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MatP local enrichment delays segregation independently of tetramer formation and septal anchoring in Vibrio cholerae

ABSTRACT: Vibrio cholerae harbours a primary chromosome derived from the monochromosomal ancestor of the Vibrionales (ChrI) and a secondary chromosome derived from a megaplasmid (ChrII). Both carry a single origin of replication with replication terminating in a diametrically opposite zone, TerI and TerII, respectively. The choreography of TerI and TerII segregation determines when and where cell division occurs. Using a combination of deep sequencing and fluorescence microscopy techniques, we show that TerI and TerII both carry a very high density of MatP binding sites, are both structured into macrodomains, and are both targeted to mid-cell during replication by MatP, demonstrating that ChrII behaves as a bona fide chromosome. We further show that MatP-mediated cohesion and cellular positioning are independent of cell division and tetramer formation, but are linked to the local density of matS sites, in apparent contradiction to the mechanism of action of MatP in Escherichia coli.

ORGANISM(S): Vibrio cholerae

PROVIDER: GSE273189 | GEO | 2024/10/21

REPOSITORIES: GEO

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MatP local enrichment delays segregation independently of tetramer formation and septal anchoring in Vibrio cholerae [3C]

Project description:Vibrio cholerae harbours a primary chromosome derived from the monochromosomal ancestor of the Vibrionales (ChrI) and a secondary chromosome derived from a megaplasmid (ChrII). Both carry a single origin of replication with replication terminating in a diametrically opposite zone, TerI and TerII, respectively. The choreography of TerI and TerII segregation determines when and where cell division occurs. Using a combination of deep sequencing and fluorescence microscopy techniques, we show that TerI and TerII both carry a very high density of MatP binding sites, are both structured into macrodomains, and are both targeted to mid-cell during replication by MatP, demonstrating that ChrII behaves as a bona fide chromosome. We further show that MatP-mediated cohesion and cellular positioning are independent of cell division and tetramer formation, but are linked to the local density of matS sites, in apparent contradiction to the mechanism of action of MatP in Escherichia coli.

2024-10-21 | GSE273188 | GEO

MatP local enrichment delays segregation independently of tetramer formation and septal anchoring in Vibrio cholerae [HiSC2]

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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.

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

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Project description:As in Eukaryotes, bacterial genomes are not randomly folded. Bacterial genetic information is generally carried on a circular chromosome with a single origin of replication from which two replication forks proceed bidirectionaly towards the opposite terminus region. Here we investigate the higher-order architecture of the Escherichia coli genome, showing its partition into two structuraly distinct entities by a complex and intertwined network of contacts: the replication terminus (ter) region and the rest of the chromosome. Outside ter, the condensin MukBEF and the ubiquitous nucleoid-associated protein (NAP) HU promote DNA contacts in the megabase range. Within ter, the MatP protein prevents MukBEF activity and contacts are restricted to ~280 kb creating a domain with distinct structural properties. We also show how other NAPs contribute to nucleoid organization, such as H-NS that restricts short-range interactions. Combined, these results reveal the contributions of major, evolutionary conserved proteins in a bacterial chromosome organization.

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