Polymerization of FtsZ, a Bacterial Homolog of Tubulin

Romberg, Laura; Simon, Martha N.; Erickson, Harold P.

doi:10.1074/jbc.m009033200

Cited by 198 publications

(287 citation statements)

References 66 publications

Supporting

Mentioning

259

Contrasting

Unclassified

Order By: Relevance

“…This report deals with the assembly of FtsZ at conditions known to induce lateral association of protofilaments (9,35,36,39). Lateral assembly of FtsZ implies cooperativity, as pointed out recently (22) and is evidenced by the concentration dependence of the D209N and D212E mutants. Linear assembly of FtsZ in a noncooperative fashion was recently described (20)(21)(22).…”

Section: Discussionmentioning

confidence: 79%

GTP Hydrolysis of Cell Division Protein FtsZ: Evidence that the Active Site Is Formed by the Association of Monomers

et al. 2001

View full text Add to dashboard Cite

The essential prokaryotic cell division protein FtsZ is a tubulin homologue that forms a ring at the division site. FtsZ forms polymers in a GTP-dependent manner. Recent biochemical evidence has shown that FtsZ forms multimeric structures in vitro and in vivo and functions as a self-activating GTPase. Structural analysis of FtsZ points to an important role for the highly conserved tubulin-like loop 7 (T7-loop) in the self-activation of GTP hydrolysis. The T7-loop was postulated to form the active site together with the nucleotide-binding site on an adjacent FtsZ monomer. To characterize the role of the T7-loop of Escherichia coli FtsZ, we have mutagenized residues M206, N207, D209, D212, and R214. All the mutant proteins, except the R214 mutant, are severely affected in polymerization and GTP hydrolysis. Charged residues D209 and D212 cannot be substituted with a glutamate residue. All mutants interact with wildtype FtsZ in vitro, indicating that the T7-loop mutations do not abolish FtsZ self-association. Strikingly, in mixtures of wild-type and mutant proteins, most mutants are capable of inhibiting wild-type GTP hydrolysis. We conclude that the T7-loop is part of the active site for GTP hydrolysis, formed by the association of two FtsZ monomers.

show abstract

Section: Discussionmentioning

confidence: 79%

GTP Hydrolysis of Cell Division Protein FtsZ: Evidence that the Active Site Is Formed by the Association of Monomers

et al. 2001

View full text Add to dashboard Cite

show abstract

“…Most methods for monitoring FtsZ assembly indicate that the assembly of protofilaments is COOPERATIVE, with a defined critical concentration for assembly 33,34 . However, the presence of only one binding face for a new monomer to attach to an existing protofilament indicates an ISODESMIC ASSEMBLY mechanism 35,36 , as does the appearance of single protofilaments in electron micrographs of assembled FtsZ in the absence of bundling factors. How can the apparent cooperativity of assembly be rationalized with the microscopic evidence of single-stranded protofilaments?…”

Section: Author Manuscriptmentioning

confidence: 99%

FtsZ and the division of prokaryotic cells and organelles

Margolin

2005

Nat Rev Mol Cell Biol

560

484

View full text Add to dashboard Cite

Binary fission of many prokaryotes as well as some eukaryotic organelles depends on the FtsZ protein, which self-assembles into a membrane-associated ring structure early in the division process. FtsZ is homologous to tubulin, the building block of the microtubule cytoskeleton in eukaryotes. Recent advances in genomics and cell-imaging techniques have paved the way for the remarkable progress in our understanding of fission in bacteria and organelles.Duplication of cells occurs by the division of a mother cell into two daughter cells. This process, known as cytokinesis, provides the force to split cells and is spatially regulated to faithfully partition the genetic material. The cytoskeleton has a crucial role in cytokinesis. In animal and fungal cells, a medial ring of actin and myosin, helped by other proteins, contracts to divide the cell. Plant cells use a cell plate, and are guided by actin filaments and microtubules. Prokaryotes, which include bacteria and archaea, possess homologues of eukaryotic cytoskeletal proteins. Most prokaryotes use a tubulin homologue, a protein known as FtsZ, to divide. Eukaryotic organelles such as chloroplasts and mitochondria evolved from bacteria, and all chloroplasts and some mitochondria use FtsZ to divide.FtsZ is thought to be the first protein to localize to the site of future division in bacteria 1 , and it assembles into what is known as the Z ring (FIG. 1). In Escherichia coli, the Z ring recruits at least ten other proteins, all of which are required for the progression and completion of cytokinesis 2 , as will be discussed below. Whereas the cytokinetic apparatus in eukaryotic cells and even organelles can be observed directly in stained thin sections by transmission electron microscopy, the Z ring and its associated factors are not detectable by these methods. This is probably because the protein machinery is largely membrane bound and resides in a densely populated cytoplasmic environment. However, the development of green fluorescent protein (GFP) fusion and improved immunofluorescence techniques over the past 10 years have been crucial in allowing the first direct visualization of many cellular components and their dynamics. For example, using GFP fusions, the dynamics of the Z ring and other components of the cell-division protein machinery can now be visualized in living bacterial cells. The combination of these new cytological tools with genomics and the Competing interests statementThe author declares no competing financial interests. HHS Public Access Author Manuscript Author ManuscriptAuthor ManuscriptAuthor Manuscript already powerful genetics of several model systems have spurred rapid progress in our understanding of the molecular cell biology of bacteria and eukaryotic organelles. This review will discuss how the recent revolutions in genomics and cell biology have provided new evolutionary and mechanistic insights into how bacterial cells and organelles divide. The FtsZ family of proteins Conservation of FtsZFtsZ is a highly conserved protein ...

