Decreased Expression of the DNA Mismatch Repair Gene Mlh1 under Hypoxic Stress in Mammalian Cells

Abstract

The hypoxic tumor microenvironment has been shown to contribute to genetic instability. As one possible mechanism for this effect, we report that expression of the DNA mismatch repair (MMR) gene Mlh1 is specifically reduced in mammalian cells under hypoxia, whereas expression of other MMR genes, including Msh2, Msh6, and Pms2, is not altered at the mRNA level. However, levels of the PMS2 protein are reduced, consistent with destabilization of PMS2 in the absence of its heterodimer partner, MLH1. The hypoxia-induced reduction in Mlh1 mRNA was prevented by the histone deacetylase inhibitor trichostatin A, suggesting that hypoxia causes decreased Mlh1 transcription via histone deacetylation. In addition, treatment of cells with the iron chelator desferrioxamine also reduced MLH1 and PMS2 levels, in keeping with low oxygen tension being the stress signal that provokes the altered MMR gene expression. Functional MMR deficiency under hypoxia was detected as induced instability of a (CA)₂₉ dinucleotide repeat and by increased mutagenesis in a chromosomal reporter gene. These results identify a potential new pathway of genetic instability in cancer: hypoxia-induced reduction in the expression of key MMR proteins. In addition, this stress-induced genetic instability may represent a conceptual parallel to the pathway of stationary-phase mutagenesis seen in bacteria.

ACKNOWLEDGMENTS

Valia T. Mihaylova, Ranjit Bindra, and Jianling Yuan contributed equally to this work.

We thank M. Liskay, A. Giaccia, R. Hill, J. Sweasy, A. Perkins, and M. Snyder for helpful discussions and S. Peretz, R. Hickey, K. Wehner, L. Cabral, R. Franklin, and S. J. Baserga for their help. We also thank M. Liskay and B. Vogelstein for providing reagents.

This work was supported by grants from the NIH (ES05775 and CA16038) and the American Cancer Society (RPG-96-129-03-MGO) to P.M.G.