Nuclear import of the yeast AP-1-like transcription factor Yap1p is mediated by transport receptor Pse1p, and this import step is not affected by oxidative stress

The yeast AP-1-like transcription factor, Yap1p, is essential for the oxida tive stress response in budding yeast. Yap1p is located predominantly in th e cytoplasm; however, upon imposition of oxidative stress, Yap1p concentrat es in the nucleus and activates target genes. Yap1p is constitutively trans ported in and out of the nucleus. Oxidative stress inhibits the Crm1p/Xpo1p -dependent nuclear export step, resulting in nuclear accumulation of Yap1p, In this study, we examined the mechanism for Yap1p nuclear import, and det ermined whether the import step is affected by oxidative stress. The nuclea r accumulation of Yap1p required the activity of the small GTPase, Ran/Gsp1 p. Under conditions in pse1-1 cells carrying a temperature-sensitive mutati on of the importin beta family member PSE1/KAP121, nuclear translocation of Yap1p was inhibited dramatically. In an in vitro assay, we showed that Yap 1p could directly bind to Pse1p and that this interaction was dissociated b y Ran-GTP. These results indicate that Pse1p is the nuclear import receptor for Yap1p, In addition to Pse1p, we suggest that Kap123p, which is homolog ous to Pse1p, has a minor effect on the nuclear import of Yap1p. Furthermor e, we identified the nuclear localization signal of Yap1p and demonstrated that the nuclear import of Yap1p was not affected by oxidative stress.