Molecular interactions and their predictive roles in cell pole determination in bacteria

Chaudhary, Reema; Mishra, Shruti; Kota, Swathi; Misra, Hari S.

doi:10.1080/1040841x.2020.1857686

Cited by 5 publications

(10 citation statements)

References 228 publications

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“…The divIVA is shown to be an essential gene for the normal growth of this bacterium, and its chromosomal copy could be replaced with selection marker only when drDivIVA was expressed episomally (Chaudhary et al, 2021). The effect of phosphorylation on drDivIVA functions was further evaluated by an allelic replacement of mutant copy with the wild type copy in the chromosome of this bacterium.…”

Section: Resultsmentioning

confidence: 99%

“…The replacement of phosphorylation site of 201 st threonine with alanine in DivIVA has resulted into an elongated and bulged cell phenotype in S. pneumoniae (Fleurie et al, 2012). Further, the important roles of DivIVA in the determination of polarity and planes of cell division have been shown at least in cocci (Chaudhary et al, 2021). DivIVA in general localises at the negative curvature, which is also marked by cardiolipin microdomains at least in B. subtilis (Renner and Weibel, 2011).…”

Section: Discussionmentioning

confidence: 99%

“…D. radiodurans lacks cardiolipin microdomains but its DivIVA (i) undergoes phosphorylation by RqkA at T19 site that maps in proximity with the polar determining motif, (ii) contains an extended CTD when compared with DivIVA of other bacteria, (iii) is an essential protein and its CTD domain contributes to the positioning of septum during normal growth in this bacterium (Chaudhary et al, 2021), and (iv) interacts with cognate cell division and genome segregation proteins (Maurya et al, 2018). Therefore, the localization of DivIVA and its phosphomutants, and their dynamics with respect to FtsZ ring positioning and genome segregation was monitored.…”

Section: Discussionmentioning

confidence: 99%

“…General procedures of fluorescence microscopy using confocal microscope (Model Olympus Cell IX 83 Fluoview 3000) were as described earlier (Chaudhary et al, 2021). In brief, the bacterial cultures were grown in TYG broth, fixed with 4% paraformaldehyde for 10 min on ice and washed two times with phosphate-buffered saline (pH 7.4).…”

Section: Methodsmentioning

confidence: 99%

“…The transformants were grown for several rounds in the presence of kanamycin (8μg/ml). The cycles of alternate streaking on TYG agar plate and sub-culturing in TYG broth were repeated for several rounds till homogenous replacement of target with insertion sequences has been achieved (Chaudhary. et al, 2021).…”

Section: Generation Of Chromosomal Insertions In Divivamentioning

confidence: 99%

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DivIVA phosphorylation at threonine affects its dynamics and cell cycle in Deinococcus radiodurans

Chaudhary

Kota

Misra

2022

Preprint

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RqkA, a γ radiation responsive Ser/Thr quinoprotein kinase, is characterized for its role in radioresistance in Deinoocccus radiodurans. DivIVA is a cell division protein involved in determination of cell pole and division site in bacteria. RqkA phosphorylated cognate DivIVA (drDivIVA) at Threonine 19 (T19) residue located in its pole recognition motif. The phospho-mimetic replacement of T19 (T19E) functioned differently than phospho-ablative (T19A) and drDivIVA proteins. T19E-RFP expressing in wild type background showed arrest in dynamics of drDivIVA, and loss of interaction with genome segregation protein. divIVA is shown to be an essential gene in this bacterium. The allelic replacement of divIVA with T19E-RFP was not tolerated unless drDivIVA was expressed episomally while there was no effect of this replacement with T19A-RFP and drDivIVA-RFP. These results suggested that the T19 phosphorylation in drDivIVA by RqkA has affected in vivo functions of DivIVA that would render cell cycle arrest in this bacterium.

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Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Generation Of Chromosomal Insertions In Divivamentioning

confidence: 99%

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DivIVA phosphorylation at threonine affects its dynamics and cell cycle in Deinococcus radiodurans

Chaudhary

Kota

Misra

2022

Preprint

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The tri-flow adaptiveness of codes in major evolutionary transitions

Girard

2024

BioSystems

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DivIVA Regulates Its Expression and the Orientation of New Septum Growth in Deinococcus radiodurans

2021

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In rod shaped Gram-negative bacteria, FtsZ localization at mid cell position is regulated by the gradient of MinCDE complex across the poles. In round shaped bacteria, which lack predefined poles, the next plane of cell division is perpendicular to previous plane and the determination of site for FtsZ assembly is still intriguing. Deinococcus radiodurans, a coccus bacterium, is characterized by its extraordinary resistance to DNA damage. DivIVA, a putative component of Min system in this bacterium, interacts with cognate cell division and genome segregation proteins. Here, we report that deletion of chromosomal copy of DivIVA was possible only when wild type copy of DivIVA was expressed in trans on the plasmid. However, the deletion of C-terminal domain of DivIVA (CTD mutant) was possible but produced distinguishable phenotypes like smaller cells, slower growth and tilted septum orientation in D. radiodurans. In trans expression of DivIVA in CTD mutant could restore these features of wild type. Interestingly, the overexpression of DivIVA led to delayed separation of tetrad from octet state in both trans-complemented divIVA mutant and wild type cells. The CTD mutant showed upregulation of yggS-divIVAN operon. Both wild type and CTD mutant formed FtsZ foci, however unlike wild type, the position of foci in the mutant cells was found to be away from conjectural mid-cell position in cocci. Notably, DivIVA-RFP localizes to septum during cell division at the new division site. These results suggested that DivIVA is an essential protein in D. radiodurans and its C-terminal domain plays an important role in the regulation of its expression and orientation of new septal growth in this bacterium. Importance: In rod-shaped Gram-negative bacteria, the mid-cell position for binary fission is relatively easy to model. In cocci that do not have predefined poles, the plane of next cell division is shown to be perpendicular to the previous plane. However, the molecular basis of perpendicularity is not known in cocci. The DivIVA protein of Deinococcus radiodurans, a coccus bacterium, physically interacts with septum and establishes macromolecular interactions with genome segregation proteins through its N-terminal domain and with MinC through C-terminal domain. Here, we have brought forth some evidence to suggest that DivIVA is essential for growth, plays an important role in cell-polarity determination and its C-terminal domain plays a crucial role in the growth of new septum in correct orientation as well as regulation of its expression.

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Molecular interactions and their predictive roles in cell pole determination in bacteria

Cited by 5 publications

References 228 publications

DivIVA phosphorylation at threonine affects its dynamics and cell cycle in Deinococcus radiodurans

DivIVA phosphorylation at threonine affects its dynamics and cell cycle in Deinococcus radiodurans

The tri-flow adaptiveness of codes in major evolutionary transitions

DivIVA Regulates Its Expression and the Orientation of New Septum Growth in Deinococcus radiodurans

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