TY - JOUR
T1 - ManA is regulated by RssAB signaling and promotes motility in Serratia marcescens
AU - Soo, Po Chi
AU - Horng, Yu Tze
AU - Chang, Yung Lin
AU - Tsai, Wei Wen
AU - Jeng, Wen Yih
AU - Lu, Chia Chen
AU - Lai, Hsin Chih
N1 - Funding Information:
We appreciate the grants from the National Science Council ( NSC-101-2320-B-320-002 ), CMRPG361231 and CMRPD170101 from Chang Gung Memorial Hospital, Taiwan , which supported this work. We thank Mr. I-Jen Hsieh for TLC technique support.
PY - 2014/1
Y1 - 2014/1
N2 - Serratia marcescens swarms on 0.8% LB agar at 30°C but not at 37°C. To understand the molecular mechanism regulating Serratia swarming, transposon mutagenesis was performed to screen for mutants that swarmed at 37°C. In one mutant, S. marcescens WW100, the transposon was inserted in the upstream region of manA, which encodes mannose-6-phosphate isomerase, a type I phosphomannose isomerase. The transcriptional and translational levels of manA were higher in S. marcescens WW100 than in the wild-type strain. S. marcescens WW100 produced more serrawettin W1 (biosurfactant) than the wild-type, as detected by thin-layer chromatography, to promote surface motility by reducing surface tension. Serratia swarming was previously shown to be negatively regulated by the RssA-RssB two-component system. An electrophoretic mobility shift assay (EMSA) indicated that phosphorylated RssB (the response regulator) binds upstream of the transposon insertion site and manA in S. marcescens WW100. Analysis by real-time RT-PCR (qRT-PCR) revealed that, compared to the wild-type level, manA mRNA was increased in the rssA deletion mutant. The results indicated that RssA-RssB signaling directly represses the expression of manA and that overexpression of manA increases the production of serrawettin for Serratia swarming at 37°C.
AB - Serratia marcescens swarms on 0.8% LB agar at 30°C but not at 37°C. To understand the molecular mechanism regulating Serratia swarming, transposon mutagenesis was performed to screen for mutants that swarmed at 37°C. In one mutant, S. marcescens WW100, the transposon was inserted in the upstream region of manA, which encodes mannose-6-phosphate isomerase, a type I phosphomannose isomerase. The transcriptional and translational levels of manA were higher in S. marcescens WW100 than in the wild-type strain. S. marcescens WW100 produced more serrawettin W1 (biosurfactant) than the wild-type, as detected by thin-layer chromatography, to promote surface motility by reducing surface tension. Serratia swarming was previously shown to be negatively regulated by the RssA-RssB two-component system. An electrophoretic mobility shift assay (EMSA) indicated that phosphorylated RssB (the response regulator) binds upstream of the transposon insertion site and manA in S. marcescens WW100. Analysis by real-time RT-PCR (qRT-PCR) revealed that, compared to the wild-type level, manA mRNA was increased in the rssA deletion mutant. The results indicated that RssA-RssB signaling directly represses the expression of manA and that overexpression of manA increases the production of serrawettin for Serratia swarming at 37°C.
UR - http://www.scopus.com/inward/record.url?scp=84893812383&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84893812383&partnerID=8YFLogxK
U2 - 10.1016/j.resmic.2013.10.005
DO - 10.1016/j.resmic.2013.10.005
M3 - Article
C2 - 24161484
AN - SCOPUS:84893812383
VL - 165
SP - 21
EP - 29
JO - Research in Microbiology
JF - Research in Microbiology
SN - 0923-2508
IS - 1
ER -