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The Multiple DSF-family QS Signals are Synthesized from Carbohydrate and Branched-chain Amino Acids via the FAS Elongation Cycle.

Zhou L, Yu Y, Chen X, Diab AA, Ruan L, He J, Wang H, He YW - Sci Rep (2015)

Bottom Line: Furthermore, our biochemical analyses show that the key DSF synthase RpfF has both thioesterase and dehydratase activities, and uses 3-hydroxydedecanoyl-ACP as a substrate to produce BDSF.Finally, our results show that the classic fatty acid synthesis elongation cycle is required for the biosynthesis of DSF-family signals.Taken all together, these findings establish a general biosynthetic pathway for the DSF-family quorum sensing signals.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Microbial Metabolism, School of Life Sciences &Biotechnology, Shanghai Jiao Tong University, Shanghai 200240, China.

ABSTRACT
Members of the diffusible signal factor (DSF) family are a novel class of quorum sensing (QS) signals in diverse Gram-negative bacteria. Although previous studies have identified RpfF as a key enzyme for the biosynthesis of DSF family signals, many questions in their biosynthesis remain to be addressed. In this study with the phytopathogen Xanthomonas campestris pv. campestris (Xcc), we show that Xcc produces four DSF-family signals (DSF, BDSF, CDSF and IDSF) during cell culture, and that IDSF is a new functional signal characterized as cis-10-methyl-2-dodecenoic acid. Using a range of defined media, we further demonstrate that Xcc mainly produces BDSF in the presence of carbohydrates; leucine and valine are the primary precursor for DSF biosynthesis; isoleucine is the primary precursor for IDSF biosynthesis. Furthermore, our biochemical analyses show that the key DSF synthase RpfF has both thioesterase and dehydratase activities, and uses 3-hydroxydedecanoyl-ACP as a substrate to produce BDSF. Finally, our results show that the classic fatty acid synthesis elongation cycle is required for the biosynthesis of DSF-family signals. Taken all together, these findings establish a general biosynthetic pathway for the DSF-family quorum sensing signals.

No MeSH data available.


Related in: MedlinePlus

DSF-family signal production of Xcc.(a) HPLC analysis of the compounds in fractions A–D extracted from the strain ΔrpfC culture supernatant. (b) The chemical structures of DSF, BDSF, CDSF and IDSF as determined by liquid chromatography-mass spectrometry and/or nuclear magnetic resonance analysis. (c) The proportions of DSF, BDSF, CDSF and IDSF produced by strain ΔrpfC supernatant in media YEB, NA, NYG, and XOLN.
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f1: DSF-family signal production of Xcc.(a) HPLC analysis of the compounds in fractions A–D extracted from the strain ΔrpfC culture supernatant. (b) The chemical structures of DSF, BDSF, CDSF and IDSF as determined by liquid chromatography-mass spectrometry and/or nuclear magnetic resonance analysis. (c) The proportions of DSF, BDSF, CDSF and IDSF produced by strain ΔrpfC supernatant in media YEB, NA, NYG, and XOLN.

Mentions: DSF-family signals produced by Xcc strain XC1 (rpfC) in NA medium were extracted as previously described6. Further separation with HPLC identified four active fractions with retention times of 14.3 (C), 15.2 (B), 20.3 (D), and 21.1 (A) min respectively, which showed UV absorption maxima at 212 nm and strong DSF activity (Fig. 1a and Supplementary Fig. S1c). Liquid chromatography-mass spectrometry (LC-MS) analysis confirmed that the three compounds in peaks A, B and C were the previously identified DSF-family signals DSF, BDSF and CDSF (Fig. 1b and Supplementary Fig. S1d–f). The spectrometry data of fraction D suggested a new member of DSF-family signals (IDSF), the characterization of which will be discussed in the following section.


The Multiple DSF-family QS Signals are Synthesized from Carbohydrate and Branched-chain Amino Acids via the FAS Elongation Cycle.

Zhou L, Yu Y, Chen X, Diab AA, Ruan L, He J, Wang H, He YW - Sci Rep (2015)

DSF-family signal production of Xcc.(a) HPLC analysis of the compounds in fractions A–D extracted from the strain ΔrpfC culture supernatant. (b) The chemical structures of DSF, BDSF, CDSF and IDSF as determined by liquid chromatography-mass spectrometry and/or nuclear magnetic resonance analysis. (c) The proportions of DSF, BDSF, CDSF and IDSF produced by strain ΔrpfC supernatant in media YEB, NA, NYG, and XOLN.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4542539&req=5

f1: DSF-family signal production of Xcc.(a) HPLC analysis of the compounds in fractions A–D extracted from the strain ΔrpfC culture supernatant. (b) The chemical structures of DSF, BDSF, CDSF and IDSF as determined by liquid chromatography-mass spectrometry and/or nuclear magnetic resonance analysis. (c) The proportions of DSF, BDSF, CDSF and IDSF produced by strain ΔrpfC supernatant in media YEB, NA, NYG, and XOLN.
Mentions: DSF-family signals produced by Xcc strain XC1 (rpfC) in NA medium were extracted as previously described6. Further separation with HPLC identified four active fractions with retention times of 14.3 (C), 15.2 (B), 20.3 (D), and 21.1 (A) min respectively, which showed UV absorption maxima at 212 nm and strong DSF activity (Fig. 1a and Supplementary Fig. S1c). Liquid chromatography-mass spectrometry (LC-MS) analysis confirmed that the three compounds in peaks A, B and C were the previously identified DSF-family signals DSF, BDSF and CDSF (Fig. 1b and Supplementary Fig. S1d–f). The spectrometry data of fraction D suggested a new member of DSF-family signals (IDSF), the characterization of which will be discussed in the following section.

Bottom Line: Furthermore, our biochemical analyses show that the key DSF synthase RpfF has both thioesterase and dehydratase activities, and uses 3-hydroxydedecanoyl-ACP as a substrate to produce BDSF.Finally, our results show that the classic fatty acid synthesis elongation cycle is required for the biosynthesis of DSF-family signals.Taken all together, these findings establish a general biosynthetic pathway for the DSF-family quorum sensing signals.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Microbial Metabolism, School of Life Sciences &Biotechnology, Shanghai Jiao Tong University, Shanghai 200240, China.

ABSTRACT
Members of the diffusible signal factor (DSF) family are a novel class of quorum sensing (QS) signals in diverse Gram-negative bacteria. Although previous studies have identified RpfF as a key enzyme for the biosynthesis of DSF family signals, many questions in their biosynthesis remain to be addressed. In this study with the phytopathogen Xanthomonas campestris pv. campestris (Xcc), we show that Xcc produces four DSF-family signals (DSF, BDSF, CDSF and IDSF) during cell culture, and that IDSF is a new functional signal characterized as cis-10-methyl-2-dodecenoic acid. Using a range of defined media, we further demonstrate that Xcc mainly produces BDSF in the presence of carbohydrates; leucine and valine are the primary precursor for DSF biosynthesis; isoleucine is the primary precursor for IDSF biosynthesis. Furthermore, our biochemical analyses show that the key DSF synthase RpfF has both thioesterase and dehydratase activities, and uses 3-hydroxydedecanoyl-ACP as a substrate to produce BDSF. Finally, our results show that the classic fatty acid synthesis elongation cycle is required for the biosynthesis of DSF-family signals. Taken all together, these findings establish a general biosynthetic pathway for the DSF-family quorum sensing signals.

No MeSH data available.


Related in: MedlinePlus