C-shaped cells caused by expression of an ftsA mutation in Escherichia coli

Gayda, Randall C.; Henk, Margaret C.; Leong, D

doi:10.1128/jb.174.16.5362-5370.1992

Cited by 40 publications

(35 citation statements)

References 31 publications

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“…However, the overproduction of FtsA carboxy-terminal truncations causes a peculiar phenotype, in which the cells coil forming C-shaped (short) and coiled (longer) cells (CC phenotype). This phenotype has already been observed by Gayda et al (17) in wild-type cells containing a form of FtsA lacking the last 28 residues encoded by a plasmid.…”

Section: Resultssupporting

confidence: 76%

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Role of the Carboxy Terminus of Escherichia coli FtsA in Self-Interaction and Cell Division

et al. 2000

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The role of the carboxy terminus of the Escherichia coli cell division protein FtsA in bacterial division has been studied by making a series of short sequential deletions spanning from residue 394 to 420. Deletions as short as 5 residues destroy the biological function of the protein. Residue W415 is essential for the localization of the protein into septal rings. Overexpression of the ftsA alleles harboring these deletions caused a coiled cell phenotype previously described for another carboxy-terminal mutation (Gayda et al., J. Bacteriol. 174:5362-5370, 1992), suggesting that an interaction of FtsA with itself might play a role in its function. The existence of such an interaction was demonstrated using the yeast two-hybrid system and a protein overlay assay. Even these short deletions are sufficient for impairing the interaction of the truncated FtsA forms with the wild-type protein in the yeast two-hybrid system. The existence of additional interactions between FtsA molecules, involving other domains, can be postulated from the interaction properties shown by the FtsA deletion mutant forms, because although unable to interact with the wild-type and with FtsA⌬1, they can interact with themselves and cross-interact with each other. The secondary structures of an extensive deletion, FtsA⌬27, and the wild-type protein are indistinguishable when analyzed by Fourier transform infrared spectroscopy, and moreover, FtsA⌬27 retains the ability to bind ATP. These results indicate that deletion of the carboxy-terminal 27 residues does not alter substantially the structure of the protein and suggest that the loss of biological function of the carboxy-terminal deletion mutants might be related to the modification of their interacting properties.FtsA is an essential cell division protein of Escherichia coli that is widely conserved in bacteria. Together with ftsZ, which codes for a GTPase analog of the eukaryotic tubulin, ftsA forms one of the most frequently conserved gene pairs among the cell division genes in the eubacteria. Based on sequence homology it has been proposed that FtsA belongs to the sugar kinase/hsp70/actin superfamily (4). This superfamily comprises several proteins with a common two-domain topology and the ability to bind and hydrolyze ATP. FtsA binds to columns of ATP-agarose and can be isolated from cells either as a phosphorylated or a nonphosphorylated form (29), but so far no other biochemical function has been described for this protein.FtsA is present both in the cytoplasm and in the cytoplasmic membrane (29), where it forms a structural part of the septum (32). It has been proposed that FtsA is a component of a membrane-associated complex (septator or divisome), which would include periplasmic, transmembrane, and cytoplasmic proteins acting coordinately to perform septation (27,35). Genetic analysis suggests that FtsA may interact, directly or indirectly, with other cell division proteins, such as FtsZ, PBP3, FtsQ, and FtsN (9,10,24,33,34). The FtsZ/FtsA ratio is important for cell division, an...

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Section: Resultssupporting

confidence: 76%

“…Involvement of the protein carboxy terminus in the interaction was suspected from previous results showing that overproduction of a carboxy-terminally truncated form of FtsA causes the formation of long fibers in the cells that also show a curved morphology phenotype (17).…”

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confidence: 99%

Role of the Carboxy Terminus of Escherichia coli FtsA in Self-Interaction and Cell Division

et al. 2000

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“…This differential extension must occur across the entire length of a cell in a regular manner to result in such uniform coils. Similar morphologies were recently observed during the expression of a mutant E. coli ftsA gene (17), suggesting that this phenotype could be caused by an altered interaction between FtsA and FtsZ. The coiling and kinking were also seen within longer filaments (Fig.…”

Section: Rml Sk L F Agv Apt E Asupporting

confidence: 57%

Rhizobium meliloti contains a novel second homolog of the cell division gene ftsZ

