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Centromere interactions promote the maintenance of the multipartite genome in Agrobacterium tumefaciens

ABSTRACT: Centromere interactions promote the maintenance of the multipartite genome in Agrobacterium tumefaciens

PROVIDER: PRJNA804267 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR17929500_1.fastq.gz	Fastqsanger.gz
	SRR17929500_2.fastq.gz	Fastqsanger.gz
	SRR17929501_1.fastq.gz	Fastqsanger.gz
	SRR17929501_2.fastq.gz	Fastqsanger.gz
	SRR17929502_1.fastq.gz	Fastqsanger.gz

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Centromere interactions promote the maintenance of the multipartite genome in Agrobacterium tumefaciens

Project description:Many pathogens and symbionts contain multipartite genomes, but how they are stably maintained is poorly understood. The plant pathogen, Agrobacterium tumefaciens, contains four replicons. Recent work indicates that their replication origins cluster at cell poles in a manner that depends on their ParB-family centromeric proteins. Here, we provide evidence that centromeric clustering is mediated by interactions between these centromeric proteins. We further show that the disruption of centromere clustering results in the loss of replicons. Our data establish the role of centromeric clustering in multipartite genome maintenance.

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Project description:Bacterial species from diverse phyla contain multiple replicons, yet how these multipartite genomes are organized and segregated during the cell cycle remains poorly understood. Agrobacterium tumefaciens has a 2.8 Mb circular chromosome (Ch1), a 2.1 Mb linear chromosome (Ch2), and two large plasmids (pAt and pTi). We used this alpha proteobacterium as a model to investigate the global organization and temporal segregation of a multipartite genome. Using Chromosome Conformation Capture (Hi-C) assays, we demonstrate that both the circular and the linear chromosomes but neither of the plasmids have their left and right arms juxtaposed from their origins to their termini, generating inter-arm interactions that require the broadly conserved structural maintenance of chromosomes (SMC) complex. Moreover, our studies revealed two types of inter-replicon interactions: “ori-ori clustering” in which the replication origins of all four replicons interact, and “Ch1-Ch2 alignment” in which the arms of Ch1 and Ch2 interact linearly along their lengths. We show that the centromeric proteins (ParB1 for Ch1 and RepBCh2 for Ch2) are required for both types of inter-replicon contacts. Finally, using fluorescence microscopy, we validated the clustering of the origins and observed their frequent colocalization during segregation. Altogether, our findings provide a high-resolution view of the conformation of a multipartite genome. We hypothesize that inter-centromeric contacts promote the organization and maintenance of diverse replicons.

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