DNA replication initiation is required for mid‐cell positioning of FtsZ rings in Caulobacter crescentus

Polymerization of the GTPase FtsZ to form a structure called the Z‐ring is the earliest known step in bac‐terial cell division. Mid‐cell Z‐ring assembly coincides with the beginning of the replication cycle in the differentiating bacterium Caulobacter crescentus. Z‐ring disassembly occurs at the end of the division cycle, resulting in the complete degradation of FtsZ from both stalked and swarmer progeny cells. New Z‐rings can only form in the replicative stalked cell. Conditional mutants in DNA replication were used to determine what role DNA replication events play in the process of Z‐ring assembly at different stages in the cell cycle. Z‐ring assembly occurred even when early stages of DNA replication were blocked; however, the Z‐rings were localized at a subpolar region of the cell. Z‐rings only assembled at the proper mid‐cell location if DNA replication had initiated. Z‐ring assembly coincided with areas containing little or no DNA, and Z‐rings could not form over an unreplicated chromosome. Overexpressed FtsZ in the absence of DNA replication did not stimulate productive mid‐cell Z‐ring assembly but, instead, caused the ends of cells to constrict over an extended area away from the nucleoid. These results indicate that the state of chromosome replication is a major determinant of Z‐ring localization in Caulobacter.

[1]  M. Osley,et al.  Circular organization of the DNA synthetic pathway in Caulobacter crescentus , 1982, Journal of bacteriology.

[2]  J. Lutkenhaus,et al.  The MinC component of the division site selection system in Escherichia coli interacts with FtsZ to prevent polymerization. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[3]  A. Kornberg,et al.  Duplex opening by dnaA protein at novel sequences in initiation of replication at the origin of the E. coli chromosome , 1988, Cell.

[4]  Y. Brun,et al.  Cell cycle regulation and cell type-specific localization of the FtsZ division initiation protein in Caulobacter. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[5]  W. Donachie,et al.  Transcription of essential cell division genes is linked to chromosome replication in Escherichia coli , 2001, Molecular microbiology.

[6]  E. Harry.  Coordinating DNA replication with cell division: lessons from outgrowing spores. , 2001, Biochimie.

[7]  W. Margolin,et al.  FtsZ ring clusters in min and partition mutants: role of both the Min system and the nucleoid in regulating FtsZ ring localization , 1999, Molecular microbiology.

[8]  J. Errington,et al.  Selection of the midcell division site in Bacillus subtilis through MinD‐dependent polar localization and activation of MinC , 1999, Molecular microbiology.

[9]  Y. Brun,et al.  CtrA mediates a DNA replication checkpoint that prevents cell division in Caulobacter crescentus , 2000, The EMBO journal.

[10]  R. B. Jensen,et al.  The Caulobacter crescentus smc gene is required for cell cycle progression and chromosome segregation. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[11]  M. Marinus,et al.  Correlation of DNA adenine methylase activity with spontaneous mutability in Escherichia coli K-12. , 1984, Gene.

[12]  Y. Brun,et al.  Cell cycle-dependent transcriptional and proteolytic regulation of FtsZ in Caulobacter. , 1998, Genes & development.

[13]  A. Kornberg,et al.  Purified dnaA protein in initiation of replication at the Escherichia coli chromosomal origin of replication. , 1983, Proceedings of the National Academy of Sciences of the United States of America.

[14]  Arthur Kornberg,et al.  The dnaA protein complex with the E. coli chromosomal replication origin (oriC) and other DNA sites , 1984, Cell.

[15]  B. Ely,et al.  Use of pulsed field gel electrophoresis and transposon mutagenesis to estimate the minimal number of genes required for motility in Caulobacter crescentus. , 1989, Genetics.

[16]  E. Bi,et al.  Interaction between the min locus and ftsZ , 1990, Journal of bacteriology.

[17]  L Shapiro,et al.  Rate, origin, and bidirectionality of Caulobacter chromosome replication as determined by pulsed-field gel electrophoresis. , 1989, Proceedings of the National Academy of Sciences of the United States of America.

[18]  Y. Brun,et al.  Dominant C‐terminal deletions of FtsZ that affect its ability to localize in Caulobacter and its interaction with FtsA , 1998, Molecular microbiology.

[19]  N Nanninga,et al.  Role of the nucleoid in the toporegulation of division. , 1990, Research in microbiology.

[20]  W. Margolin,et al.  Influence of the Nucleoid on Placement of FtsZ and MinE Rings in Escherichia coli , 2001, Journal of bacteriology.

[21]  L. Shapiro,et al.  A Caulobacter DNA methyltransferase that functions only in the predivisional cell. , 1994, Journal of molecular biology.

[22]  A. Tormo,et al.  The ftsA gene product: a possible connection between DNA replication and septation in Escherichia coli. , 1985, Journal of general microbiology.

[23]  M. O’Donnell,et al.  Replisome assembly at oriC, the replication origin of E. coli, reveals an explanation for initiation sites outside an origin. , 1999, Molecular cell.

[24]  R. Losick,et al.  Inactivation of FtsI inhibits constriction of the FtsZ cytokinetic ring and delays the assembly of FtsZ rings at potential division sites. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[25]  E. Boye,et al.  The role of dam methyltransferase in the control of DNA replication in E. coli , 1990, Cell.

[26]  J. Gober,et al.  Cell Cycle–Dependent Polar Localization of Chromosome Partitioning Proteins in Caulobacter crescentus , 1997, Cell.

[27]  Y. Bum,et al.  The Dimorphic Life Cycle of Caulobacter and Stalked Bacteria , 2000 .

