Novel Xanthomonas campestris long-chain-specific 3-oxoacyl- acyl carrier protein reductase involved in diffusible signal factor synthesis

Zhe Hu, Huijuan Dong, Jin Cheng Ma, Yonghong Yu, Kai Hui Li, Qiao Qiao Guo, Chao Zhang, Wen Bin Zhang, Xinyun Cao, John E. Cronan, Haihong Wang

Research output: Contribution to journalArticle

Abstract

The precursors of the diffusible signal factor (DSF) family signals of Xanthomonas campestris pv. campestris are 3-hydroxyacyl-acyl carrier protein (3- hydroxyacyl-ACP) thioesters having acyl chains of 12 to 13 carbon atoms produced by the fatty acid biosynthetic pathway. We report a novel 3-oxoacyl-ACP reductase encoded by the X. campestris pv. campestris XCC0416 gene (fabG2), which is unable to participate in the initial steps of fatty acyl synthesis. This was shown by the failure of FabG2 expression to allow growth at the nonpermissive temperature of an Escherichia coli fabG temperature-sensitive strain. However, when transformed into the E. coli strain together with a plasmid bearing the Vibrio harveyi acyl-ACP synthetase gene (aasS), growth proceeded, but only when the medium contained octanoic acid. In vitro assays showed that FabG2 catalyzes the reduction of long-chain (≥C8) 3-oxoacyl-ACPs to 3-hydroxyacyl-ACPs but is only weakly active with shorter-chain (C4, C6) substrates. FabG1, the housekeeping 3-oxoacyl-ACP reductase encoded within the fatty acid synthesis gene cluster, could be deleted in a strain that overexpressed fabG2 but only in octanoic acid-supplemented media. Growth of the X. campestris pv. campestris ΔfabG1 strain overexpressing fabG2 required fabH for growth with octanoic acid, indicating that octanoyl coenzyme A is elongated by X. campestris pv. campestris fabH. Deletion of fabG2 reduced DSF family signal production, whereas overproduction of either FabG1 or FabG2 in the ΔfabG2 strain restored DSF family signal levels. IMPORTANCE Quorum sensing mediated by DSF signaling molecules regulates pathogenesis in several different phytopathogenic bacteria, including Xanthomonas campestris pv. campestris. DSF signaling also plays a key role in infection by the human pathogen Burkholderia cepacia. The acyl chains of the DSF molecules are diverted and remodeled from a key intermediate of the fatty acid synthesis pathway. We report a Xanthomonas campestris pv. campestris fatty acid synthesis enzyme, FabG2, of novel specificity that seems tailored to provide DSF signaling molecule precursors.

Original languageEnglish (US)
Article numbere00596-18
JournalmBio
Volume9
Issue number3
DOIs
StatePublished - May 1 2018

Keywords

  • Fatty acids
  • Quorum sensing
  • Xanthomonas

ASJC Scopus subject areas

  • Microbiology
  • Virology

Fingerprint Dive into the research topics of 'Novel Xanthomonas campestris long-chain-specific 3-oxoacyl- acyl carrier protein reductase involved in diffusible signal factor synthesis'. Together they form a unique fingerprint.

  • Cite this

    Hu, Z., Dong, H., Ma, J. C., Yu, Y., Li, K. H., Guo, Q. Q., Zhang, C., Zhang, W. B., Cao, X., Cronan, J. E., & Wang, H. (2018). Novel Xanthomonas campestris long-chain-specific 3-oxoacyl- acyl carrier protein reductase involved in diffusible signal factor synthesis. mBio, 9(3), [e00596-18]. https://doi.org/10.1128/mBio.00596-18