TY - JOUR

T1 - Specialized and shared functions of diguanylate cyclases and phosphodiesterases in Streptomyces development

AU - Haist, Julian

AU - Neumann, Sara Alina

AU - Al-Bassam, Mahmoud M

AU - Lindenberg, Sandra

AU - Elliot, Marie A

AU - Tschowri, Natalia

N1 - Funding information: We thank Andreas Latoscha and Mirka E. Wörmann for comments on the manuscript, Susan Schlimpert for the pSS5 plasmid and Heike Bähre for excellent technical assistance with LC?MS/MS analysis. Research in Natalia Tschowri’s lab is funded by the DFG Emmy Noether Program (TS 325/1?1) and the DFG Priority Program SPP 1879 (TS 325/2?1 and TS 325/2?2), and in Marie Elliot’s lab by the Natural Sciences and Engineering Council of Canada’s Discovery Grant program (RGPIN?2015?04681). Open access funding enabled and organized by Projekt DEAL.

PY - 2020/11

Y1 - 2020/11

N2 - The second messenger bis-3,5-cyclic di-guanosine monophosphate (c-di-GMP) determines when Streptomyces initiate sporulation. c-di-GMP signals are integrated into the genetic differentiation network by the regulator BldD and the sigma factor σWhiG . However, functions of the development-specific diguanylate cyclases (DGCs) CdgB and CdgC, and the c-di-GMP phosphodiesterases (PDEs) RmdA and RmdB, are poorly understood. Here, we provide biochemical evidence that the GGDEF-EAL domain protein RmdB from S. venezuelae is a monofunctional PDE that hydrolyzes c-di-GMP to 5'pGpG. Despite having an equivalent GGDEF-EAL domain arrangement, RmdA cleaves c-di-GMP to GMP and exhibits residual DGC activity. We show that an intact EAL motif is crucial for the in vivo function of both enzymes since strains expressing protein variants with an AAA motif instead of EAL are delayed in development, similar to null mutants. Transcriptome analysis of ∆cdgB, ∆cdgC, ∆rmdA, and ∆rmdB strains revealed that the c-di-GMP specified by these enzymes has a global regulatory role, with about 20% of all S. venezuelae genes being differentially expressed in the cdgC mutant. Our data suggest that the major c-di-GMP-controlled targets determining the timing and mode of sporulation are genes involved in cell division and the production of the hydrophobic sheath that covers Streptomyces aerial hyphae and spores.

AB - The second messenger bis-3,5-cyclic di-guanosine monophosphate (c-di-GMP) determines when Streptomyces initiate sporulation. c-di-GMP signals are integrated into the genetic differentiation network by the regulator BldD and the sigma factor σWhiG . However, functions of the development-specific diguanylate cyclases (DGCs) CdgB and CdgC, and the c-di-GMP phosphodiesterases (PDEs) RmdA and RmdB, are poorly understood. Here, we provide biochemical evidence that the GGDEF-EAL domain protein RmdB from S. venezuelae is a monofunctional PDE that hydrolyzes c-di-GMP to 5'pGpG. Despite having an equivalent GGDEF-EAL domain arrangement, RmdA cleaves c-di-GMP to GMP and exhibits residual DGC activity. We show that an intact EAL motif is crucial for the in vivo function of both enzymes since strains expressing protein variants with an AAA motif instead of EAL are delayed in development, similar to null mutants. Transcriptome analysis of ∆cdgB, ∆cdgC, ∆rmdA, and ∆rmdB strains revealed that the c-di-GMP specified by these enzymes has a global regulatory role, with about 20% of all S. venezuelae genes being differentially expressed in the cdgC mutant. Our data suggest that the major c-di-GMP-controlled targets determining the timing and mode of sporulation are genes involved in cell division and the production of the hydrophobic sheath that covers Streptomyces aerial hyphae and spores.

KW - EAL

KW - GGDEF

KW - Streptomyces

KW - c-di-GMP

KW - diguanylate cyclase

KW - phosphodiesterase

UR - http://www.scopus.com/inward/record.url?scp=85089378063&partnerID=8YFLogxK

U2 - 10.1111/mmi.14581

DO - 10.1111/mmi.14581

M3 - Article

C2 - 32797697

VL - 114

SP - 808

EP - 822

JO - Molecular microbiology

JF - Molecular microbiology

SN - 0950-382X

IS - 5

ER -