A novel system of bacterial cell division arrest implicated in horizontal transmission of an integrative and conjugative element

English version

Autoři: Sotaro Takano ^aff001; Kohei Fukuda ^aff002; Akiko Koto ^aff001; Ryo Miyazaki ^aff001
Působiště autorů: Bioproduction Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), Tsukuba, Japan ^aff001; Department of Bioscience, Graduate School of Science and Technology, Shizuoka University, Hamamatsu, Japan ^aff002; Computational Bio Big Data Open Innovation Laboratory (CBBD-OIL), AIST, Tsukuba, Japan ^aff003; Faculty of Life and Environmental Sciences, University of Tsukuba, Tsukuba, Japan ^aff004
Vyšlo v časopise: A novel system of bacterial cell division arrest implicated in horizontal transmission of an integrative and conjugative element. PLoS Genet 15(10): e32767. doi:10.1371/journal.pgen.1008445
Kategorie: Research Article
doi: https://doi.org/10.1371/journal.pgen.1008445

Souhrn

Integrative and conjugative elements (ICEs) are widespread mobile DNA elements in the prokaryotic world. ICEs are usually retained within the bacterial chromosome, but can be excised and transferred from a donor to a new recipient cell, even of another species. Horizontal transmission of ICEclc, a prevalent ICE in proteobacteria, only occurs from developed specialized transfer competent (tc) cells in the donor population. tc cells become entirely dedicated to the ICE transmission at the cost of cell proliferation. The cell growth impairment is mediated by two ICEclc located genes, parA and shi, but the mechanistic and dynamic details of this process are unknown. To better understand the function of ParA and Shi, we followed their intracellular behavior from fluorescent protein fusions, and studied host cell division at single-cell level. Superresolution imaging revealed that ParA-mCherry colocalized with the host nucleoid while Shi-GFP was enriched at the membrane during the growth impairment. Despite being enriched at different cellular locations, the two proteins showed in vivo interactions, and mutations in the Walker A motif of ParA dislocalized both ParA and Shi. In addition, ParA mutations in the ATPase motif abolished the growth arrest on the host cell. Time-lapse microscopy revealed that ParA and Shi initially delay cell division, suggesting an extension of the S phase of cells, but eventually completely inhibit cell elongation. The parA-shi locus is highly conserved in other ICEclc-related elements, and expressing ParA-Shi from ICEclc in other proteobacterial species caused similar growth arrest, suggesting that the system functions similarly across hosts. The results of our study provide mechanistic insight into the novel and unique system on ICEs and help to understand such epistatic interaction between ICE genes and host physiology that entails efficient horizontal gene transfer.

Klíčová slova:

Adenosine triphosphatase – Cell cycle and cell division – Cell growth – DNA-binding proteins – Fluorescence imaging – Phylogenetic analysis – Pseudomonas putida – Horizontal gene transfer

Zdroje

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