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The Pseudomonas putida CsrA/RsmA homologues negatively affect c-di-GMP pools and biofilm formation through the GGDEF/EAL response regulator CfcR

Huertas-Rosales, Óscar; Romero, Manuel; Heeb, Stephan; Espinosa-Urgel, Manuel; Cámara, Miguel; Ramos-González, María Isabel

Authors

Óscar Huertas-Rosales

Manuel Romero

Stephan Heeb

Manuel Espinosa-Urgel

Miguel Cámara

María Isabel Ramos-González



Abstract

Expression of cfcR, encoding the only GGDEF/EAL response regulator in Pseudomonas putida, is transcriptionally regulated by RpoS, ANR and FleQ, and the functionality of CfcR as a diguanylate cyclase requires the multisensor CHASE3/GAF hybrid histidine kinase named CfcA. Here an additional level of cfcR control, operating post-transcriptionally via the RNA-binding proteins RsmA, RsmE and RsmI, is unraveled. Specific binding of the three proteins to an Rsm binding motif (5’CANGGANG3’) encompassing the translational start codon of cfcR was confirmed. Although RsmA exhibited the highest binding affinity to the cfcR transcript, single deletions of rsmA, rsmE or rsmI, caused minor derepression in CfcR translation compared to a ∆rsmIEA triple mutant. RsmA also showed a negative impact on c-di-GMP levels in a double mutant ∆rsmIE through the control of cfcR, which is responsible for most of the free c-di-GMP during stationary phase in static conditions. In addition, a CfcR-dependent c-di-GMP boost was observed during this stage in ∆rsmIEA confirming the negative effect of Rsm proteins on CfcR translation and explaining the increased biofilm formation in this mutant compared to the wild type. Overall these results suggest that CfcR is a key player in biofilm formation regulation by the Rsm proteins in P. putida.

Journal Article Type Article
Journal Environmental Microbiology
Print ISSN 1462-2912
Electronic ISSN 1462-2920
Publisher Wiley
Peer Reviewed Peer Reviewed
Volume 19
Issue 9
APA6 Citation Huertas-Rosales, Ó., Romero, M., Heeb, S., Espinosa-Urgel, M., Cámara, M., & Ramos-González, M. I. (in press). The Pseudomonas putida CsrA/RsmA homologues negatively affect c-di-GMP pools and biofilm formation through the GGDEF/EAL response regulator CfcR. Environmental Microbiology, 19(9), https://doi.org/10.1111/1462-2920.13848
DOI https://doi.org/10.1111/1462-2920.13848
Publisher URL http://onlinelibrary.wiley.com/doi/10.1111/1462-2920.13848/full
Copyright Statement Copyright information regarding this work can be found at the following address: http://creativecommons.org/licenses/by/4.0

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Copyright Statement
Copyright information regarding this work can be found at the following address: http://creativecommons.org/licenses/by/4.0





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