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Originally published In Press as doi:10.1074/jbc.M908768199 on March 10, 2000

J. Biol. Chem., Vol. 275, Issue 21, 15728-15732, May 26, 2000
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Identification of a Second MutL DNA Mismatch Repair Complex (hPMS1 and hMLH1) in Human Epithelial Cells*

Wai K. LeungDagger §, Jae J. KimDagger , Ling WuDagger , Jorge L. Sepulveda, and Antonia R. SepulvedaDagger ||

From the Departments of Dagger  Medicine and  Pathology, Veterans Affairs Medical Center and Baylor College of Medicine, Houston, Texas 77030

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.

* This study was supported in part by funding from the Texas Gulf Coast Digestive Disease Center, the American Cancer Society (Institutional Research Grant 199A), and the Cutis Hankamer Basic Research Fund (to A. R. S.).The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

§ Partly supported by a Li Po Chun Charitable Trust Fund Overseas Postgraduate Scholarship of Hong Kong.

|| To whom correspondence should be addressed. Antonia R. Sepulveda, M.D. Ph.D., Veterans Affairs Medical Center (111D), 2002 Holcombe Blvd., Houston, TX 77030. Tel.: 713-794-7801; Fax: 713-790-1040; E-mail: asepulv@bcm.tmc.edu.

Copyright © 2000 by The American Society for Biochemistry and Molecular Biology, Inc.
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