Margolin

Long

1994

J Bacteriol

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We have identified a second homolog of the cell division gene,ftsZ, in the endosymbiont Rhizobium meliloti. The ftsZ2 gene was cloned by screening a genomic A library with a probe derived from PCR amplification of a highly conserved domain. It encodes a 36-kDa protein which shares a high level of sequence similarity with the FtsZ proteins of Escherichia coli and BaciUus subtilis and FtsZl (Zl) of R. meliloti but lacks the carboxy-terminal region conserved in other FtsZ proteins. The identity of theftsZ2 gene product was confirmed both by in vitro transcription-translation in an R. meliloti S-30 extract and by overproduction in R. meliloti cells. As with Zl, the overproduction of FtsZ2 in E. coli inhibited cell division and induced filamentation, although to a lesser extent than with Zl. However, the expression offtsZ2 in E. coli under certain conditions caused some cells to coil dramatically, a phenotype not observed during Zl overproduction. Although several Tn3-GUS (glucuronidase) insertions in a plasmid-borneftsZ2 gene failed to cross into the chromosome, one interruption in the chromosomal ftsZ2 gene was isolated, suggesting that ftsZ2 is nonessential for viability. The two ftsZ genes were genetically mapped to the R. meliloti main chromosome, approximately 100 kb apart.The earliest known step in Escherichia coli cell division is the formation of a ring consisting of the FtsZ protein at the division site (5). FtsZ is also necessary for the formation of both vegetative and asymmetric sporulation septa in Bacillus subtilis (2). Cells containing very high levels of FtsZ or a temperature-sensitive FtsZ under nonpermissive conditions will form long, nonseptate filaments, presumably because the division ring cannot be formed (3, 39). It is not known how FtsZ, which is uniformly distributed during most of the cell cycle, assembles at the correct time in the center of the predivisional cell. However, the recent demonstrations that E. coli FtsZ binds GTP and has GTPase activity (10,26,29) suggest that GTP binding and hydrolysis by FtsZ could catalyze the assembly of the ring structure, in a manner analogous to filament formation by tubulins (34). The three previously sequenced FtsZ proteins, from E. coli, B. subtilis, and Rhizobium meliloti (1,21,40), all share a completely conserved 13-amino-acid domain that is very similar to the proposed GTP binding domain of tubulins (10) MATERIALS AND METHODSBacterial strains and plasmids. All strains and plasmids used in this study are listed in Table 1.Bacterial growth and induction conditions. All E. coli strains were grown in Luria broth (LB) (30) plus 10 ,ug of tetracycline and 50 ,ug of ampicillin per ml for plasmids in host strain XL1-Blue. Isopropyl-,B-D-thiogalactopyranoside (IPTG) inductions of E. coli cells were done by streaking colonies on agar plates containing LB, antibiotics, and 1 mM IPTG. R. meliloti cells were induced for protein overproduction by the addition of 1 mM IPTG to cells growing in LB plus 10 ,ug of tetracycline per ml in late exponential phase.Pre...

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“…The genetic analysis of mutants shows a potential nucleoid segregation role for numerous genes. Some of these genes are involved in DNA replication, cell division, structural maintenance of chromosome, or SOS system, or they encode histone-like proteins (1,5,6,9,11,20,22,23,54). Others were associated with the formation of anucleate cells (13,14) or polyploidy (48).…”

Section: Discussionmentioning

confidence: 99%

Cytoplasmic Filament-Deficient Mutant of Treponema denticola Has Pleiotropic Defects

2001

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In Treponema denticola, a ribbon-like structure of cytoplasmic filaments spans the cytoplasm at all stages of the cell division process. Insertional inactivation was used as a first step to determine the function of the cytoplasmic filaments. A suicide plasmid was constructed that contained part of cfpA and a nonpolar erythromycin resistance cassette (ermF and ermAM) inserted near the beginning of the gene. The plasmid was electroporated into T. denticola, and double-crossover recombinants which had the chromosomal copy of cfpA insertionally inactivated were selected. Immunoblotting and electron microscopy confirmed the lack of cytoplasmic filaments. The mutant was further analyzed by dark-field microscopy to determine cell morphology and by the binding of two fluorescent dyes to DNA to assess the distribution of cellular nucleic acids. The cytoplasmic filament protein-deficient mutant exhibited pleiotropic defects, including highly condensed chromosomal DNA, compared to the homogeneous distribution of the DNA throughout the cytoplasm in a wild-type cell. Moreover, chains of cells are formed by the cytoplasmic filament-deficient mutant, and those cells show reduced spreading in agarose, which may be due to the abnormal cell length. The chains of cells and the highly condensed chromosomal DNA suggest that the cytoplasmic filaments may be involved in chromosome structure, segregation, or the cell division process in Treponema.

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C-shaped cells caused by expression of an ftsA mutation in Escherichia coli

Cited by 40 publications

References 31 publications

Role of the Carboxy Terminus of Escherichia coli FtsA in Self-Interaction and Cell Division

Role of the Carboxy Terminus of Escherichia coli FtsA in Self-Interaction and Cell Division

Rhizobium meliloti contains a novel second homolog of the cell division gene ftsZ

Cytoplasmic Filament-Deficient Mutant of Treponema denticola Has Pleiotropic Defects

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