The MLH1 D132H variant is associated with susceptibility to sporadic colorectal cancer

Article metrics


Most susceptibility to colorectal cancer (CRC) is not accounted for by known risk factors. Because MLH1, MSH2 and MSH6 mutations underlie high-penetrance CRC susceptibility in hereditary nonpolyposis colon cancer (HNPCC), we hypothesized that attenuated alleles might also underlie susceptibility to sporadic CRC. We looked for gene variants associated with HNPCC in Israeli probands with familial CRC unstratified with respect to the microsatellite instability (MSI) phenotype. Association studies identified a new MLH1 variant (415G→C, resulting in the amino acid substitution D132H) in 1.3% of Israeli individuals with CRC self-described as Jewish, Christian and Muslim. MLH1 415C confers clinically significant susceptibility to CRC. In contrast to classic HNPCC, CRCs associated with MLH1 415C usually do not have the MSI defect, which is important for clinical mutation screening. Structural and functional analyses showed that the normal ATPase function of MLH1 is attenuated, but not eliminated, by the MLH1 415G→C mutation. The new MLH1 variant confers a high risk of CRC and identifies a previously unrecognized mechanism in microsatellite-stable tumors. These studies suggest that variants of mismatch repair proteins with attenuated function may account for a higher proportion of susceptibility to sporadic microsatellite-stable CRC than previously assumed.

Access optionsAccess options

Rent or Buy article

Get time limited or full article access on ReadCube.


All prices are NET prices.

Figure 1: Variant allele detection by the HNPCC Chip resequencing array.
Figure 2: Pedigrees of selected carriers of MLH1 415C.
Figure 3: Structural and functional analyses of MLH1 D132H.

Accession codes


Protein Data Bank


  1. 1

    Wijnen, J. et al. Familial endometrial cancer in female carriers of MSH6 germline mutations. Nat. Genet. 23, 142–144 (1999).

  2. 2

    Charames, G.S., Millar, A.L., Pal, T., Narod, S. & Bapat, B. Do MSH6 mutations contribute to double primary cancers of the colorectum and endometrium? Hum. Genet. 107, 623–629 (2000).

  3. 3

    Peterlongo, P. et al. MSH6 germline mutations are rare in colorectal cancer families. Int. J. Cancer 107, 571–579 (2003).

  4. 4

    Szabo, C.I. et al. Are ATM mutations 7271T→G and IVS10-6T→G really high-risk breast cancer-susceptibility alleles? Cancer Res. 64, 840–843 (2004).

  5. 5

    Laken, S.J. et al. Familial colorectal cancer in Ashkenazim due to a hypermutable tract in APC. Nat. Genet. 17, 79–83 (1997).

  6. 6

    de Wind, N. et al. HNPCC-like cancer predisposition in mice through simultaneous loss of Msh3 and Msh6 mismatch-repair protein functions. Nat. Genet. 23, 359–362 (1999).

  7. 7

    Spampinato, C. & Modrich, P. The MutL ATPase is required for mismatch repair. J. Biol. Chem. 275, 9863–9869 (2000).

  8. 8

    Tran, P.T. & Liskay, R.M. Functional studies on the candidate ATPase domains of Saccharomyces cerevisiae MutLalpha. Mol. Cell Biol. 20, 6390–6398 (2000).

  9. 9

    Scholz, B., Rechter, S., Drach, J.C., Townsend, L.B. & Bogner, E. Identification of the ATP-binding site in the terminase subunit pUL56 of human cytomegalovirus. Nucleic Acids Res. 31, 1426–1433 (2003).

  10. 10

    Guarne, A., Junop, M.S. & Yang, W. Structure and function of the N-terminal 40 kDa fragment of human PMS2: a monomeric GHL ATPase. EMBO J. 20, 5521–5531 (2001).

  11. 11

    Liu, T. et al. Missense mutations in hMLH1 associated with colorectal cancer. Hum. Genet. 105, 437–441 (1999).

  12. 12

    Farrington, S.M. et al. Systematic analysis of hMSH2 and hMLH1 in young colon cancer patients and controls. Am. J. Hum. Genet. 63, 749–759 (1998).

  13. 13

    Scartozzi, M. et al. Mutations of hMLH1 and hMSH2 in patients with suspected hereditary nonpolyposis colorectal cancer: correlation with microsatellite instability and abnormalities of mismatch repair protein expression. J. Clin. Oncol. 20, 1203–1208 (2002).

  14. 14

    Samowitz, W.S. et al. Missense mismatch repair gene alterations, microsatellite instability, and hereditary nonpolyposis colorectal cancer. J. Clin. Oncol. 20, 3178–3179 (2002).

