Agrobacterium VirE2 Protein Modulates Plant Gene Expression and Mediates Transformation from Its Location in the Cytoplasm

Agrobacterium effector protein VirE2 is important for plant transformation. VirE2 likely coats transferred DNA (T-DNA) in the plant cell and protects it from degradation. VirE2 localizes to the plant cytoplasm and interacts with several host proteins. Plant-expressed VirE2 can complement a virE2 mutant Agrobacterium strain to support transformation. We investigated whether VirE2 could facilitate transformation from a nuclear location by affixing to it a strong nuclear localization signal (NLS) sequence. Only cytoplasmic-, but not nuclear-localized, VirE2 could stimulate transformation. To investigate the ways VirE2 supports transformation, we generated transgenic Arabidopsis plants containing a virE2 gene under the control of an inducible promoter and performed RNA-seq and proteomic analyses before and after induction. Some differentially expressed plant genes were previously known to facilitate transformation. Knockout mutant lines of some other VirE2 differentially expressed genes showed altered transformation phenotypes. Levels of some proteins known to be important for transformation increased in response to VirE2 induction, but prior to or without induction of their corresponding mRNAs. Overexpression of some other genes whose proteins increased after VirE2 induction resulted in increased transformation susceptibility. We conclude that cytoplasmically-localized VirE2 modulates both plant RNA and protein levels to facilitate transformation. Overall design: Examination of differentially expressed genes using RNAseq in transgenic Arabidopsis thaliana roots which inducibly express the Agrobacterium tumefaciens virulence protein VirE2 under induced and non-induced conditions in the presence of an avirulent Agrobacterium strain at 0, 3, and 12 hours post-treatment