期刊
MOLECULAR AND CELLULAR BIOLOGY
卷 25, 期 2, 页码 728-739出版社
AMER SOC MICROBIOLOGY
DOI: 10.1128/MCB.25.2.728-739.2005
关键词
-
资金
- NCI NIH HHS [N01-CO-12400, N01CO12400] Funding Source: Medline
- DIVISION OF BASIC SCIENCES - NCI [Z01BC010397] Funding Source: NIH RePORTER
- DIVISION OF CLINICAL SCIENCES - NCI [Z01SC010387] Funding Source: NIH RePORTER
- NATIONAL CANCER INSTITUTE [Z01BC010750] Funding Source: NIH RePORTER
The nucleocytoplasmic transport receptor CRM1 mediates the export of macromolecules from the nucleus to the cytoplasm by forming a ternary complex with a cargo molecule and RanGTP. The in vivo mechanism of CRM1 export complex formation and its mobility throughout the nucleus have not been fully elucidated. More information is required to fully understand complex formation and the dynamics of CRM1-cargo-RanGTP complexes in space and time. We demonstrate true molecular interaction of CRM1 with its Rev cargo in living cells by using fluorescence resonance energy transfer (FRET). Interestingly, we found that the inhibitory effect of leptomycin B on this CRM1-cargo interaction is Ran dependent. Using fluorescence recovery after photobleaching (FRAP), we show that CRM1 moves at rates similar to that of free green fluorescent protein in the nucleoplasm. A slower mobility was detected on the nuclear membrane, consistent with known CRM1 interactions with nuclear pores. Based on these data, we propose an in vivo model in which CRM1 roams through the nucleus in search of high-affinity binding sites. CRM1 is able to bind Rev cargo in the nucleolus, and upon RanGTP binding a functional export complex is produced that is exported to the cytoplasm.
作者
我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。
推荐
暂无数据