Coreplication experiments have revealed that replication of
turnip yellow mosaic virus (TYMV) RNA in turnip protoplasts is cis-preferential.
Genomes encoding mutant p141 or p66, proteins
essential for virus replication, were inefficiently rescued by a
helper genome. One model for the cis-preferential replication of
TYMV is that p66 and p141 form a complex that associates with the
RNA from which they are translated, limiting their availability in
trans. Three types of experiments were used in this study in an
attempt to obtain physical evidence for the hypothetical interaction
between p66 and p141. Immunoprecipitations from in vitro
translation reactions using antiserum that recognizes p66 (and its
progenitor, p206) coprecipitate p141, indicating that the proteins
form a complex in vitro. The results of coimmunoprecipitations of
translation products with in-frame deletions did not lead to
definitive information about interaction domains. p66 and the
helicase domain of p141 do not detectably interact in the yeast two-hybrid
system or in GST fusion interaction assays. Problems with
the expression of full length p141 fusions make conclusions about
the interaction of other p141 domains with p66 not possible at this
time. Since the helicase domain of p141 does not appear to interact
with p66, future experiments will focus on obtaining expression of
smaller domains of p141, outside the helicase domain, and
determining if they interact with p66. Variations to the model that
do not necessitate the direct interaction between p66 and p141 are
also considered. / Graduation date: 1997
| Identifer | oai:union.ndltd.org:ORGSU/oai:ir.library.oregonstate.edu:1957/34184 |
| Date | 28 January 1997 |
| Creators | Wallace, S. Ellen |
| Contributors | Dreher, Theo W. |
| Source Sets | Oregon State University |
| Language | en_US |
| Detected Language | English |
| Type | Thesis/Dissertation |
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