title: Homologous expression and T3SS-dependent secretion of TAP-tagged Xo2276 in Xanthomonas oryzae pv. oryzae induced by rice leaf extract and its direct in vitro recognition of putative target DNA sequence.
abstract: Xo2276 is a putative transcription activator-like effector (TALE) in Xanthomonas oryzae pv. oryzae (Xoo). Xo2276 was expressed with a TAP-tag at the C-terminus in Xoo cells to enable quantitative analysis of protein expression and secretion. Nearly all TAP-tagged Xo2276 existed in an insoluble form; addition of rice leaf extracts from a Xoo-susceptible rice cultivar, Milyang23, significantly stimulated secretion of TAP-tagged Xo2276 into the medium. In a T3SS-defective Xoo mutant strain, secretion of TAP-tagged Xo2276 was blocked. Xo2276 is a Xoo ortholog of Xanthomonas campestris pv. vesicatoria (Xcv) AvrBs3 and contains a conserved DNA-binding domain (DBD), which includes 19.5 tandem repeats of 34 amino acids. Xo2276- DBD was expressed in E. coli and purified. Direct in vitro recognition of Xo2276-DBD on a putative target DNA sequence was confirmed using an electrophoretic mobility shift assay. This is the first study measuring the homologous expression and secretion of Xo2276 in vitro using rice leaf extract and its direct in vitro binding to the specific target DNA sequence.
authors: Department of Advanced Technology Fusion, Konkuk University, Seoul 143-701, Korea.
authors: Seunghwan
authors: S
authors: Kim
authors: Thi-Dieu-Hanh
authors: TD
authors: Nguyen
authors: Joohee
authors: J
authors: Lee
authors: Myoung-ki
authors: MK
authors: Hong
authors: Tan-Viet
authors: TV
authors: Pham
authors: Yeh-Jin
authors: YJ
authors: Ahn
authors: Byoung-Moo
authors: BM
authors: Lee
authors: Ye Sun
authors: YS
authors: Han
authors: Dong-Eun
authors: DE
authors: Kim
authors: Jeong-Gu
authors: JG
authors: Kim
authors: Lin-Woo
authors: LW
authors: Kang
mesh_terms: D004269:DNA, Bacterial / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000235:genetics* / Q000378:metabolism*; D024202:Electrophoretic Mobility Shift Assay; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D020869:Gene Expression Profiling; D012275:Oryza / Q000737:chemistry; D010936:Plant Extracts / Q000302:isolation & purification / Q000378:metabolism; D018515:Plant Leaves / Q000737:chemistry; D011485:Protein Binding; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D014974:Xanthomonas / Q000235:genetics* / Q000378:metabolism*
chemical_list: D004269:DNA, Bacterial; D004268:DNA-Binding Proteins; D010936:Plant Extracts; D011994:Recombinant Proteins