99%

“…Dr-FtsZ was purified from recombinant E. coli BL21 (DE3) pLysS as described earlier [20] . Ec-FtsZ was purified from E. coli cells expressing recombinant protein using protocols as described in [27] .…”

Section: Methodsmentioning

confidence: 99%

“…Transcriptome analysis shows that the levels of transcripts for the majority of the divisome components do not change during post irradiation recovery (PIR) [19] . Immunoblotting using antibodies against FtsZ from D. radiodurans (Dr-FtsZ) also shows that the levels of Dr-FtsZ do not change during PIR [20] . Dr-FtsZ is shown to be a slow GTPase that shows polymerization/depolymerization dynamics in vitro and localizes to division sites in D. radiodurans cells [20] .…”

Section: Introductionmentioning

confidence: 99%

“…Immunoblotting using antibodies against FtsZ from D. radiodurans (Dr-FtsZ) also shows that the levels of Dr-FtsZ do not change during PIR [20] . Dr-FtsZ is shown to be a slow GTPase that shows polymerization/depolymerization dynamics in vitro and localizes to division sites in D. radiodurans cells [20] . We observed that E. coli cells overexpressing Dr-FtsA or Dr-FtsZ get elongated, which could have been attributed due to cell division inhibition.…”

Section: Introductionmentioning

confidence: 99%

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Dr-FtsA, an Actin Homologue in Deinococcus radiodurans Differentially Affects Dr-FtsZ and Ec-FtsZ Functions In Vitro

Modi

Misra

2014

PLoS ONE

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The Deinococcus radiodurans genome encodes homologues of divisome proteins including FtsZ and FtsA. FtsZ of this bacterium (Dr-FtsZ) has been recently characterized. In this paper, we study FtsA of D. radiodurans (Dr-FtsA) and its involvement in regulation of FtsZ function. Recombinant Dr-FtsA showed neither ATPase nor GTPase activity and its polymerization was ATP dependent. Interestingly, we observed that Dr-FtsA, when compared with E. coli FtsA (Ec-FtsA), has lower affinity for both Dr-FtsZ and Ec-FtsZ. Also, Dr-FtsA showed differential effects on GTPase activity and sedimentation characteristics of Dr-FtsZ and Ec-FtsZ. For instance, Dr-FtsA stimulated GTPase activity of Dr-FtsZ while GTPase activity of Ec-FtsZ was reduced in the presence of Dr-FtsA. Stimulation of GTPase activity of Dr-FtsZ by Dr-FtsA resulted in depolymerization of Dr-FtsZ. Dr-FtsA effects on GTPase activity and polymerization/depolymerisation characteristics of Dr-FtsZ did not change significantly in the presence of ATP. Recombinant E. coli expressing Dr-FtsA showed cell division inhibition in spite of in trans expression of Dr-FtsZ in these cells. These results suggested that Dr-FtsA, although it lacks ATPase activity, is still functional and differentially affects Dr-FtsZ and Ec-FtsZ function in vitro.

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“…For purification of recombinant proteins, Dr-FtsZ expressing plasmid was constructed as described earlier [20] . Ec-FtsZ expressing plasmid is as described in [25] .…”

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%