RT info:eu-repo/semantics/article
T1 Genetic dissection of the regulatory network associated with high c-di-GMP levels in Pseudomonas putida KT2440
A1 Ramos González, María Isabel
A1 Travieso, María L.
A1 Soriano, María I.
A1 Matilla, Miguel A.
A1 Huertas Rosales, Óscar
A1 Barrientos Moreno, Laura
A1 García Tagua, Víctor
A1 Espinosa Urgel, Manuel
K1 Pseudomonas
K1 Biofilm
K1 c-di-GMP
K1 crinkle morphology
K1 two-component regulatory system
AB Most bacteria grow in nature forming multicellular structures named biofilms. The bacterial second messenger cyclic diguanosine monophosphate (c-di-GMP) is a key player in the regulation of the transition from planktonic to sessile lifestyles and this regulation is crucial in the development of biofilms. In Pseudomonas putida KT2440, Rup4959, a multidomain response regulator with diguanylate cyclase activity, when overexpressed causes an increment in the intracellular levels of c-di-GMP that gives rise to a pleiotropic phenotype consisting of increased biofilm formation and crinkly colony morphology. In a broad genomic screen we have isolated mutant derivatives that lose the crinkly morphology, designed as cfc (crinkle free colony). A total of 19 different genes have been identified as being related with the emergence of the cfc phenotype either because the expression or functionality of Rup4959 is compromised, or due to a lack of transduction of the c-di-GMP signal to downstream elements involved in the acquisition of the phenotype. Discernment between these possibilities was investigated by using a c-di-GMP biosensor and by HPLC-MS quantification of the second messenger. Interestingly five of the identified genes encode proteins with AAA+ ATPase domain. Among the bacterial determinants found in this screen are the global transcriptional regulators GacA, AlgU and FleQ and two enzymes involved in the arginine biosynthesis pathway. We present evidences that this pathway seems to be an important element to both the availability of the free pool of the second messenger c-di-GMP and to its further transduction as a signal for biosynthesis of biopolimers. In addition we have identified an uncharacterized hybrid sensor histidine kinase whose phosphoaceptor conserved histidine residue has been shown in this work to be required for in vivo activation of the orphan response regulator Rup4959, which suggests these two elements constitute a two-component phosphorelay system.
SN 1664-302X
YR 2016
FD 2016
LK http://riull.ull.es/xmlui/handle/915/35627
UL http://riull.ull.es/xmlui/handle/915/35627
LA en
DS Repositorio institucional de la Universidad de La Laguna
RD 18-nov-2024