Protein Interactions During Replication of a Plant Rhabodovirus

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; R o o t E n t r y F l; C o m p O b j b W o r d D o c u m e n t O b j e c t P o o l l; l; 8 9 : ; = ? @ A F Microsoft Word 6.0 Document MSWordDoc Word.Document.6 ; 9418086 Jackson This proposal addresses the intracellular location of viral proteins involved in replication and packaging the plant rhabdovirus, sonchus yellow net virus (SYNV), and the biochemistry of the events occurring during replication of the virus in the plant nucleus and subsequent budding from the nuclear membrane. This is a continuing study from one of the few labs in the world studying this important class of enveloped virus in plants at the cellular and molecular level. The biology of SYNV challenges the paradigms of animal rhabdovirus biology. Unlike the animal rhabdovirus, Vesicular Stomatitis Virus (VSV), which replicates in cytoplasm and buds from the cell membrane, thereby serving as a model for secretory membrane traffic, SYNV replicates in the nucleus and buds from the nuclear membrane . This proposal will study these differences, including examining the nature of a gene product, sc4, not found in animal rhabdoviruses. The nuclear targeting of this product, which lacks a transmembrane domain, is of particular interest. The nature of the nuclear targeting domains of th e other proteins is of interest in order to lay a firm groundwork for this system. Since replication occurs in the nucleus, potentially novel aspects of the replication complex of SYNV will be explored through the analysis of protein-protein interaction of transcription-active proteins and the comparison of the transcriptionally inactive core particles from purified virus with transcriptionally active nuclear complexes. %%% Information on plant Rhabdovirusues is limited in comparison to their animal counterparts. Yet, as has been shown in the case of other groups that encompass both animal and plant viruses, such as the picorna- and sindbis-like virus groups, and the Bunyaviridae, the study in parallel members of both kingdoms has often led to a much more rapid development of our understanding of replication of these viruses. Investigations into targeting the components of this virus into the nucleus are also timely because the mechanism of targeting select proteins into the lumen of the nucleus and of integration of proteins into the nuclear membrane of plant and animal cells is unclear. There is the potential here for developing model proteins to examine targeting to the inner nuclear envelope, an area where there is very little knowledge. The plant rhabdoviruses could also provide a valuable system to examine nuclear/cytoplasmic flow of protein and the intracellular flow of virion-containing vesicles in plants. *** ; Oh +' 0 S u m m a r y I n f o r m a t i o n ( 7 $ H l D h R:\WWUSER\TEMPLATE\NORMAL.DOT Lawrence R. Griffing Una Solomon @ g @ @ @ Microsoft Word 6.0 4 e = e p p p p p p p C 1 Õ % t T 3 C p C p p p p Õ p p p p Õ 7 9418086 Jackson This proposal addresses the intracellular lo