Functional characterization of multiple pas domain-containing diguanylate cyclases in synechocystis sp. Pcc 6803

Ko Ishikawa, Chihiro Chubachi, Saeko Tochigi, Naomi Hoshi, Seiji Kojima, Mamoru Hyodo, Yoshihiro Hayakawa, Tadaomi Furuta, Kota Kera, Nobuyuki Uozumi

Research output: Contribution to journalArticlepeer-review

Abstract

Bis-(3′–5′)-cyclic dimeric guanosine monophosphate (c-di-GMP) is a second messenger known to control a variety of bacterial processes. The model cyanobacterium, Synechocystis sp. PCC 6803, has a score of genes encoding putative enzymes for c-di-GMP synthesis and degradation. However, most of them have not been functionally characterized. Here, we chose four genes in Synechocystis (dgcA–dgcD), which encode proteins with a GGDEF, diguanylate cyclase (DGC) catalytic domain and multiple Per-ARNT-Sim (PAS) conserved regulatory motifs, for detailed analysis. Purified DgcA, DgcB and DgcC were able to catalyze synthesis of c-di-GMP from two GTPs in vitro. DgcA had the highest activity, compared with DgcB and DgcC. DgcD did not show detectable activity. DgcA activity was specific for GTP and stimulated by the divalent cations, magnesium or manganese. Full activity of DgcA required the presence of the multiple PAS domains, probably because of their role in protein dimerization or stability. Synechocystis mutants carrying single deletions of dgcA–dgcD were not affected in their growth rate or biofilm production during salt stress, suggesting that there was functional redundancy in vivo. In contrast, overexpression of dgcA resulted in increased biofilm formation in the absence of salt stress. In this study, we characterize the enzymatic and physiological function of DgcA–DgcD, and propose that the PAS domains in DgcA function in maintaining the enzyme in its active form.

Original languageEnglish
Pages (from-to)659-668
Number of pages10
JournalMicrobiology (United Kingdom)
Volume166
Issue number7
DOIs
Publication statusPublished - 2020

Keywords

  • C-di-GMP
  • Cyanobacterium
  • Diguanylate cyclase
  • PAS
  • Synechocystis sp. PCC 6803

ASJC Scopus subject areas

  • Microbiology

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