YAP1 is involved in mesothelioma development and negatively regulated by Merlin through phosphorylation.
YAP1 is involved in mesothelioma development and negatively regulated by Merlin through phosphorylation.
复制标题
DOI：
10.1093/carcin/bgn200
复制
发表时间：
2008-11
期刊：
影响因子：
4.7
通讯作者：
T. Yokoyama;H. Osada;H. Murakami;Y. Tatematsu;T. Taniguchi;Y. Kondo;Y. Yatabe;Y. Hasegawa;K. Shimokata;Y. Horio;T. Hida;Y. Sekido
中科院分区：
文献类型：
--
作者：
T. Yokoyama;H. Osada;H. Murakami;Y. Tatematsu;T. Taniguchi;Y. Kondo;Y. Yatabe;Y. Hasegawa;K. Shimokata;Y. Horio;T. Hida;Y. Sekido
我们先前报道了细菌人工染色体阵列综合基因组杂交的恶性胸膜间皮瘤（MPMs），包括两例高水平扩增的11 q22位点的结果。在这项研究中，我们发现，YAP 1基因编码的转录辅激活因子被定位在这一扩增区域，并在这两种情况下过表达，这表明它作为一个候选癌基因在这一地区。我们分析了YAP 1在MPM增殖中的参与，以及其与由神经纤维瘤病2型（NF 2）肿瘤抑制基因编码的Merlin的功能和物理相互作用，该基因在MPM中经常突变。YAP 1-RNA干扰抑制了NF 2纯合缺失的间皮瘤细胞系NCI-H290的生长，可能是通过细胞周期阻滞和凋亡诱导，而YAP 1转染促进了永生化间皮瘤细胞系MeT-5A的生长。我们还发现，NF 2到NCI-H290中的引入诱导了YAP 1的丝氨酸127处的磷酸化，这伴随着YAP 1的核定位的减少，而YAP 1的S127 A突变体的核定位不受影响。此外，免疫沉淀和体外拉下试验的结果表明Merlin和YAP 1之间存在物理相互作用。这些结果表明，YAP 1参与间皮细胞的生长和YAP 1的转录辅激活因子的活性是功能性抑制梅林通过诱导磷酸化和细胞质滞留的YAP 1。这是第一个报告的负调控信号从梅林YAP 1在哺乳动物细胞。未来对MPM中YAP 1转录靶点的研究可能会揭示MPM发展的分子机制，并导致新的治疗策略。
We previously reported the results of bacterial artificial chromosome array comprehensive genomic hybridization of malignant pleural mesotheliomas (MPMs), including two cases with high-level amplification in the 11q22 locus. In this study, we found that the YAP1 gene encoding a transcriptional coactivator was localized in this amplified region and overexpressed in both cases, suggesting it as a candidate oncogene in this region. We analyzed the involvement of YAP1 in MPM proliferation, as well as its functional and physical interaction with Merlin encoded by the neurofibromatosis type 2 (NF2) tumor suppressor gene, which is frequently mutated in MPMs. YAP1-RNA interference suppressed growth of a mesothelioma cell line NCI-H290 with NF2 homozygous deletion, probably through cell-cycle arrest and apoptosis induction, whereas YAP1 transfection promoted the growth of MeT-5A, an immortalized mesothelial cell line. We also found that the introduction of NF2 into NCI-H290 induced phosphorylation at serine 127 of YAP1, which was accompanied by reduction of nuclear localization of YAP1, whereas nuclear localization of a YAP1 S 127A mutant was not affected. Furthermore, results of immunoprecipitation and in vitro pull-down assays indicated a physical interaction between Merlin and YAP1. These results suggest that YAP1 is involved in mesothelial cell growth and that the transcriptional coactivator activity of YAP1 is functionally inhibited by Merlin through the induction of phosphorylation and cytoplasmic retention of YAP1. This is the first report of negative regulatory signaling from Merlin to YAP1 in mammalian cells. Future studies of transcriptional targets of YAP1 in MPMs may shed light on the molecular mechanisms of MPM development and lead to new therapeutic strategies.