Macrodomain organization of the Escherichia coli chromosome

We have explored the Escherichia coli chromosome architecture by genetic dissection, using a site‐specific recombination system that reveals the spatial proximity of distant DNA sites and records interactions. By analysing the percentages of recombination between pairs of sites scattered over the chromosome, we observed that DNA interactions were restricted to within subregions of the chromosome. The results indicated an organization into a ring composed of four macrodomains and two less‐structured regions. Two of the macrodomains defined by recombination efficiency are similar to the Ter and Ori macrodomains observed by FISH. Two newly characterized macrodomains flank the Ter macrodomain and two less‐structured regions flank the Ori macrodomain. Also the interactions between sister chromatids are rare, suggesting that chromosome segregation quickly follows replication. These results reveal structural features that may be important for chromosome dynamics during the cell cycle.

[1]  O. Espéli,et al.  Temporal regulation of topoisomerase IV activity in E. coli. , 2003, Molecular cell.

[2]  A. Grossman,et al.  Movement of replicating DNA through a stationary replisome. , 2000, Molecular cell.

[3]  R. Deboy,et al.  Tn7 transposition as a probe of cis interactions between widely separated (190 kilobases apart) DNA sites in the Escherichia coli chromosome , 1996, Journal of bacteriology.

[4]  H. Niki,et al.  Dynamic organization of chromosomal DNA in Escherichia coli. , 2000, Genes & development.

[5]  D. Sherratt,et al.  Spatial and temporal organization of replicating Escherichia coli chromosomes , 2003, Molecular microbiology.

[6]  J. Rebollo,et al.  Detection and possible role of two large nondivisible zones on the Escherichia coli chromosome. , 1988, Proceedings of the National Academy of Sciences of the United States of America.

[7]  J. Errington,et al.  A large dispersed chromosomal region required for chromosome segregation in sporulating cells of Bacillus subtilis , 2002, The EMBO journal.

[8]  A. Segall,et al.  Rearrangement of the bacterial chromosome: forbidden inversions. , 1988, Science.

[9]  N. Cozzarelli,et al.  Linear ordering and dynamic segregation of the bacterial chromosome. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[10]  H. Nash Site-Specific Recombination : Integration , Excision , Resolution , and Inversion of Defined DNA Segments , 1999 .

[11]  A. Grossman,et al.  Chromosome arrangement within a bacterium , 1998, Current Biology.

[12]  N. Cozzarelli,et al.  The Bacterial Condensin MukBEF Compacts DNA into a Repetitive, Stable Structure , 2004, Science.

[13]  H. Niki,et al.  migS, a cis‐acting site that affects bipolar positioning of oriC on the Escherichia coli chromosome , 2004, The EMBO journal.

[14]  J. Errington,et al.  Use of asymmetric cell division and spoIIIE mutants to probe chromosome orientation and organization in Bacillus subtilis , 1998, Molecular microbiology.

[15]  Tom Misteli,et al.  Chromosome positioning in the interphase nucleus. , 2002, Trends in cell biology.

[16]  Stuart Austin,et al.  The segregation of the Escherichia coli origin and terminus of replication , 2002, Molecular microbiology.

[17]  S. Hiraga,et al.  Sister chromosome cohesion of Escherichia coli , 2001, Molecular microbiology.

[18]  A. Segall,et al.  Unequal access of chromosomal regions to each other in Salmonella: probing chromosome structure with phage λ integrase‐mediated long‐range rearrangements , 2004, Molecular microbiology.

[19]  B. Peter,et al.  The topological mechanism of phage lambda integrase. , 1999, Journal of molecular biology.

[20]  A. Grossman,et al.  The extrusion-capture model for chromosome partitioning in bacteria. , 2001, Genes & development.

[21]  Patrick T. McGrath,et al.  Rapid and sequential movement of individual chromosomal loci to specific subcellular locations during bacterial DNA replication. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[22]  A. Landy,et al.  λ Integrase and the λ Int Family , 2002 .

[23]  A. Segall,et al.  Unequal access of chromosomal regions to each other in Salmonella: probing chromosome structure with phage λ integrase‐mediated long‐range rearrangements , 2004 .

[24]  R. Losick,et al.  RacA, a Bacterial Protein That Anchors Chromosomes to the Cell Poles , 2002, Science.

[25]  M. Rossignol,et al.  NKBOR, a mini-Tn10-based transposon for random insertion in the chromosome of Gram-negative bacteria and the rapid recovery of sequences flanking the insertion sites in Escherichia coli. , 2001, Research in microbiology.

[26]  A. Grossman,et al.  Effects of the Chromosome Partitioning Protein Spo0J (ParB) on oriC Positioning and Replication Initiation in Bacillus subtilis , 2003, Journal of bacteriology.

[27]  T. Åkerlund,et al.  Analysis of cell size and DNA content in exponentially growing and stationary-phase batch cultures of Escherichia coli , 1995, Journal of bacteriology.

[28]  N. Higgins DNA Supercoiling and Its Consequences for Chromosome Structure and Function , 1999 .

[29]  J. Roth,et al.  Surveying a supercoil domain by using the gamma delta resolution system in Salmonella typhimurium , 1996, Journal of bacteriology.

[30]  David J Sherratt,et al.  Bacterial Chromosome Dynamics , 2003, Science.

[31]  C. D. Hardy,et al.  Topological domain structure of the Escherichia coli chromosome. , 2004, Genes & development.

[32]  L. Moulin,et al.  Transcription attenuation associated with bacterial repetitive extragenic BIME elements. , 2001, Journal of molecular biology.

[33]  F. Cornet,et al.  Polarization of the Escherichia coli chromosome. A view from the terminus. , 2001, Biochimie.

[34]  R. Stein,et al.  Transcription-induced barriers to supercoil diffusion in the Salmonella typhimurium chromosome. , 2004, Proceedings of the National Academy of Sciences of the United States of America.