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酵母中类AP-1转录因子Yap1p的核输入由转运受体Pse1p介导,且该输入步骤不受氧化应激的影响。

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.

作者信息

Isoyama T, Murayama A, Nomoto A, Kuge S

机构信息

Department of Microbiology, Graduate School of Medicine, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo, 113-0033, Japan.

出版信息

J Biol Chem. 2001 Jun 15;276(24):21863-9. doi: 10.1074/jbc.M009258200. Epub 2001 Mar 23.

Abstract

The yeast AP-1-like transcription factor, Yap1p, is essential for the oxidative stress response in budding yeast. Yap1p is located predominantly in the cytoplasm; however, upon imposition of oxidative stress, Yap1p concentrates in the nucleus and activates target genes. Yap1p is constitutively transported 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 determined whether the import step is affected by oxidative stress. The nuclear accumulation of Yap1p required the activity of the small GTPase, Ran/Gsp1p. Under conditions in pse1-1 cells carrying a temperature-sensitive mutation of the importin beta family member PSE1/KAP121, nuclear translocation of Yap1p was inhibited dramatically. In an in vitro assay, we showed that Yap1p could directly bind to Pse1p and that this interaction was dissociated by Ran-GTP. These results indicate that Pse1p is the nuclear import receptor for Yap1p. In addition to Pse1p, we suggest that Kap123p, which is homologous to Pse1p, has a minor effect on the nuclear import of Yap1p. Furthermore, we identified the nuclear localization signal of Yap1p and demonstrated that the nuclear import of Yap1p was not affected by oxidative stress.

摘要

酵母中类似AP-1的转录因子Yap1p对于芽殖酵母的氧化应激反应至关重要。Yap1p主要位于细胞质中;然而,在施加氧化应激时,Yap1p会集中在细胞核中并激活靶基因。Yap1p持续地进出细胞核。氧化应激会抑制Crm1p/Xpo1p依赖的核输出步骤,导致Yap1p在细胞核中积累。在本研究中,我们研究了Yap1p核输入的机制,并确定输入步骤是否受氧化应激影响。Yap1p在细胞核中的积累需要小GTP酶Ran/Gsp1p的活性。在携带输入蛋白β家族成员PSE1/KAP121温度敏感突变的pse1-1细胞中,Yap1p的核转位受到显著抑制。在体外实验中,我们表明Yap1p可以直接与Pse1p结合,并且这种相互作用会被Ran-GTP解离。这些结果表明Pse1p是Yap1p的核输入受体。除了Pse1p之外,我们认为与Pse1p同源的Kap123p对Yap1p的核输入有较小影响。此外,我们鉴定了Yap1p的核定位信号,并证明Yap1p的核输入不受氧化应激影响。

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