The Ralstonia solanacearum pathogenicity regulator HrpB induces 3-hydroxy-oxindole synthesis.
The transcriptional activator HrpB of the bacterial wilt causing betaproteobacterium Ralstonia solanacearum represents a key regulator for pathogenicity. In particular, it drives expression of hrp genes encoding a type III secretion system (T3SS) as well as effector molecules for delivery into the host cytosol to promote disease. However, the HrpB regulon extends beyond this T3SS. We describe here an HrpB-activated operon of six genes that is responsible for the synthesis of a fluorescent isatin derivative of 149 Amu that we named HDF for HrpB-dependent factor and that we purified from culture supernatants. The structure of the labile molecule was solved by using NMR and CD spectroscopy to be (3S)-3-hydroxy-indolin-2-one and confirmed by its chemical synthesis and MS spectrometry. HDF was found to be present at 20 nM in wild-type cultures grown on minimal medium, and its synthesis increased 15-fold upon overproduction of HrpB, confirming that HrpB activates HDF synthesis. The addition of tryptophan significantly stimulated HDF biosynthesis and was shown to represent the precursor molecule for HDF synthesis. A search for the biological function of the molecule revealed that HDF induces acyl-homoserine lactone receptor-mediated reporter activity of the well studied LuxR transcriptional regulator of Vibrio fischeri. Thus, our results provide evidence that the specificity of acyl-homoserine lactone (acyl-HSL) receptors is clearly broader than previously considered. The failure to detect induction by HDF of the described endogenous quorum-sensing circuits of the pathogen points to a role in interfering with cell-cell signaling of rivalling bacteria.
Delaspre
Fabien
F
Laboratoire des Interactions Plantes Micro-Organismes (LIPM), Institut National de la Recherche Agronomique/Centre National de la Recherche Scientifique, BP52627, 31326 Castanet-Tolosan, France.
Nieto Peñalver
Carlos G
CG
Saurel
Olivier
O
Kiefer
Patrick
P
Gras
Emmanuel
E
Milon
Alain
A
Boucher
Christian
C
Genin
Stéphane
S
Vorholt
Julia A
JA
D001426:Bacterial Proteins / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*; D007211:Indoles / Q000378:metabolism*; D009682:Magnetic Resonance Spectroscopy; D009154:Mutation; D009876:Operon; D010935:Plant Diseases / Q000382:microbiology; D043368:Ralstonia solanacearum / Q000235:genetics / Q000472:pathogenicity*; D014157:Transcription Factors / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*; D014364:Tryptophan / Q000378:metabolism
D001426:Bacterial Proteins; D004268:DNA-Binding Proteins; D007211:Indoles; D014157:Transcription Factors; C079047:hrpB protein, Ralstonia solanacearum; D014364:Tryptophan; C510549:3-hydroxyindole