Novel Role of the Agrobacterium Virulence Effector Protein VirE2 in Modulating Plant Gene Expression
Creators: name: Lapham, Rachelle Amanda
Publisher: name: Purdue University Graduate School
Year: 2019
Subjects: Plant cell and molecular biology
Animal cell and molecular biology
Microbiology not elsewhere classified
Medical biotechnology not elsewhere classified

description: <i>Agrobacterium tumefaciens </i>transfers virulence effector proteins to infected host plants to facilitate the transfer and trafficking of a piece of its tumor inducing (Ti) plasmid, (T-[transfer] DNA), into and through plant cells.<sup> </sup>T-DNA integrates into the host genome where it uses the host’s gene expression machinery to express transgenes. Scientists have used this process to insert beneficial genes into plants by replacing native T-DNA in the bacteria with engineered T-DNA, making <i>Agrobacterium</i>-mediated transformation the preferred method for crop genetic engineering. In spite of its wide-spread use in research and agriculture, we still do not have a complete understanding of the transformation process. Consequently, many important crop species remain highly resistant to transformation. One of my lab’s major goals is to define the molecular interactions between <i>Agrobacterium</i> and its host plants which mediate transformation. I study the role of the <i>Agrobacterium</i> effector protein, VirE2, which is important for plant transformation. VirE2 likely coats the transferred DNA (T-DNA) after it enters the plant cell and protects it from degradation. VIP1 is a host transcription factor that interacts with VirE2 and is involved in activating plant defense responses. VIP1 localizes to both the cytoplasm and the nucleus.<sup> </sup>Under stress, VIP1 localizes to the nucleus where it activates expression of defense response genes.<sup> </sup>This observation led to the model that T-DNA-bound VirE2 binds VIP1 and uses VIP1 nuclear localization to deliver T-DNA into the nucleus (the “Trojan Horse” model). In contrast to this model, our lab has obtained data showing that VirE2 holds at least a portion of the VIP1 pool outside the nucleus. We also showed that VIP1 and its homologs are not necessary for transformation. VirE2 interacts with several host proteins in addition to VIP1, and these interactions could lead to changes in host gene expression and protein levels, possibly facilitating transformation. We investigated this model by placing VirE2 under the control of an inducible promoter in <i>Arabidopsis</i> and performing RNA-seq and proteomics under non-induced and induced conditions, and in the presence of <i>Agrobacterium</i> to determine its individual effect on plant RNA and protein levels during infection. Some genes differentially expressed after VirE2 induction are known to be important for transformation. Knockout mutant lines of some VirE2 differentially expressed genes showed altered transformation phenotypes. Protein levels of genes known to be important for transformation were also increased in response to VirE2 induction, and overexpression of some of these genes resulted in increased transformation susceptibility. We therefore conclude that VirE2 modulates both plant RNA and protein levels to facilitate transformation.