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J. Biol. Chem., Vol. 269, Issue 40, 24542-24545, 10, 1994
T Tachibana, N Imamoto, H Seino, T Nishimoto and Y Yoneda
The role of RCC1-Ran/TC4 in nuclear protein import was examined in living
cells using a temperature-sensitive RCC1 mutant cell line, tsBN2, and tsBN2
transformed with a RCC1 cDNA lacking the nuclear localization sequence
domain, delta 8-29. Substrate, containing a small number of SV40 T antigen
nuclear localization sequence peptides, injected into the cytoplasm of
tsBN2 cells cultured at the non- permissive temperature of 39.5 degrees C
did not accumulate efficiently in the nucleus. When the same substrate was
injected into the cytoplasm of heterokaryons of tsBN2 and wild type BHK21
cells, import efficiency into the tsBN2 nuclei was not restored. Import
into the BHK21 nuclei gradually decreased after fusion. In contrast, import
efficiency into tsBN2 nuclei gradually recovered after fusion with tsBN2
cells transformed with delta 8-29 in which functional RCC1 was diffusely
distributed in both the nuclei and cytoplasm. Substrate did not accumulate
in the nuclei of digitonin-permeabilized tsBN2 cells cultured at 39.5
degrees C even in the presence of normal cytosol. These results suggest
that loss of RCC1 function leads to the decline of import competence of the
nucleus and accumulation of a factor in the cytoplasm that suppresses
nuclear import. These results indicate that the RCC1-Ran/TC4 system may
regulate nuclear import.
Loss of RCC1 leads to suppression of nuclear protein import in living cells
Department of Anatomy and Cell Biology, Osaka University Medical School, Japan.
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