Ma Yan, Cai Shang, Lv Quanlong, Jiang Qing, Zhang Quan, Zhai Zhonghe, Zhang Chuanmao
The Key Laboratory of Cell Proliferation and Differentiation of Ministry of Education, The National Key Laboratory of Bio-membrane and Membrane Biotechnology, College of Life Sciences, Peking University, Beijing 100871, China.
J Cell Sci. 2007 Feb 1;120(Pt 3):520-30. doi: 10.1242/jcs.03355.
Lamin B receptor (LBR), a chromatin and lamin binding protein in the inner nuclear membrane, has been proposed to play a vital role in nuclear envelope (NE) assembly. But the specific role for LBR in NE assembly remains unknown. In the present study, we show that overexpression of LBR causes membrane overproduction, inducing NE invagination and membrane stack formation, and that these processes require the transmembrane domain of LBR. Biochemical analysis shows that the N-terminal domain of LBR directly interacts with importin beta in a Ran sensitive and importin alpha independent manner. Using an in vitro NE assembly assay, we also demonstrate that blocking full length LBR binding sites on importin beta, by the addition of the LBR N-terminal domain inhibits the recruitment of LBR-containing vesicles to importin beta- or Ran-coated beads to form NE structure. Our results suggest that LBR is recruited to chromatin through direct interaction with importin beta to contribute to the fusion of membrane vesicles and formation of the NE.
核纤层蛋白B受体(LBR)是内核膜中的一种染色质和核纤层结合蛋白,有人提出它在核膜(NE)组装中起着至关重要的作用。但LBR在NE组装中的具体作用仍不清楚。在本研究中,我们发现LBR的过表达会导致膜过度产生,诱导NE内陷和膜堆叠形成,并且这些过程需要LBR的跨膜结构域。生化分析表明,LBR的N端结构域以一种对Ran敏感且不依赖于输入蛋白α的方式直接与输入蛋白β相互作用。使用体外NE组装试验,我们还证明,通过添加LBR N端结构域来阻断输入蛋白β上的全长LBR结合位点,会抑制含LBR的囊泡募集到输入蛋白β或Ran包被的珠子上以形成NE结构。我们的结果表明,LBR通过与输入蛋白β的直接相互作用被募集到染色质上,从而有助于膜囊泡的融合和NE的形成。