ManA is regulated by RssAB signaling and promotes motility in Serratia marcescens

Po Chi Soo, Yu Tze Horng, Yung Lin Chang, Wei Wen Tsai, Wen-Yih Jeng, Chia Chen Lu, Hsin Chih Lai

Research output: Contribution to journalArticle

9 Citations (Scopus)

Abstract

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.

Original languageEnglish
Pages (from-to)21-29
Number of pages9
JournalResearch in Microbiology
Volume165
Issue number1
DOIs
Publication statusPublished - 2014 Jan 1

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Serratia marcescens
Serratia
Mannose-6-Phosphate Isomerase
Surface Tension
Electrophoretic Mobility Shift Assay
Thin Layer Chromatography
Mutagenesis
Agar
Real-Time Polymerase Chain Reaction
Polymerase Chain Reaction
Messenger RNA

All Science Journal Classification (ASJC) codes

  • Microbiology
  • Molecular Biology

Cite this

Soo, Po Chi ; Horng, Yu Tze ; Chang, Yung Lin ; Tsai, Wei Wen ; Jeng, Wen-Yih ; Lu, Chia Chen ; Lai, Hsin Chih. / ManA is regulated by RssAB signaling and promotes motility in Serratia marcescens. In: Research in Microbiology. 2014 ; Vol. 165, No. 1. pp. 21-29.
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abstract = "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.",
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ManA is regulated by RssAB signaling and promotes motility in Serratia marcescens. / Soo, Po Chi; Horng, Yu Tze; Chang, Yung Lin; Tsai, Wei Wen; Jeng, Wen-Yih; Lu, Chia Chen; Lai, Hsin Chih.

In: Research in Microbiology, Vol. 165, No. 1, 01.01.2014, p. 21-29.

Research output: Contribution to journalArticle

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

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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.

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