  15. 15

    Guerrette, S., Acharya, S. & Fishel, R. The interaction of the human MutL homologues in hereditary nonpolyposis colon cancer. J. Biol. Chem. 274, 6336–6341 (1999).

  16. 16

    Kolodner, R.D. Germline MSH6 mutation in colorectal cancer families. Cancer Res. 59, 5068–5074 (1999).

  17. 17

    Das Gupta, R. & Kolodner, R.D. Novel dominant mutations in Saccharomyces cerevisiae MSH6. Nat. Genet. 24, 53–56 (2000).

  18. 18

    Gazzoli, I. & Kolodner, R.D. Regulation of the human MSH6 gene by the Sp1 transcription factor and alteration of promoter activity and expression by polymorphisms. Mol. Cell. Biol. 23, 7992–8007 (2003).

  19. 19

    Lynch, H.T. & de la Chapelle, A. Hereditary colorectal cancer. N. Engl. J. Med. 348, 919–932 (2003).

  20. 20

    Shiri-Sverdlov, R. et al. Mutational analyses of BRCA1 and BRCA2 in Ashkenazi and non-Ashkenazi Jewish women with familial breast and ovarian cancer. Hum. Mutat. 16, 491–501 (2000).

  21. 21

    Foulkes, W.D. et al. The founder mutation MSH2*1906G→C is an important cause of hereditary nonpolyposis colorectal cancer in the Ashkenazi Jewish population. Am. J. Hum. Genet. 71, 1395–1412 (2002).

  22. 22

    Gruber, S.B. et al. BLM heterozygosity and the risk of colorectal cancer. Science 297, 2013 (2002).

  23. 23

    Greenson, J.K. et al. Phenotype of microsatellite unstable colorectal carcinomas: Well-differentiated and focally mucinous tumors and the absence of dirty necrosis correlate with microsatellite instability. Am. J. Surg. Pathol. 27, 563–570 (2003).

  24. 24

    Hacia, J.G. et al. Two color hybridization analysis using high density oligonucleotide arrays and energy transfer dyes. Nucleic Acids Res. 26, 3865–3866 (1998).

  25. 25

    Kokoris, M. et al. High-throughput SNP genotyping with the Masscode system. Mol. Diagn. 5, 329–340 (2000).

  26. 26

    Boland, C.R. et al. A National Cancer Institute Workshop on microsatellite instability for cancer detection and familial predisposition: development of international criteria for the determination of microsatellite instability in colorectal cancer. Cancer Res. 58, 5248–5257 (1998).

  27. 27

    Jones, T.A., Zou, J.Y., Cowan, S.W. & Kjeldgaard . Improved methods for building protein models in electron density maps and the location of errors in these models. Acta Crystallogr. A 47 (Pt 2), 110–119 (1991).

  28. 28

    Lipkin, S.M., Naar, A.M., Kalla, K.A., Sack, R.A. & Rosenfeld, M.G. Identification of a novel zinc finger protein binding a conserved element critical for Pit-1-dependent growth hormone gene expression. Genes Dev. 7, 1674–1687 (1993).

  29. 29

    Acharya, S., Foster, P.L., Brooks, P. & Fishel, R. The coordinated functions of the E. coli MutS and MutL proteins in mismatch repair. Mol. Cell. 12, 233–246 (2003).

  30. 30

    Quaresima, B. et al. Human mismatch-repair protein MutL homologue 1 (MLH1) interacts with Escherichia coli MutL and MutS in vivo and in vitro: a simple genetic system to assay MLH1 function. Biochem. J. 371, 183–189 (2003).

Download references


We would like to thank J. Nguyen and L. Paxton for technical assistance. This work was supported by an American Cancer Society Research Scholar Grant (to S.M.L.), Public Health Service grants (to S.B.G. and L.S.R.) and the Ravitz Foundation (S.B.G.).

Author information

Correspondence to Steven M Lipkin or Stephen B Gruber.

Ethics declarations

Competing interests

S.F. is the CEO of Affymetrix Corporation.

Supplementary information

Supplementary Table 1

MLH1, MSH2 and MSH6 exon and peri-exonic variant allels identified in Israeli familial CRC probands. (PDF 8 kb)

Supplementary Table 2

Israeli population based CRC allele frequencies of MLH1, MSH2 and MSH6 variants. (PDF 3 kb)

Supplementary Table 3

Pathological characteristics of CRCs from MLH1 D132H carriers. (PDF 2 kb)

Supplementary Table 4

Microsatellite instability results for subjects carrying the MLH1 D132H variant allele with available tumors. (PDF 2 kb)

Rights and permissions

Reprints and Permissions

About this article

Further reading