Research Accomplishment Reports 2007

Genomics, Molecular Biology and Cell Biology of Sonchus yellow net virus, a Plant Rhabdovirus

M.M. Goodin
Department of Plant Pathology

 

Project Description

Elucidation of the mechanism by which viruses establish a compatible interaction with their hosts, in addition to being the long-term research goal of the Goodin lab, is a fundamental requirement for the development of novel strategies to prevent virus infection and for engineering improved varieties of host plants to be used in pharmaceutical applications that employ viral vectors.

Our immediate research focuses on the cell biology of plant-infecting members of the rhabdoviridae, which are enveloped viruses with monopartite, minus-sense, single-stranded MA genomes that include some of the greatest threats to human, animal, and plant health. In experiments utilizing sonchus yellow net virus (SYNV) and potato yellow dwarf virus (PYDV) as paradigms for cell biology studies of rhabdoviruses that replicate in nuclei of infected cells, we initially characterized virus-induced changes in plant nuclear membranes. In order to advance these studies we have constructed a novel set of binary vectors for the expression of autofluorescent protein fusions in plants. Using these vectors in conjunction with advanced confocal imaging techniques, we have produced the most detailed map of protein localization in rhabdovirus-infected cells constructed to date. 

To advance these studies, we are developing many new vectors for the expression of fluorescent protein fusions in plant cells. Additionally, we are engineering several lines of transgenic plants that express fluorescent markers for a variety of cellular loci. We plan to integrate our cell biology studies with microarray experiments that have identified over 1,400 genes for which expression changes in response to infection by SYNV.

Finally, we have recently determined the nucleotide sequences of the genes encoding the nucleocapsid (n) and phosphoprotein (p) proteins of PYDV. Full-length CDNA clones for these genes will be mobilized into our novel expression vectors. Expression of autofluorescent fusions of the pydv-n and -p proteins will be used to characterize their cellular localization and interaction. Given the differential effects of SYNV and PYDV on nuclear membranes of host cells, we anticipate that these, and future, protein localization studies will provide novel insights into the cell biology of these viruses.

Impact

The broader impact of these studies will be to establish a paradigm for plant/rhabdovirus interactions, in addition to deciphering the biological principles that govern such plant/virus interactions. The information acquired in these studies will lead to the development of novel methods for controlling viral diseases of plants. Additionally, this information can be used to develop new, genetically-engineered plants that are optimized for their ability to produce pharmaceutical proteins. Such plants would be ideal hosts for recombinant viruses that express proteins in plants that can be used in the fight against such diseases as cancer and arthritis, as well as, complications that stem from tissue rejection following organ transplant.

Publications

Goodin, M.M., Chakrabarty, R., Banerjee, R., Yelton, S., DeBolt, S. (2007) Update on live-cell imaging in plant cells: New Gateways to discovery. Plant Physiology, 145:1100-1109.

Chakrabarty, C., Banerjee, R., Chung, S-M., Farman, M., Citovsky, V., Hogenhout, S.A., Tzfira, T., and Goodin, M.M. (2007)  pSITE Vectors for Stable Integration or Transient Expression of Autofluorescent Protein Fusions in Plants: Probing Nicotiana benthamiana-Virus Interactions. MPMI. 20:740-750

Goodin, M.M., Chakrabarty, R., Yelton, S., Martin, K., Clark, A., Brooks, R. (2007) Membrane and protein dynamics in live plant nuclei infected with Sonchus yellow net virus, a plant-adapted rhabdovirus. J Gen Virol. 88:1810-1820.

Deng, M., Bragg, J.N., Ruzin, S., Schichnes, D., King, D., Goodin, M.M., Jackson, A.O. (2007) Role of the sonchus yellow net virus N protein in formation of nuclear viroplasms. J Virol. 81:5362-5374.