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Vol. 11, Issue 8, 2617-2629, August 2000
Department of Biochemistry, Emory University School of Medicine,
Atlanta, GA 30322
The small GTPase Ran is required for the trafficking of
macromolecules into and out of the nucleus. Ran also has been
implicated in cell cycle control, specifically in mitotic spindle
assembly. In interphase cells, Ran is predominately nuclear and thought to be GTP bound, but it is also present in the cytoplasm, probably in
the GDP-bound state. Nuclear transport factor 2 (NTF2) has been shown
to import RanGDP into the nucleus. Here, we examine the in vivo role of
NTF2 in Ran import and the effect that disruption of Ran imported into
the nucleus has on the cell cycle. A temperature-sensitive (ts)
mutant of Saccharomyces cerevisiae NTF2 that does not
bind to Ran is unable to import Ran into the nucleus at the
nonpermissive temperature. Moreover, when Ran is inefficiently imported
into the nucleus, cells arrest in G2 in a
MAD2 checkpoint-dependent manner. These findings
demonstrate that NTF2 is required to transport Ran into the nucleus in
vivo. Furthermore, we present data that suggest that depletion of
nuclear Ran triggers a spindle-assembly checkpoint-dependent cell cycle arrest.
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