Essential protein SepF of mycobacteria interacts with FtsZ and MurG to regulate cell growth and division

Gupta, Shamba; Banerjee, Srijon Kaushik; Chatterjee, Ayan; Sharma, Arun Kumar; Kundu, Manikuntala; Basu, Joyoti

doi:10.1099/mic.0.000108

Cited by 37 publications

(46 citation statements)

References 46 publications

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“…Consistent with this, B. subtilis cells can still divide, albeit poorly, in the absence of FtsA, and increased levels of SepF can compensate 18 . In Mycobacteria , which naturally lack FtsA, SepF was recently shown to play an essential role in tethering FtsZ to the membrane 19,20 , suggesting it is the sole tether.…”

Section: Assembly Of the Proto-ringmentioning

confidence: 99%

Splitsville: structural and functional insights into the dynamic bacterial Z ring

Haeusser

Margolin²

2016

Nat Rev Microbiol

306

283

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Preface Bacteria must divide in order to increase in number and colonize their niche. Binary fission is the most widespread means of bacterial cell division, but even this relatively simple mechanism displays many variations on a theme. In most bacteria, the tubulin homolog FtsZ assembles into a ring structure (Z ring) at the site of cytokinesis and recruits additional proteins to form a large protein machine (divisome) that spans the membrane. Here we discuss current insights into the regulation of Z ring assembly and how the divisome drives membrane invagination and septal cell wall growth while still flexibly responding to various cellular inputs.

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Section: Assembly Of the Proto-ringmentioning

confidence: 99%

Splitsville: structural and functional insights into the dynamic bacterial Z ring

Haeusser

Margolin²

2016

Nat Rev Microbiol

306

283

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“…Based on this synteny, we named Sven1734 SepF and the additional Streptomyces SepF-like proteins SepF2 (Sven5776) and SepF3 (Sven1372). Sequence alignments showed that all three Streptomyces SepF homologs have the conserved C-terminal domain found in canonical SepF proteins, including the residues essential for protein dimerization and interaction with FtsZ (20,24,33) (Fig. S5A).…”

Section: Resultsmentioning

confidence: 99%

“…Instead, in other bacterial systems FtsZ filaments are tethered to the membrane through interaction with membrane-anchoring proteins such as FtsA, ZipA, and SepF (20)(21)(22)(23)(24). Additional factors, such as the ZapC and ZapD, are critically involved in the stabilization of preformed Z-rings to ensure normal cell division (25)(26)(27).…”

mentioning

confidence: 99%

Two dynamin-like proteins stabilize FtsZ rings during Streptomyces sporulation

Schlimpert

Wasserström

Chandra

et al. 2017

Proc. Natl. Acad. Sci. U.S.A.

