Genomewide identification of Pseudomonas syringae pv. tomato DC3000 promoters controlled by the HrpL alternative sigma factor
Journal
Proceedings of the National Academy of Sciences of the United States of America
Journal Volume
99
Journal Volume
99
Journal Issue
4
Journal Issue
4
Pages
2275-2280
Start Page
2275
End Page
2280
ISSN
00278424
Date Issued
2002-02-19
Author(s)
Fouts, Derrick E.
Abramovitch, Robert B.
Alfano, James R.
Baldo, Angela M.
Buell, C. Robin
Cartinhour, Samuel
Chatterjee, Arun K.
D'Ascenzo, Mark
Gwinn, Michelle L.
Lazarowitz, Sondra G.
Martin, Gregory B.
Rehm, Amos H.
Schneider, David J.
Van Dijk, Karin
Tang, Xiaoyan
Collmer, Alan
Abstract
The ability of Pseudomonas syringae pv. tomato DC3000 to parasitize tomato and Arabidopsis thaliana depends on genes activated by the HrpL alternative sigma factor. To support various functional genomic analyses of DC3000, and specifically, to identify genes involved in pathogenesis, we developed a draft sequence of DC3000 and used an iterative process involving computational and gene expression techniques to identify virulence-implicated genes downstream of HrpL-responsive promoters. Hypersensitive response and pathogenicity (Hrp) promoters are known to control genes encoding the Hrp (type III protein secretion) machinery and a few type III effector proteins in DC3000. This process involved (i) identification of 9 new virulence-implicated genes in the Hrp regulon by miniTn5gus mutagenesis, (ii) development of a hidden Markov model (HMM) trained with known and transposon-identified Hrp promoter sequences, (iii) HMM identification of promoters upstream of 12 additional virulence-implicated genes, and (iv) microarray and RNA blot analyses of the HrpL-dependent expression of a representative subset of these DC3000 genes. We found that the Hrp regulon encodes candidates for 4 additional type III secretion machinery accessory factors, homologs of the effector proteins HopPsyA, AvrPpiB1 (2 copies), AvrPpiC2, AvrPphD (2 copies), AvrPphE, AvrPphF, and AvrXv3, and genes associated with the production or metabolism of virulence factors unrelated to the Hrp type III secretion system, including syringomycin synthetase (SyrE), N(epsilon)-(indole-3-acetyl)-l-lysine synthetase (IaaL), and a subsidiary regulon controlling coronatine production. Additional candidate effector genes, hopPtoA2, hopPtoB2, and an avrRps4 homolog, were preceded by Hrp promoter-like sequences, but these had HMM expectation values of relatively low significance and were not detectably activated by HrpL.
Publisher
National Academy of Sciences
Type
journal article