title: Direct fluorescence detection of VirE2 secretion by Agrobacterium tumefaciens.
abstract: VirE2 is a ssDNA binding protein essential for virulence in Agrobacterium tumefaciens. A tetracysteine mutant (VirE2-TC) was prepared for in vitro and in vivo fluorescence imaging based on the ReAsH reagent. VirE2-TC was found to be biochemically active as it binds both ssDNA and the acidic secretion chaperone VirE1. It was also biologically functional in complementing virE2 null strains transforming Arabidopsis thaliana roots and Nicotiana tabacum leaves. In vitro experiments demonstrated a two-color fluorescent complex using VirE2-TC/ReAsH and Alexa Fluor 488 labeled ssDNA. In vivo, fluorescent VirE2-TC/ReAsH was detected in bacteria and in plant cells at time frames relevant to transformation.
authors.affiliation: Dept of Materials and Interfaces, Weizmann Institute of Science, Rehovot, Israel.
authors.forename: Noga
authors.initials: N
authors.lastname: Yaakov
authors.affiliation: Chemical Research Support Dept, Weizmann Institute of Science, Rehovot, Israel.
authors.forename: Yoav
authors.initials: Y
authors.lastname: Barak
authors.affiliation: Dept of Plant and Environmental Sciences, Weizmann Institute of Science, Rehovot, Israel.
authors.forename: Idan
authors.initials: I
authors.lastname: Pereman
authors.affiliation: Department of Microbiology and Molecular Genetics, UT-Houston Medical School, Houston, Texas, United States of America.
authors.forename: Peter J
authors.initials: PJ
authors.lastname: Christie
authors.affiliation: Dept of Materials and Interfaces, Weizmann Institute of Science, Rehovot, Israel.
authors.forename: Michael
authors.initials: M
authors.lastname: Elbaum
mesh_terms: D016960:Agrobacterium tumefaciens / Q000166:cytology / Q000235:genetics / Q000378:metabolism*; D017360:Arabidopsis / Q000382:microbiology*; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D004277:DNA, Single-Stranded / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D009154:Mutation; D061848:Optical Imaging; D010941:Plant Tumors / Q000382:microbiology*; D011485:Protein Binding; D014026:Nicotiana / Q000382:microbiology*; D037521:Virulence Factors / Q000235:genetics / Q000378:metabolism*
chemical_list: D001426:Bacterial Proteins; D004277:DNA, Single-Stranded; D004268:DNA-Binding Proteins; D037521:Virulence Factors