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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, �scar; Romero, Manuel; Heeb, Stephan; Espinosa-Urgel, Manuel; C�mara, Miguel; Ramos-Gonz�lez, Mar�a Isabel

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


Authors

�scar Huertas-Rosales

Manuel Romero

Manuel Espinosa-Urgel

Miguel C�mara

Mar�a Isabel Ramos-Gonz�lez



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.

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

Journal Article Type Article
Acceptance Date Jun 23, 2017
Online Publication Date Jul 21, 2017
Deposit Date Jun 26, 2017
Publicly Available Date Jul 21, 2017
Journal Environmental Microbiology
Print ISSN 1462-2912
Electronic ISSN 1462-2920
Publisher Wiley
Peer Reviewed Peer Reviewed
Volume 19
Issue 9
DOI https://doi.org/10.1111/1462-2920.13848
Public URL https://nottingham-repository.worktribe.com/output/873417
Publisher URL http://onlinelibrary.wiley.com/doi/10.1111/1462-2920.13848/full
Contract Date Jun 26, 2017

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