Journal Article

Interactions of the DNA mismatch repair proteins MLH1 and MSH2 with c-MYC and MAX

Gillespie, David AF, Partlin, Mary Mac, Matheson, Elizabeth C, Hall, Andrew G, Crouch, Dorothy H, Brown, Robert, Robinson, Helen, J McCormick, Carol, Durant, Stephen T, Homer, Elizabeth

2003

Overview

MSH2 and MLH1 have a central role in correcting mismatches in DNA occurring during DNA replication and have been implicated in the engagement of apoptosis induced by a number of cytotoxic anticancer agents. The function of MLH1 is not clearly defined, although it is required for mismatch repair (MMR) and engagement of apoptosis after certain types of DNA damage. In order to identify other partners of MLH1 that may be involved in signalling MMR or apoptosis, we used human MLH1 in yeast two-hybrid screens of normal human breast and ovarian cDNA libraries. As well as known partners of MLH1 such as PMS1, MLH3 and MBD4, we identified the carboxy terminus of the human c-MYC proto-oncogene as an interacting sequence. We demonstrate, both in vitro by yeast two-hybrid and GST-fusion pull-down experiments, as well as in vivo by coimmunoprecipitation from human tumour cell extracts, that MLH1 interacts with the c-MYC protein. We further demonstrate that the heterodimeric partner of c-MYC, MAX, interacts with a different MMR protein, MSH2, both in vitro and in vivo. Using an inducible c-MYC-ER™ fusion gene, we show that elevated c-MYC expression leads to an increased HGPRT mutation rate of Rat1 cells and an increase in the number of frameshift mutants at the HGPRT locus. The effect on HGPRT mutation rate is small (2–3-fold), but is consistent with deregulated c-MYC expression partially inhibiting MMR activity.

• Publisher: Nature Publishing Group UK,Nature Publishing,Nature Publishing Group
• Subject: Adaptor Proteins, Signal Transducing / Amino acids / Animals / Antitumor agents / Apoptosis / Base Pair Mismatch / Basic Helix-Loop-Helix Leucine Zipper Transcription Factors / Basic-Leucine Zipper Transcription Factors / Biological and medical sciences / Breast / c-Myc protein / Cancer research / Carrier Proteins / Cell Biology / Cell cycle / Cell physiology / Cell transformation and carcinogenesis. Action of oncogenes and antioncogenes / Cytotoxicity / Deoxyribonucleic acid / DNA / DNA biosynthesis / DNA damage / DNA repair / DNA Repair - physiology / DNA-Binding Proteins - metabolism / Frameshift mutation / Fundamental and applied biological sciences. Psychology / Fusion protein / Human Genetics / Humans / Hypoxanthine Phosphoribosyltransferase - genetics / Internal Medicine / Laboratories / Medical research / Medicine / Medicine & Public Health / Mismatch repair / MLH1 protein / MMR protein / Molecular and cellular biology / MSH2 protein / Mutagenesis - physiology / Mutation / Mutation rates / MutL Protein Homolog 1 / MutS Homolog 2 Protein / Myc protein / Neoplasm Proteins - metabolism / Nuclear Proteins / Oncology / original-paper / Ovaries / Proteins / Proto-Oncogene Proteins - metabolism / Proto-Oncogene Proteins c-myc - biosynthesis / Proto-Oncogene Proteins c-myc - genetics / Proto-Oncogene Proteins c-myc - metabolism / Rats / Transcription Factors / Tumor Cells, Cultured / Tumors / Two-Hybrid System Techniques / Yeast