[28]  E. Harry,et al.  Mid‐cell Z ring assembly in the absence of entry into the elongation phase of the round of replication in bacteria: co‐ordinating chromosome replication with cell division , 2000, Molecular microbiology.

[29]  R. B. Jensen,et al.  A moving DNA replication factory in Caulobacter crescentus , 2001, The EMBO journal.

[30]  N. Nanninga,et al.  Timing of FtsZ Assembly in Escherichia coli , 1999, Journal of bacteriology.

[31]  J. Plumbridge.  The role of dam methylation in controlling gene expression. , 1987, Biochimie.

[32]  N. Agabian,et al.  Envelope-associated nucleoid from Caulobacter crescentus stalked and swarmer cells , 1977, Journal of bacteriology.

[33]  L. Shapiro,et al.  Caulobacter Lon protease has a critical role in cell-cycle control of DNA methylation. , 1996, Genes & development.

[34]  M. Osley,et al.  Regulation of flagellin synthesis in the cell cycle of Caulobacter: Dependence on DNA replication , 1977, Cell.

[35]  Ian T. Paulsen,et al.  Complete genome sequence of Caulobacter crescentus , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[36]  J. Gober,et al.  The chromosome partitioning protein, ParB, is required for cytokinesis in Caulobacter crescentus , 2001, Molecular microbiology.

[37]  Lucy Shapiro,et al.  Cell Type-Specific Phosphorylation and Proteolysis of a Transcriptional Regulator Controls the G1-to-S Transition in a Bacterial Cell Cycle , 1997, Cell.

[38]  E. Harry,et al.  Co‐ordinating DNA replication with cell division in bacteria: a link between the early stages of a round of replication and mid‐cell Z ring assembly , 1999, Molecular microbiology.

[39]  K. Nordström,et al.  FtsZ ring formation without subsequent cell division after replication runout in Escherichia coli , 2000, Molecular Microbiology.

[40]  E. Boye,et al.  Expression of the Escherichia coli dam gene , 1992, Molecular microbiology.

[41]  A. Newton,et al.  Signal Transduction and Cell Cycle Checkpoints in Developmental Regulation of Caulobacter , 2000 .

[42]  M. Osley,et al.  Mutational analysis of developmental control in Caulobacter crescentus. , 1977, Proceedings of the National Academy of Sciences of the United States of America.

[43]  Lucy Shapiro,et al.  Cell Cycle Control by an Essential Bacterial Two-Component Signal Transduction Protein , 1996, Cell.

[44]  A. Newton,et al.  Cloning and cell cycle-dependent expression of DNA replication gene dnaC from Caulobacter crescentus , 1990, Journal of bacteriology.

[45]  L. Shapiro,et al.  Expression of Caulobacter dnaA as a function of the cell cycle , 1994, Journal of bacteriology.

[46]  G. Marczynski.  Chromosome Methylation and Measurement of Faithful, Once and Only Once per Cell Cycle Chromosome Replication inCaulobacter crescentus , 1999, Journal of bacteriology.

[47]  L. Shapiro,et al.  A cell cycle-regulated bacterial DNA methyltransferase is essential for viability. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[48]  A. Goffeau,et al.  The complete genome sequence of the Gram-positive bacterium Bacillus subtilis , 1997, Nature.

[49]  C. Woldringh,et al.  Actively replicating nucleoids influence positioning of division sites in Escherichia coli filaments forming cells lacking DNA , 1989, Journal of bacteriology.

[50]  G. Marczynski,et al.  Physiological consequences of blocked Caulobacter crescentus dnaA expression, an essential DNA replication gene , 2001, Molecular microbiology.

[51]  J. Errington,et al.  Postseptational chromosome partitioning in bacteria. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[52]  G. Churchward,et al.  Control of cyclic chromosome replication in Escherichia coli. , 1991, Microbiological reviews.

[53]  L. Shapiro,et al.  Requirement of the carboxyl terminus of a bacterial chemoreceptor for its targeted proteolysis. , 1993, Science.

[54]  J. Lutkenhaus,et al.  Unique and overlapping roles for ZipA and FtsA in septal ring assembly in Escherichia coli , 2002, The EMBO journal.

[55]  M. W. Woude,et al.  Evidence for global regulatory control of pilus expression in Escherichia coli by Lrp and DNA methylation: model building based on analysis of pap , 1992, Molecular microbiology.

[56]  Y. Brun,et al.  Cell cycle and positional constraints on FtsZ localization and the initiation of cell division in Caulobacter crescentus , 2001, Molecular microbiology.

[57]  P. Modrich,et al.  Escherichia coli K-12 clones that overproduce dam methylase are hypermutable , 1981, Journal of bacteriology.

[58]  A. Grossman,et al.  Localization of bacterial DNA polymerase: evidence for a factory model of replication. , 1998, Science.

[59]  Lucy Shapiro,et al.  Genes directly controlled by CtrA, a master regulator of the Caulobacter cell cycle , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[60]  E. Bi,et al.  FtsZ ring formation in fts mutants , 1996, Journal of bacteriology.

[61]  J. Poindexter.  BIOLOGICAL PROPERTIES AND CLASSIFICATION OF THE CAULOBACTER GROUP , 1964, Bacteriological reviews.

[62]  S. Moriya,et al.  Regulation of initiation of the chromosomal replication by DnaA‐boxes in the origin region of the Bacillus subtilis chromosome. , 1988, The EMBO journal.

[63]  J. Lutkenhaus,et al.  Inhibition of FtsZ polymerization by SulA, an inhibitor of septation in Escherichia coli. , 1998, Proceedings of the National Academy of Sciences of the United States of America.