Centre de Recherche en Cancérologie de Lyon, INSERM U1052/CNRS UMR 5286, Lyon, France.
Oncogene. 2012 Mar 22;31(12):1582-91. doi: 10.1038/onc.2011.342. Epub 2011 Aug 22.
LKB1 is a tumor suppressor that is constitutionally mutated in a cancer-prone condition, called Peutz-Jeghers syndrome, as well as somatically inactivated in a sizeable fraction of lung and cervical neoplasms. The LKB1 gene encodes a serine/threonine kinase that associates with the pseudokinase STRAD (STE-20-related pseudokinase) and the scaffolding protein MO25, the formation of this heterotrimeric complex promotes allosteric activation of LKB1. We have previously reported that the molecular chaperone heat shock protein 90 (Hsp90) binds to and stabilizes LKB1. Combining pharmacological studies and RNA interference approaches, we now provide evidence that the co-chaperone Cdc37 participates to the regulation of LKB1 stability. It is known that the Hsp90-Cdc37 complex recognizes a surface within the N-terminal catalytic lobe of client protein kinases. In agreement with this finding, we found that the chaperones Hsp90 and Cdc37 interact with an LKB1 isoform that differs in the C-terminal region, but not with a novel LKB1 variant that lacks a portion of the kinase N-terminal lobe domain. Reconstitution of the two complexes LKB1-STRAD and LKB1-Hsp90-Cdc37 with recombinant proteins revealed that the former is catalytically active whereas the latter is inactive. Furthermore, consistent with a documented repressor function of Hsp90, LKB1 kinase activity was transiently stimulated upon dissociation of Hsp90. Finally, disruption of the LKB1-Hsp90 complex favors the recruitment of both Hsp/Hsc70 and the U-box dependent E3 ubiquitin ligase CHIP (carboxyl terminus of Hsc70-interacting protein) that triggers LKB1 degradation. Taken together, our results establish that the Hsp90-Cdc37 complex controls both the stability and activity of the LKB1 kinase. This study further shows that two chaperone complexes with antagonizing activities, Hsp90-Cdc37 and Hsp/Hsc70-CHIP, finely control the cellular level of LKB1 protein.
LKB1 是一种肿瘤抑制因子,在易患癌症的条件下(称为 Peutz-Jeghers 综合征)发生结构突变,并且在相当一部分肺癌和宫颈癌中发生体细胞失活。LKB1 基因编码一种丝氨酸/苏氨酸激酶,它与拟激酶 STRAD(STE-20 相关的拟激酶)和支架蛋白 MO25 相关联,这种异三聚体复合物的形成促进了 LKB1 的变构激活。我们之前报道过分子伴侣热休克蛋白 90(Hsp90)与 LKB1 结合并稳定 LKB1。通过药理学研究和 RNA 干扰方法,我们现在提供的证据表明,伴侣蛋白 Cdc37 参与 LKB1 稳定性的调节。众所周知,Hsp90-Cdc37 复合物识别客户蛋白激酶的 N 端催化叶内的一个表面。与这一发现一致,我们发现伴侣蛋白 Hsp90 和 Cdc37 与 LKB1 同种型相互作用,该同种型在 C 端区域不同,但与缺乏激酶 N 端叶域部分的新型 LKB1 变体不相互作用。用重组蛋白重建 LKB1-STRAD 和 LKB1-Hsp90-Cdc37 两个复合物,揭示了前者具有催化活性,而后者没有活性。此外,与 Hsp90 的公认的抑制剂功能一致,Hsp90 的解离短暂地刺激了 LKB1 的激酶活性。最后,LKB1-Hsp90 复合物的破坏有利于 HSP/Hsc70 和 U 盒依赖性 E3 泛素连接酶 CHIP(Hsc70 相互作用蛋白的羧基末端)的募集,从而触发 LKB1 的降解。总之,我们的结果表明,Hsp90-Cdc37 复合物控制 LKB1 激酶的稳定性和活性。这项研究进一步表明,两种具有拮抗活性的伴侣复合物,Hsp90-Cdc37 和 HSP/Hsc70-CHIP,精细地控制 LKB1 蛋白的细胞水平。