Mitogen-Activated Protein/Extracellular Signal-Regulated Kinase Kinase 1act/Tubulin Interaction Is an Important Determinant of Mitotic Stability in Cultured HT1080 Human Fibrosarcoma Cells
Mitogen-Activated Protein/Extracellular Signal-Regulated Kinase Kinase 1act/Tubulin Interaction Is an Important Determinant of Mitotic Stability in Cultured HT1080 Human Fibrosarcoma Cells
复制标题
DOI：
10.1158/0008-5472.can-09-4490
复制
发表时间：
2010-07-15
期刊：
影响因子：
11.2
通讯作者：
Stanbridge, Eric J.
中科院分区：
文献类型：
--
作者：
Cao, Jia-ning;Shafee, Norazizah;Stanbridge, Eric J.
丝裂原活化蛋白激酶(MAPK)通路的激活在肿瘤细胞转化中起重要作用。利用蛋白质组学方法，我们确定α微管蛋白和β微管蛋白是与激活的MAP/细胞外信号调节激酶1(MEK1)相互作用的蛋白质，MEK1是一种中央MAPK调节激酶。共聚焦分析显示，在不同的HT1080人纤维肉瘤细胞中，MEK1-微管蛋白共定位的时空控制在有丝分裂纺锤体中最为显著。多肽阵列确定了MEK1表面的正电氨基酸R108、R113、R160和K157在微管蛋白相互作用中的关键作用。激活的MEK1过表达导致纺锤体排列、染色体分离和倍性缺陷。相反，染色体多倍体在激活的MEK1突变体(R108A，R113A)的存在下减少，该突变体破坏了与微管蛋白的相互作用。我们的发现表明激活的MEK1-微管蛋白信号在纺锤体组织和染色体不稳定中的重要性。巨蟹座；70(14)；6004-14。(C)2010年AACR。
Activation of the mitogen-activated protein kinase (MAPK) pathway plays a major role in neoplastic cell transformation. Using a proteomics approach, we identified alpha tubulin and beta tubulin as proteins that interact with activated MAP/extracellular signal-regulated kinase kinase 1 (MEK1), a central MAPK regulatory kinase. Confocal analysis revealed spatiotemporal control of MEK1-tubulin colocalization that was most prominent in the mitotic spindle apparatus in variant HT1080 human fibrosarcoma cells. Peptide arrays identified the critical role of positively charged amino acids R108, R113, R160, and K157 on the surface of MEK1 for tubulin interaction. Overexpression of activated MEK1 caused defects in spindle arrangement, chromosome segregation, and ploidy. In contrast, chromosome polyploidy was reduced in the presence of an activated MEK1 mutant (R108A, R113A) that disrupted interactions with tubulin. Our findings indicate the importance of signaling by activated MEK1-tubulin in spindle organization and chromosomal instability. Cancer Res; 70(14); 6004-14. (C) 2010 AACR.