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Genome-wide ParB2-DNA and RctB-DNA interaction analyses in Vibrio cholerae

ABSTRACT: Replication and segregation are the two main processes that maintain chromosomes in growing cells. In bacteria, replication and transcription have been proposed to provide motive force in chromosome segregation. Recently, ParB2, a segregation protein, encoded by V. cholerae chromosome II (chrII) was also found to influence chrII replication. V. cholerae chrII replication is primarily controlled by its specific initiator protein, RctB. Here, we have screened for ParB2 and RctB binding sites using a genome-wide DNA binding analysis (ChIP-chip). We report the identification of a new region containing additional RctB and ParB2 binding sites, and suggest the mechanisms to coordinate replication initiation with segregation of chromosomes.

ORGANISM(S): Vibrio cholerae O1 biovar El Tor str. N16961 Vibrio cholerae

PROVIDER: GSE72978 | GEO | 2015/09/14

SECONDARY ACCESSION(S): PRJNA295519

REPOSITORIES: GEO

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Genome-wide ParB2-DNA and RctB-DNA interaction analyses in Vibrio cholerae

Project description:Replication and segregation are the two main processes that maintain chromosomes in growing cells. In bacteria, replication and transcription have been proposed to provide motive force in chromosome segregation. Recently, ParB2, a segregation protein, encoded by V. cholerae chromosome II (chrII) was also found to influence chrII replication. V. cholerae chrII replication is primarily controlled by its specific initiator protein, RctB. Here, we have screened for ParB2 and RctB binding sites using a genome-wide DNA binding analysis (ChIP-chip). We report the identification of a new region containing additional RctB and ParB2 binding sites, and suggest the mechanisms to coordinate replication initiation with segregation of chromosomes. ParB2 binding DNA (Cy5) vs total DNA (input, Cy3) in wildtype (CVC209), RctB binding DNA (Cy5) vs total DNA (input, Cy3) in wildtype (CVC209), RctB binding DNA (Cy5) vs total DNA (input, Cy3) in MCH1 (CVC2099, rctB-deleted strain), Biological replicates: 3 or 2

2015-09-14 | E-GEOD-72978 | biostudies-arrayexpress

Marker Frequency data of genomic loci extracted from N16961 V. cholerae strain grown fro 10h in M9-fructose + 0.2% L-Arabinose, in M9-fructose to exp phase and to stationary phase

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2018-11-13 | GSE114980 | GEO

Chromosome segregation in Streptococcus pneumoniae

Project description:Segregation of replicated chromosomes during cell division is an essential process in all organisms. Chromosome segregation is promoted by the action of the DNA-binding ParB protein in the rod-shaped model bacterium Bacillus subtilis. How oval shaped bacteria, such as the human pathogen Streptococcus pneumoniae, efficiently segregate their chromosomes is poorly understood. Here, we show that the pneumococcal homolog of ParB is enriched at four centromere-like DNA sequences (parS sites) that are present near the origin of replication.

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Kinetochores coordinate pericentromeric cohesion and early DNA replication by Cdc7-Dbf4 kinase recruitment

Project description:Centromeres play several important roles in ensuring proper chromosome segregation. Not only do they promote kinetochore assembly for microtubule attachment, but they also support robust sister chromatid cohesion at pericentromeres and facilitate replication of centromeric DNA early in S phase. However, it is still elusive how centromeres orchestrate all these functions at the same site. Here we show that the budding yeast Dbf4-dependent kinase (DDK) accumulates at kinetochores in telophase, facilitated by the Ctf19 kinetochore complex. This promptly recruits Sld3-Sld7 replication initiator proteins to pericentromeric replication origins so that they initiate replication early in S phase. Furthermore DDK at kinetochores independently recruits the Scc2-Scc4 cohesin loader to centromeres in G1 phase. This enhances cohesin loading and facilitates robust pericentromeric cohesion in S phase. Thus, we have found the central mechanism by which kinetochores orchestrate early S phase DNA replication and robust sister chromatid cohesion at microtubule attachment sites. Measurement of genome replication time for various S. cerevisiae strains. For each strain two samples were analysed: a replicating sample (from S phase) and a non-replicating sample (from G2 phase).

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Kinetochores coordinate pericentromeric cohesion and early DNA replication by Cdc7-Dbf4 kinase recruitment

2013-06-06 | GSE41982 | GEO

Vibrio cholerae

Project description:Vibrio cholerae -chromosome 1

| PRJDB12712 | ENA

Real-time dynamics of Plasmodium Ndc80 reveals unusual modes of chromosome segregation during parasite proliferative stages

Project description:Eukaryotic cell proliferation requires chromosome replication and precise segregation to ensure daughter cells have identical genomic copies. The genus Plasmodium, the causative agent of malaria, has developed remarkable characteristic of nuclear division throughout its lifecycle to meet some peculiar and unique challenges of DNA replication and chromosome segregation. The parasite undergoes atypical endomitosis and endoreduplication with an intact nuclear membrane and intranuclear mitotic spindle. To understand diverse modes of cell division in Plasmodium we have studied the behaviour and composition of a key part of the mitotic apparatus, the outer kinetochore Ndc80 complex that attaches the centromere of chromosomes to the microtubules of the mitotic spindle. Using Ndc80 live-cell imaging in Plasmodium berghei we observe dynamic changes throughout proliferative life-stages including highly unusual kinetochore arrangements in sexual stages of the parasite. Furthermore, we identify a divergent Spc24 component of Ndc80 complex, previously thought to be missing, and completing a canonical, albeit unusual, Ndc80 complex structure. Altogether our studies reveal the kinetochore as an ideal starting point towards understanding non-canonical modes of chromosome segregation and cell division in Plasmodium.

2020-06-09 | PXD017619 | Pride

The nucleoid occlusion factor Noc controls DNA replication initiation in Staphylococcus aureus [WGS-Seq]

Project description:Successive division events in the spherically shaped bacterium Staphylococcus aureus are oriented in three alternating perpendicular planes. The mechanisms that underlie this relatively unique pattern of division and coordinate it with chromosome segregation remain largely unknown. Thus far, the only known spatial regulator of division in this organism is the nucleoid occlusion protein Noc that inhibits assembly of the cytokinetic ring over the chromosome. However, Noc is not essential in S. aureus, indicating that additional regulators are likely to exist. To search for these factors, we screened for mutants that are synthetic lethal with Noc inactivation. Our characterization of these mutants led to the discovery that S. aureus Noc also controls the initiation of DNA replication. We show that cells lacking Noc over-initiate and mutations in the initiator gene dnaA suppress this defect. Importantly, these dnaA mutations also partially suppress the division problems associated with ∆noc. Reciprocally, we show that over-expression of DnaA enhances the over-initiation and cell division phenotypes of cells lacking Noc. Thus, a single factor both blocks cell division over chromosomes and helps to ensure that new rounds of DNA replication are not initiated prematurely. This degree of economy in coordinating key cell biological processes has not been observed in rod-shaped bacteria and may reflect the challenges posed by the reduced cell volume and complicated division pattern of this spherical pathogen.

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