show abstract

“…The protein was aliquoted and stored at -80 uC. FtsZ was put through a cycle of calcium-aided assembly and disassembly to select the active and nonaggregated protein (Romberg et al, 2001). All the following experiments were repeated in triplicate.…”

Section: Methodsmentioning

confidence: 99%

“…E. coli FtsZ (EcFtsZ) was overexpressed and purified from the recombinant E. coli BL21 strain carrying pET11b (kindly provided by Professor H. P. Erickson, Duke University) as described previously (Lu et al, 2001;Romberg et al, 2001). The purity of the protein was analysed by 10 % SDS-PAGE and its concentration was determined by the Bradford method (Bradford, 1976).…”

Section: Methodsmentioning

confidence: 99%

“…The purity of the protein was analysed by 10 % SDS-PAGE and its concentration was determined by the Bradford method (Bradford, 1976). The protein was aliquoted and stored at -80 uC.FtsZ was put through a cycle of calcium-aided assembly and disassembly to select the active and nonaggregated protein (Romberg et al, 2001). All the following experiments were repeated in triplicate.…”

mentioning

confidence: 99%

See 1 more Smart Citation

Phenylpropanoids inhibit protofilament formation of Escherichia coli cell division protein FtsZ

Hemaiswarya

Soudaminikkutty

Narasumani

et al. 2011

Journal of Medical Microbiology

View full text Add to dashboard Cite

The earliest step in cell division in bacteria is the assembly of FtsZ, an essential cell division protein, into a ring at the division site. FtsZ has GTPase activity and can assemble in vitro to form protein filaments. The present work involved the study of eight phenylpropanoids (cinnamic, p-coumaric, caffeic, chlorogenic, ferulic, 3,4-dimethoxycinnamic and 2,4,5-trimethoxycinnamic acids and eugenol) as inhibitors of Escherichia coli FtsZ. Phenylpropanoids make up the majority of our diet and act as antibacterial agents. Polymerization and GTPase inhibition assays showed that chlorogenic and caffeic acids were the most active amongst these (IC 50 of 70 and 106 mM, respectively). Circular dichroism studies indicated that chlorogenic acid perturbed the protein conformation and electron microscopy showed distorted filaments. Bacillus subtilis 168 cells treated with the phenylpropanoids were longer when compared to the control. The highest binding energy was observed between chlorogenic acid and the homology modelled E. coli FtsZ, which was consistent with the experimental results. A strong negative correlation was observed between binding energy and inhibition of the polymerization ability. 3D-Quantitative structureactivity relationship studies using GTPase activity indicated that the presence of more hydrophilic groups around the 39-and 49-carbon increased the activity. The effect of stress-induced formation of cell filamentation has to be understood before confirming the role of phenylpropanoids as FtsZ inhibitors. INTRODUCTIONAntimicrobial resistance is a global concern due to its increased prevalence. The major issues are the reduced susceptibility to currently available antimicrobial agents and shortage of newly approved compounds. A growing number of Gram-positive (Staphylococcus aureus, Streptococcus pneumoniae, etc.) and Gram-negative (Pseudomonas aeruginosa, Klebsiella pneumoniae, Escherichia coli, etc.) pathogens are responsible for infection in both the community and health-care settings (Furtado & Nicolau, 2010). Around 19-60 % of Staphylococcus aureus isolates from ocular infections have been shown to be resistant to macrolides, penicillin and fluoroquinolones (McDonald & Blondeau, 2010). The overall levels of antibiotic resistance can be dramatically decreased by focusing on the overuse of antibiotics, the pharmacokinetic and pharmacodynamic properties of different drug formulations, and use of proper hygiene and protective barriers. There is also a need for the introduction of new antimicrobial agents with novel targets.FtsZ is a bacterial cytoskeleton protein which assembles into a protofilament in a GTP-dependent manner. This forms a dynamic Z-ring at the mid-cell position. A study done in E. coli showed that 12 different proteins congregate at the Z-ring in a sequentially dependent pathway, causing assembly of the septal ring that guides the synthesis of the circumferential septum (Buddelmeijer & Beckwith, 2002;Bernhardt & de Boer, 2003;Romberg & Levin, 2003; Schmidt et al., 2004). The septal r...

show abstract

Polymerization of FtsZ, a Bacterial Homolog of Tubulin

Cited by 198 publications

References 66 publications

GTP Hydrolysis of Cell Division Protein FtsZ: Evidence that the Active Site Is Formed by the Association of Monomers

GTP Hydrolysis of Cell Division Protein FtsZ: Evidence that the Active Site Is Formed by the Association of Monomers

FtsZ and the division of prokaryotic cells and organelles

Phenylpropanoids inhibit protofilament formation of Escherichia coli cell division protein FtsZ

Contact Info

Product

Resources

About