title: Cell-free Escherichia coli-based system to screen for quorum-sensing molecules interacting with quorum receptor proteins of Streptomyces coelicolor.
abstract: Quorum sensing (QS) is mediated by small molecules and involved in diverse cellular functions, such as virulence, biofilm formation, secondary metabolism, and cell differentiation. In this study, we developed a rapid and effective screening tool based on a cell-free Escherichia coli-based expression system to identify QS molecules of Streptomyces. The binding of QS molecules to gamma-butyrolactone receptor ScbR was monitored by changes in the expression levels of the green fluorescent protein reporter in E. coli cell extract. Using this assay system, we could successfully confirm SCB1, a gamma-butyrolactone molecule in Streptomyces coelicolor, binding to its known receptor, ScbR. In addition, we have shown that N-hexanoyl-DL-homoserine lactone, one of the QS molecules in many gram-negative bacteria, can regulate ScbR and trigger precocious antibiotic production in S. coelicolor. Our new method can be applied to other strains for which a screening tool for QS molecules has not yet been developed.
authors.affiliation: School of Chemical and Biological Engineering, Seoul National University, Seoul, Republic of Korea. byungkim@snu.ac.kr.
authors.forename: Yung-Hun
authors.initials: YH
authors.lastname: Yang
authors.forename: Tae-Wan
authors.initials: TW
authors.lastname: Kim
authors.forename: Sung-Hee
authors.initials: SH
authors.lastname: Park
authors.forename: Kwangwon
authors.initials: K
authors.lastname: Lee
authors.forename: Hyung-Yeon
authors.initials: HY
authors.lastname: Park
authors.forename: Eunjung
authors.initials: E
authors.lastname: Song
authors.forename: Hwang-Soo
authors.initials: HS
authors.lastname: Joo
authors.forename: Yun-Gon
authors.initials: YG
authors.lastname: Kim
authors.forename: Ji-Sook
authors.initials: JS
authors.lastname: Hahn
authors.forename: Byung-Gee
authors.initials: BG
authors.lastname: Kim
mesh_terms: D015107:4-Butyrolactone / Q000031:analogs & derivatives* / Q000378:metabolism*; D001426:Bacterial Proteins / Q000378:metabolism*; D015374:Biosensing Techniques; D004268:DNA-Binding Proteins / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics*; D049452:Green Fluorescent Proteins / Q000378:metabolism; D011485:Protein Binding; D053038:Quorum Sensing; D048372:Streptomyces coelicolor / Q000378:metabolism / Q000502:physiology*
chemical_list: D001426:Bacterial Proteins; D004268:DNA-Binding Proteins; C446304:ScbR protein, Streptomyces coelicolor; C088386:homoserine lactone; D049452:Green Fluorescent Proteins; D015107:4-Butyrolactone
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