Overexpression of the response regulator rpaA causes an impaired cell division in the Cyanobacterium Synechocystis sp. PCC 6803

J Gen Appl Microbiol. 2020 Jun 17;66(2):121-128. doi: 10.2323/jgam.2020.01.004. Epub 2020 Mar 13.

Abstract

In photosynthetic microorganisms, cell cycle progression depends on day and night cycles; however, how cell division is regulated in response to these environmental changes is poorly understood. RpaA has been implicated in the signal output from both circadian clocks and light/dark conditions in the unicellular spherical-celled cyanobacterium Synechocystis sp. PCC 6803. In the present study, we investigated the involvement of a two-component response regulator RpaA in cell division regulation. Firstly, we examined the effects of rpaA overexpression on cell morphology and the expression levels of cell division genes. We observed an increase in the volume of non-dividing cells and a high proportion of dividing cells in rpaA-overexpressing strains by light microscopy. The expression levels of selected cell division-related genes were higher in the rpaA-overexpressing strain than in the wild type, including minD of the Min system; cdv3 and zipN, which encode two divisome components; and murB, murC, and pbp2, which are involved in peptidoglycan (PG) synthesis. Moreover, in the rpaA-overexpressing strain, the outer membrane and cell wall PG layer were not smooth, and the outer membrane was not clearly visible by transmission electron microscopy. These results demonstrated that rpaA overexpression causes an impaired cell division, which is accompanied by transcriptional activation of cell division genes and morphological changes in the PG layer and outer membrane.

Keywords: Synechocystis; cell division; circadian rhythm; cyanobacteria; cytokinesis; transcription factor.

MeSH terms

  • Bacterial Outer Membrane / ultrastructure*
  • Cell Cycle Proteins / genetics*
  • Cell Cycle Proteins / metabolism
  • Cell Division*
  • Circadian Clocks / genetics*
  • Gene Expression Regulation, Bacterial
  • Microscopy
  • Microscopy, Electron, Transmission
  • Peptidoglycan / biosynthesis
  • Real-Time Polymerase Chain Reaction
  • Synechocystis / cytology
  • Synechocystis / genetics*
  • Synechocystis / ultrastructure*

Substances

  • Cell Cycle Proteins
  • Peptidoglycan