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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

The effects of isoleucine on IDSF biosynthesis.(a) HPLC analysis of DSF, BDSF and IDSF production in media XYS and XYSI. XYSI: XYS medium supplemented with 50 μM isoleucine. (b) The dose–response curve between isoleucine concentrations (10 to 75 μM) and IDSF production. (c) IDSF production time course of the ΔrpfC strain in XYSI medium. Data are means ± one standard deviation of three independent assays.
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f4: The effects of isoleucine on IDSF biosynthesis.(a) HPLC analysis of DSF, BDSF and IDSF production in media XYS and XYSI. XYSI: XYS medium supplemented with 50 μM isoleucine. (b) The dose–response curve between isoleucine concentrations (10 to 75 μM) and IDSF production. (c) IDSF production time course of the ΔrpfC strain in XYSI medium. Data are means ± one standard deviation of three independent assays.

Mentions: Supplementation of medium XSY with isoleucine (50 μM) led to IDSF production and a slight decrease in DSF and BDSF production (Fig. 4a). IDSF levels increased proportionately with increasing concentrations (10–75 μM) of isoleucine (Fig. 4b). The ΔrpfC strain produced 0.42 μM (1.4 × 10−3 μmol/109 cells) of IDSF in medium XYS medium supplemented with 50 μM isoleucine (XYSI) after 24 h of growth (Fig. 4c).


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)

The effects of isoleucine on IDSF biosynthesis.(a) HPLC analysis of DSF, BDSF and IDSF production in media XYS and XYSI. XYSI: XYS medium supplemented with 50 μM isoleucine. (b) The dose–response curve between isoleucine concentrations (10 to 75 μM) and IDSF production. (c) IDSF production time course of the ΔrpfC strain in XYSI medium. Data are means ± one standard deviation of three independent assays.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

f4: The effects of isoleucine on IDSF biosynthesis.(a) HPLC analysis of DSF, BDSF and IDSF production in media XYS and XYSI. XYSI: XYS medium supplemented with 50 μM isoleucine. (b) The dose–response curve between isoleucine concentrations (10 to 75 μM) and IDSF production. (c) IDSF production time course of the ΔrpfC strain in XYSI medium. Data are means ± one standard deviation of three independent assays.
Mentions: Supplementation of medium XSY with isoleucine (50 μM) led to IDSF production and a slight decrease in DSF and BDSF production (Fig. 4a). IDSF levels increased proportionately with increasing concentrations (10–75 μM) of isoleucine (Fig. 4b). The ΔrpfC strain produced 0.42 μM (1.4 × 10−3 μmol/109 cells) of IDSF in medium XYS medium supplemented with 50 μM isoleucine (XYSI) after 24 h of growth (Fig. 4c).

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