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During sporulation, the filamentous bacteria Streptomyces undergo a massive cell division event in which the synthesis of ladders of sporulation septa convert multigenomic hyphae into chains of unigenomic spores. This process requires cytokinetic Z-rings formed by the bacterial tubulin homolog FtsZ, and the stabilization of the newly formed Z-rings is crucial for completion of septum synthesis. Here we show that two dynamin-like proteins, DynA and DynB, play critical roles in this process. Dynamins are a family of large, multidomain GTPases involved in key cellular processes in eukaryotes, including vesicle trafficking and organelle division. Many bacterial genomes encode dynamin-like proteins, but the biological function of these proteins has remained largely enigmatic. Using a cell biological approach, we show that the two Streptomyces dynamins specifically localize to sporulation septa in an FtsZ-dependent manner. Moreover, dynamin mutants have a cell division defect due to the decreased stability of sporulation-specific Z-rings, as demonstrated by kymographs derived from time-lapse images of FtsZ ladder formation. This defect causes the premature disassembly of individual Z-rings, leading to the frequent abortion of septum synthesis, which in turn results in the production of long spore-like compartments with multiple chromosomes. Two-hybrid analysis revealed that the dynamins are part of the cell division machinery and that they mediate their effects on Z-ring stability during developmentally controlled cell division via a network of protein-protein interactions involving DynA, DynB, FtsZ, SepF, SepF2, and the FtsZ-positioning protein SsgB.he active organization and remodeling of cellular membranes is a fundamental process for all organisms. Many of these remodeling events involve the reshaping, fission, or fusion of lipid bilayers to generate new organelles, release transport vesicles, or stabilize specific membrane structures. In eukaryotes, key cellular processes, such as the fission and fusion of mitochondria, the division of chloroplasts, endocytosis, and viral resistance, are mediated by members of the dynamin superfamily (1). Dynamins and dynamin-like proteins are mechanochemical GTPases that polymerize into helical scaffolds at the surface of membranes. GTP hydrolysis is coupled to a radical conformational change in the protein structure that forces the underlying lipid layer into an energetically unstable conformation that promotes membrane rearrangements, probably via a hemifusion intermediate (2, 3).Dynamin-like proteins are found in many bacterial species, yet the precise roles of these proteins are still largely unknown. They share a conserved domain architecture with the canonical human Dynamin 1, including the N-terminal GTPase domain, a neck domain involved in dynamin dimerization, and a trunk domain involved in stimulation of GTPase activity (4, 5). In contrast, bacterial dynamins lack the pleckstrin homology motif and proline-rich sequences found in classical dynamins and contain instead o...

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“…Since then, the TetR/Pip‐OFF system has been successfully used by several groups to study essential genes and validate drug targets in vitro and in vivo in different mycobacterial species (Serafini et al ., 2009, 2013; Cortes et al ., 2011; Di Luca et al ., 2012; Mondino et al ., 2013; Ventura et al ., 2013; Ahmed et al ., 2014, 2016; Bazet Lyonnet et al ., 2014; Bhowmick et al ., 2014; Boldrin et al ., 2014; Kolly et al ., 2014a,b; Pandey and Rodriguez, 2014; Verma and Chatterji, 2014; Gola et al ., 2015; Gupta et al ., 2015; Mori et al ., 2015; Hu et al ., 2016; Degiacomi et al ., 2017). This broad experience taught us that the main drawback of this otherwise very succesful system was the strength of the P ptr promoter, which causes overexpression of the gene of interest, when this is physiologically expressed at low level, with resulting accumulation of its product.…”

Section: Introductionmentioning

confidence: 99%

Promoter mutagenesis for fine‐tuning expression of essential genes in Mycobacterium tuberculosis

Boldrin

Degiacomi

Serafini

et al. 2017

Microbial Biotechnology

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SummaryA range of regulated gene expression systems has been developed for mycobacteria in the last few years to facilitate the study of essential genes, validate novel drug targets and evaluate their vulnerability. Among these, the TetR/Pip‐OFF repressible promoter system was successfully used in several mycobacterial species both in vitro and in vivo. In the first version of the system, the repressible promoter was Pptr, a strong Pip‐repressible promoter of Streptomyces pristinaespiralis, which might hamper effective downregulation of genes with a low basal expression level. Here, we report an enhanced system that allows more effective control of genes expressed at low level. To this end, we subjected Pptr to targeted mutagenesis and produced 16 different promoters with different strength. Three of them, weaker than the wild‐type promoter, were selected and characterized showing that they can indeed improve the performances of TetR/Pip‐OFF repressible system both in vitro and in vivo increasing its stringency. Finally, we used these promoters to construct a series of bacterial biosensors with different sensitivity to DprE1 inhibitors and developed a whole‐cell screening assay to identify inhibitors of this enzyme.

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Essential protein SepF of mycobacteria interacts with FtsZ and MurG to regulate cell growth and division

Cited by 37 publications

References 46 publications

Splitsville: structural and functional insights into the dynamic bacterial Z ring

Splitsville: structural and functional insights into the dynamic bacterial Z ring

Two dynamin-like proteins stabilize FtsZ rings during Streptomyces sporulation

Promoter mutagenesis for fine‐tuning expression of essential genes in Mycobacterium tuberculosis

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