connecting quorum sensing, c-di-gmp, pel polysaccharide, and biofilm formation in pseudomonas aeruginosa through tyrosine phosphatase tpba (pa3885)连接群体感应,c-di-gmp贝利多糖,在铜绿假单胞菌生物膜的形成通过酪氨酸磷酸酶tpba(pa3885).pdfVIP

Connecting Quorum Sensing, c-di-GMP, Pel Polysaccharide, and Biofilm Formation in Pseudomonas aeruginosa through Tyrosine Phosphatase TpbA (PA3885) 1 1,2,3 Akihiro Ueda , Thomas K. Wood * 1 Artie McFerrin Department of Chemical Engineering, Texas A M University, College Station, Texas, United States of America, 2 Department of Biology, Texas A M University, College Station, Texas, United States of America, 3 Zachry Department of Civil Engineering, Texas A M University, College Station, Texas, United States of America Abstract With the opportunistic pathogen Pseudomonas aeruginosa, quorum sensing based on homoserine lactones was found to influence biofilm formation. Here we discern a mechanism by which quorum sensing controls biofilm formation by screening 5850 transposon mutants of P. aeruginosa PA14 for altered biofilm formation. This screen identified the PA3885 mutant, which had 147-fold more biofilm than the wild-type strain. Loss of PA3885 decreased swimming, abolished swarming, and increased attachment, although this did not affect production of rhamnolipids. The PA3885 mutant also had a wrinkly colony phenotype, formed pronounced pellicles, had substantially more aggregation, and had 28-fold more exopolysaccharide production. Expression of PA3885 in trans reduced biofilm formation and abolished aggregation. Whole transcriptome analysis showed that loss of PA3885 activated expression of the pel locus, an operon that encodes for the synthesis of extracellular matrix polysaccharide. Genetic screening identified that loss of PelABDEG and the PA1120 protein (which contains a GGDEF-motif) suppressed the phenotypes of the PA3885 mutant, suggesting that the function of the PA3885 protein is to regulate 3,5-cyclic diguanylic acid