Mechanistic insights into GTP-dependence and kinetic polarity of FtsZ filament assembly

Chakraborty, Joyeeta; Poddar, Sakshi Mahesh; Dutta, Soumyajit; Bahulekar, Vaishnavi; Harne, Shrikant; Srinivasan, R.; Gayathri, P.

doi:10.1101/2022.10.13.512043

Cited by 2 publications

(1 citation statement)

References 83 publications

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“…SmftsZ gene was PCR amplified from the chromosomal DNA of Spiroplasma melliferum KC3 followed by restriction digestion and ligation at the Nde1 and BamH1 sites into a (His) 6 tag containing pHis17 vector as reported earlier (68). Restriction free cloning (69) was used for cloning of untagged SmFtsZ construct using specific primers.…”

Section: Cloning and Protein Purificationmentioning

confidence: 99%

An amphipathic helix facilitates direct membrane binding of Mycoplasma FtsZ

Dutta,

Poddar,

Chakraborty

et al. 2023

Preprint

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Cell division in bacteria is initiated by constriction of the Z-ring comprising two essential proteins FtsZ and FtsA. Despite our knowledge about the crucial function of the Z-ring in bacterial division, the precise roles and mechanism of how FtsZ and FtsA drive cell constriction remain elusive. FtsZ/FtsA in wall-less bacteria like mycoplasmas is an ideal model system for obtaining mechanistic insights into Z-ring constriction in the absence of cell wall machinery. In this study, we have analyzed FtsZ and FtsA sequences of 113 mycoplasma species and compared with the corresponding protein sequences in cell-walled bacteria. We report a phylogenetically distinct group of 12 species that possess FtsZs without the canonical FtsA interacting conserved C-terminal peptide (CCTP) motif. Interestingly, these FtsZs contain a putative membrane-binding amphipathic helix as an N-terminal or C-terminal extension to the globular FtsZ domain. As a proof-of-concept, we experimentally show that the proposed C-terminal amphipathic helix in M. genitalium FtsZ binds liposomes in vitro as well as localizes to E. coli membrane in vivo. Additionally, we identify a putative cholesterol recognition motif within the C-terminal amphipathic helix region of M. genitalium FtsZ. Our study catalogues the functional variations of membrane attachment by the FtsZ and FtsA system in cell wall-less mycoplasmas and provides a new perspective to study novel functions of FtsZ/A system in cell division.

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Section: Cloning and Protein Purificationmentioning

confidence: 99%

An amphipathic helix facilitates direct membrane binding of Mycoplasma FtsZ

Dutta,

Poddar,

Chakraborty

et al. 2023

Preprint

Self Cite

View full text Add to dashboard Cite

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FtsZ of wall-less bacteria forms ring-like structures

Kasai,

Tahara,

Miyata

et al. 2024

Preprint

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The FtsZ protein is involved in bacterial cell division. In cell-walled bacteria, such asBacillus subtilis, FtsZ forms a ring-like structure, called the Z ring, at the cell division site and acts as a scaffold for cell wall synthesis. The inhibition of cell wall synthesis inB. subtilishas been shown to interfere with the function of the Z ring, causing a loss in cell division control.Spiroplasma, a cell wall-less bacterium, lacks most of the genes involved in cell division; however, theftsZgene remains conserved. The function ofSpiroplasma eriocheirisFtsZ (SeFtsZ) remains to be determined. In the present study, we analyzed the biochemical characteristics of SeFtsZ. Purified SeFtsZ demonstrated lower polymerization capacity and GTPase activity than FtsZ fromE. coliandB. subtilis. We also investigated the relationship between SeFtsZ and SeSepF, which anchors FtsZ to the cell membrane, and found that SeSepF did not contribute to the stability of FtsZ filaments, unlike theB. subtilisSepF. SeFtsZ and SeSepF were produced inE. coliL-forms, where cell wall synthesis was inhibited. SeFtsZ formed ring-like structures in cell wall-lessE. colicells, suggesting that SeFtsZ forms Z rings and is involved in cell division independently of cell wall synthesis.

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Mechanistic insights into GTP-dependence and kinetic polarity of FtsZ filament assembly

Cited by 2 publications

References 83 publications

An amphipathic helix facilitates direct membrane binding of Mycoplasma FtsZ

An amphipathic helix facilitates direct membrane binding of Mycoplasma FtsZ

FtsZ of wall-less bacteria forms ring-like structures

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