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The Type III Effector HsvG of the Gall-Forming Pantoea agglomerans Mediates Expression of the Host Gene HSVGT

February 2012 , Volume 25 , Number  2
Pages  231 - 240

Gal Nissan,1 Shulamit Manulis-Sasson,2 Laura Chalupowicz,2 Doron Teper,1 Adva Yeheskel,3 Metsada Pasmanik-Chor,3 Guido Sessa,1 and Isaac Barash1

1Department of Molecular Biology and Ecology of Plants, Faculty of Life Sciences, Tel-Aviv University, Tel-Aviv 69978, Israel; 2Department of Plant Pathology and Weed Research, ARO, The Volcani Center, Bet Dagan 50250, Israel; 3Bioinformatics Unit, Faculty of Life Sciences, Tel-Aviv University, Tel-Aviv 69978, Israel

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Accepted 4 October 2011.

The type III effector HsvG of the gall-forming Pantoea agglomerans pv. gypsophilae is a DNA-binding protein that is imported to the host nucleus and involved in host specificity. The DNA-binding region of HsvG was delineated to 266 amino acids located within a secondary structure region near the N-terminus of the protein but did not display any homology to canonical DNA-binding motifs. A binding site selection procedure was used to isolate a target gene of HsvG, named HSVGT, in Gypsophila paniculata. HSVGT is a predicted acidic protein of the DnaJ family with 244 amino acids. It harbors characteristic conserved motifs of a eukaryotic transcription factor, including a bipartite nuclear localization signal, zinc finger, and leucine zipper DNA-binding motifs. Quantitative real-time polymerase chain reaction analysis demonstrated that HSVGT transcription is specifically induced in planta within 2 h after inoculation with the wild-type P. agglomerans pv. gypsophilae compared with the hsvG mutant. Induction of HSVGT reached a peak of sixfold at 4 h after inoculation and progressively declined thereafter. Gel-shift assay demonstrated that HsvG binds to the HSVGT promoter, indicating that HSVGT is a direct target of HsvG. Our results support the hypothesis that HsvG functions as a transcription factor in gypsophila.

© 2012 The American Phytopathological Society