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Sensory domain of the cell cycle kinase CckA in Caulobacter crescentus regulates the differential DNA binding activity of the master regulator CtrA

Sharath Narayanan, Lokesh Kumar, View ORCID ProfileSunish Kumar Radhakrishnan
doi: https://doi.org/10.1101/300178
Sharath Narayanan
School of Biology, Indian Institute of Science Education and Research, Thiruvananthapuram 695016, Kerala, India.
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Lokesh Kumar
School of Biology, Indian Institute of Science Education and Research, Thiruvananthapuram 695016, Kerala, India.
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Sunish Kumar Radhakrishnan
School of Biology, Indian Institute of Science Education and Research, Thiruvananthapuram 695016, Kerala, India.
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  • ORCID record for Sunish Kumar Radhakrishnan
  • For correspondence: sunish@iisertvm.ac.in
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Abstract

Sophisticated signaling mechanisms allow bacterial cells to cope with environmental and intracellular challenges. Activation of specific pathways facilitates the cells to overcome cellular damage and thereby warrant integrity. Here we demonstrate the pliability of the CckA-CtrA two component signaling system in the freshwater bacterium, Caulobacter crescentus. Our forward genetic screen to analyse suppressor mutations that can negate the chromosome segregation block induced by the topoisomerase IV inhibitor, NstA, yielded various point mutations in the cell cycle histidine kinase, CckA. Notably, we identified a point mutation in the PAS-B domain of CckA, which resulted in increased levels of phosphorylated CtrA (CtrA∼P), the master cell cycle regulator. Surprisingly, this increase in CtrA∼P levels did not translate into a genome-wide increase in the DNA occupancy of CtrA, but specifically enriched its affinity to the chromosomal origin of replication, Cori, and a very small sub-set of CtrA regulated promoters. We show that through this enhanced binding of CtrA to the Cori, cells are able to overcome the toxic defects rendered by stable NstA through a possible slow down in the chromosome cycle. Taken together, our work opens up an unexplored and intriguing aspect of the CckA-CtrA signal transduction pathway. The distinctive DNA binding nature of CtrA and its regulation by CckA might also be crucial for pathogenesis because of the highly conserved nature of CckA-CtrA pathway in alphaproteobacteria.

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Posted April 14, 2018.
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Sensory domain of the cell cycle kinase CckA in Caulobacter crescentus regulates the differential DNA binding activity of the master regulator CtrA
Sharath Narayanan, Lokesh Kumar, Sunish Kumar Radhakrishnan
bioRxiv 300178; doi: https://doi.org/10.1101/300178
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Sensory domain of the cell cycle kinase CckA in Caulobacter crescentus regulates the differential DNA binding activity of the master regulator CtrA
Sharath Narayanan, Lokesh Kumar, Sunish Kumar Radhakrishnan
bioRxiv 300178; doi: https://doi.org/10.1101/300178

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