Detailed analysis of the DNA recognition motifs of the Xanthomonas type III effectors AvrBs3 and AvrBs3Deltarep16.
The Gram-negative phytopathogenic bacterium Xanthomonas campestris pv. vesicatoria (Xcv) employs a type III secretion system to translocate effector proteins into plant cells where they modulate host signaling pathways to the pathogen's benefit. The effector protein AvrBs3 acts as a eukaryotic transcription factor and induces the expression of plant genes termed UPA (up-regulated by AvrBs3). Here, we describe 11 new UPA genes from bell pepper that are induced by AvrBs3 early after infection with Xcv. Sequence comparisons revealed the presence of a conserved AvrBs3-responsive element, the UPA box, in all UPA gene promoters analyzed. Analyses of UPA box mutant derivatives confirmed its importance for gene induction by AvrBs3. We show that DNA binding and gene activation were strictly correlated. DNase I footprint studies demonstrated that the UPA box corresponds to the center of the AvrBs3-protected DNA region. Type III delivery of AvrBs3 and mutant derivatives showed that some UPA genes are induced by the AvrBs3 deletion derivative AvrBs3Deltarep16, which lacks four repeats. We show that AvrBs3Deltarep16 recognizes a mutated UPA box with two nucleotide exchanges in positions that are not essential for binding and activation by AvrBs3.
Kay
Sabine
S
Institute of Biology, Department of Genetics, Martin Luther University Halle-Wittenberg, Weinbergweg 10, D-06120 Halle (Saale), Germany.
Hahn
Simone
S
Marois
Eric
E
Wieduwild
Robert
R
Bonas
Ulla
U
D001426:Bacterial Proteins / Q000378:metabolism*; D001483:Base Sequence; D002212:Capsicum / Q000235:genetics* / Q000382:microbiology; D004252:DNA Mutational Analysis; D018076:DNA, Complementary / Q000235:genetics; D018744:DNA, Plant / Q000235:genetics; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D018506:Gene Expression Regulation, Plant; D017343:Genes, Plant; D054884:Host-Pathogen Interactions; D008969:Molecular Sequence Data; D005810:Multigene Family; D010935:Plant Diseases / Q000235:genetics / Q000382:microbiology; D011401:Promoter Regions, Genetic; D011485:Protein Binding; D020218:Response Elements; D000071336:Transcription Activator-Like Effectors; D016959:Xanthomonas campestris / Q000378:metabolism / Q000472:pathogenicity*
D001426:Bacterial Proteins; D018076:DNA, Complementary; D018744:DNA, Plant; D004268:DNA-Binding Proteins; D000071336:Transcription Activator-Like Effectors; C073876:avrBs3-2 protein, Xanthomonas campestris