Shumaker Dale K, Solimando Liliana, Sengupta Kaushik, Shimi Takeshi, Adam Stephen A, Grunwald Antje, Strelkov Sergei V, Aebi Ueli, Cardoso M Cristina, Goldman Robert D
Department of Cell and Molecular Biology, Feinberg School of Medicine, Northwestern University, Chicago, IL 60611, USA.
J Cell Biol. 2008 Apr 21;181(2):269-80. doi: 10.1083/jcb.200708155.
This study provides insights into the role of nuclear lamins in DNA replication. Our data demonstrate that the Ig-fold motif located in the lamin C terminus binds directly to proliferating cell nuclear antigen (PCNA), the processivity factor necessary for the chain elongation phase of DNA replication. We find that the introduction of a mutation in the Ig-fold, which alters its structure and causes human muscular dystrophy, inhibits PCNA binding. Studies of nuclear assembly and DNA replication show that lamins, PCNA, and chromatin are closely associated in situ. Exposure of replicating nuclei to an excess of the lamin domain containing the Ig-fold inhibits DNA replication in a concentration-dependent fashion. This inhibitory effect is significantly diminished in nuclei exposed to the same domain bearing the Ig-fold mutation. Using the crystal structures of the lamin Ig-fold and PCNA, molecular docking simulations suggest probable interaction sites. These findings also provide insights into the mechanisms underlying the numerous disease-causing mutations located within the lamin Ig-fold.
本研究深入探讨了核纤层蛋白在DNA复制中的作用。我们的数据表明,位于核纤层蛋白C末端的免疫球蛋白折叠基序直接与增殖细胞核抗原(PCNA)结合,PCNA是DNA复制链延伸阶段所需的持续性因子。我们发现,在免疫球蛋白折叠中引入突变会改变其结构并导致人类肌肉萎缩症,该突变会抑制PCNA结合。对核组装和DNA复制的研究表明,核纤层蛋白、PCNA和染色质在原位紧密相关。将复制细胞核暴露于过量的含有免疫球蛋白折叠的核纤层蛋白结构域会以浓度依赖的方式抑制DNA复制。在暴露于带有免疫球蛋白折叠突变的相同结构域的细胞核中,这种抑制作用会显著减弱。利用核纤层蛋白免疫球蛋白折叠和PCNA的晶体结构,分子对接模拟揭示了可能的相互作用位点。这些发现还为深入了解位于核纤层蛋白免疫球蛋白折叠内的众多致病突变的潜在机制提供了思路。