Leung W K, Kim J J, Wu L, Sepulveda J L, Sepulveda A R
Departments of Medicine and Pathology, Veterans Affairs Medical Center and Baylor College of Medicine, Houston, Texas 77030, USA.
J Biol Chem. 2000 May 26;275(21):15728-32. doi: 10.1074/jbc.M908768199.
Deficiencies of MutL DNA mismatch repair-complex proteins (hMLH1, hPMS2, and hPMS1) typically result in microsatellite instability in human cancers. We examined the association patterns of MutL proteins in human epithelial cancer cell lines with (HCT-116, N87, SNU-1, and SNU-638) and without microsatellite instability (HeLa, AGS, KATO-III, and SNU-16). The analysis of hMLH1, hPMS2, and hPMS1 was performed using Northern blot, Western blot, and co-immunoprecipitation studies. Our data provide evidence that MutL proteins form two different complexes, MutL-alpha (hPMS2 and hMLH1) and MutL-beta (hPMS1 and hMLH1). Gastric and colorectal cancer cells lines with microsatellite instability lacked detectable hMLH1. Decreased levels of hMLH1 protein were associated with markedly reduced levels of hPMS2 and hPMS1 proteins, but the RNA levels of hPMS1 and hPMS2 were normal. In this study, we describe the association of hPMS1 with hMLH1 as a heterodimer, in human cells. Furthermore, normal levels of hMLH1 protein appear to be important in maintaining normal levels of hPMS1 and hPMS2 proteins.
MutL DNA错配修复复合物蛋白(hMLH1、hPMS2和hPMS1)的缺陷通常会导致人类癌症中的微卫星不稳定性。我们检测了MutL蛋白在有微卫星不稳定性的人类上皮癌细胞系(HCT - 116、N87、SNU - 1和SNU - 638)和无微卫星不稳定性的细胞系(HeLa、AGS、KATO - III和SNU - 16)中的关联模式。使用Northern印迹、Western印迹和免疫共沉淀研究对hMLH1、hPMS2和hPMS1进行了分析。我们的数据表明,MutL蛋白形成两种不同的复合物,MutL - alpha(hPMS2和hMLH1)和MutL - beta(hPMS1和hMLH1)。具有微卫星不稳定性的胃癌和结肠癌细胞系中检测不到hMLH1。hMLH1蛋白水平降低与hPMS2和hPMS1蛋白水平显著降低相关,但hPMS1和hPMS2的RNA水平正常。在本研究中,我们描述了在人类细胞中hPMS1与hMLH1作为异二聚体的关联。此外,正常水平的hMLH1蛋白似乎对维持hPMS1和hPMS2蛋白的正常水平很重要。