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. 1996 Nov;16(11):6516–6523. doi: 10.1128/mcb.16.11.6516

Loss of heterozygosity and base substitution at the APRT locus in mismatch-repair-proficient and -deficient colorectal carcinoma cell lines.

G Phear 1, N P Bhattacharyya 1, M Meuth 1
PMCID: PMC231653  PMID: 8887680

Abstract

We determined the nature of mutations occurring at the autosomal APRT locus in mismatch-repair-proficient and -deficient colorectal carcinoma cell lines. The analysis of mutations that result in APRT deficiency in a mismatch-repair-deficient strain of DLD-1 heterozygous for this locus enabled us to measure the rate of loss of the wild-type gene through deletion, recombination, or gene conversion as well as the rate of point mutation. The overall rate of mutation at the APRT locus in DLD-1 was elevated 100-fold compared with the mismatch-repair-proficient colorectal carcinoma cell line SW620. Loss of heterozygosity (LOH) at APRT accounted for only 4 to 9% of mutant strains derived from DLD-1, indicating a rate for these types of events of 4 x 10(-7) to 9 x 10(-7). In SW620 the rate of LOH at APRT was about 10-fold higher. LOH was not found at polymorphic markers within the same chromosome subband as APRT, indicating that only a limited portion of the chromosome was affected by these alterations. Chromosome painting of SWS620 mutants revealed that the loss of APRT occurred together with a substantial portion of the long arm of chromosome 16. Differences in the nature of base substitutions at APRT (e.g., the proportion of mutations resulting from transitions or transversions) in these tumor cell lines were also detected. There was also an important similarity---the presence of a mutant APRT gene with multiple base substitutions that may be the result of some sort of error-prone DNA synthesis.

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Selected References

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  1. Alani E., Reenan R. A., Kolodner R. D. Interaction between mismatch repair and genetic recombination in Saccharomyces cerevisiae. Genetics. 1994 May;137(1):19–39. doi: 10.1093/genetics/137.1.19. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Benjamin M. B., Little J. B. X rays induce interallelic homologous recombination at the human thymidine kinase gene. Mol Cell Biol. 1992 Jun;12(6):2730–2738. doi: 10.1128/mcb.12.6.2730. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bhattacharyya N. P., Ganesh A., Phear G., Richards B., Skandalis A., Meuth M. Molecular analysis of mutations in mutator colorectal carcinoma cell lines. Hum Mol Genet. 1995 Nov;4(11):2057–2064. doi: 10.1093/hmg/4.11.2057. [DOI] [PubMed] [Google Scholar]
  4. Bhattacharyya N. P., Skandalis A., Ganesh A., Groden J., Meuth M. Mutator phenotypes in human colorectal carcinoma cell lines. Proc Natl Acad Sci U S A. 1994 Jul 5;91(14):6319–6323. doi: 10.1073/pnas.91.14.6319. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Boyer J. C., Umar A., Risinger J. I., Lipford J. R., Kane M., Yin S., Barrett J. C., Kolodner R. D., Kunkel T. A. Microsatellite instability, mismatch repair deficiency, and genetic defects in human cancer cell lines. Cancer Res. 1995 Dec 15;55(24):6063–6070. [PubMed] [Google Scholar]
  6. Bronner C. E., Baker S. M., Morrison P. T., Warren G., Smith L. G., Lescoe M. K., Kane M., Earabino C., Lipford J., Lindblom A. Mutation in the DNA mismatch repair gene homologue hMLH1 is associated with hereditary non-polyposis colon cancer. Nature. 1994 Mar 17;368(6468):258–261. doi: 10.1038/368258a0. [DOI] [PubMed] [Google Scholar]
  7. Capizzi R. L., Jameson J. W. A table for the estimation of the spontaneous mutation rate of cells in culture. Mutat Res. 1973 Jan;17(1):147–148. doi: 10.1016/0027-5107(73)90265-0. [DOI] [PubMed] [Google Scholar]
  8. Drummond J. T., Li G. M., Longley M. J., Modrich P. Isolation of an hMSH2-p160 heterodimer that restores DNA mismatch repair to tumor cells. Science. 1995 Jun 30;268(5219):1909–1912. doi: 10.1126/science.7604264. [DOI] [PubMed] [Google Scholar]
  9. Eshleman J. R., Lang E. Z., Bowerfind G. K., Parsons R., Vogelstein B., Willson J. K., Veigl M. L., Sedwick W. D., Markowitz S. D. Increased mutation rate at the hprt locus accompanies microsatellite instability in colon cancer. Oncogene. 1995 Jan 5;10(1):33–37. [PubMed] [Google Scholar]
  10. Fishel R., Kolodner R. D. Identification of mismatch repair genes and their role in the development of cancer. Curr Opin Genet Dev. 1995 Jun;5(3):382–395. doi: 10.1016/0959-437x(95)80055-7. [DOI] [PubMed] [Google Scholar]
  11. Fishel R., Lescoe M. K., Rao M. R., Copeland N. G., Jenkins N. A., Garber J., Kane M., Kolodner R. The human mutator gene homolog MSH2 and its association with hereditary nonpolyposis colon cancer. Cell. 1993 Dec 3;75(5):1027–1038. doi: 10.1016/0092-8674(93)90546-3. [DOI] [PubMed] [Google Scholar]
  12. Fougerousse F., Meloni R., Roudaut C., Beckmann J. S. Dinucleotide repeat polymorphism at the human hemoglobin alpha-1 pseudo-gene (HBAP1). Nucleic Acids Res. 1992 Mar 11;20(5):1165–1165. [PMC free article] [PubMed] [Google Scholar]
  13. Fukuchi K., Martin G. M., Monnat R. J., Jr Mutator phenotype of Werner syndrome is characterized by extensive deletions. Proc Natl Acad Sci U S A. 1989 Aug;86(15):5893–5897. doi: 10.1073/pnas.86.15.5893. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Harwood J., Tachibana A., Davis R., Bhattacharyya N. P., Meuth M. High rate of multilocus deletion in a human tumor cell line. Hum Mol Genet. 1993 Feb;2(2):165–171. doi: 10.1093/hmg/2.2.165. [DOI] [PubMed] [Google Scholar]
  15. Harwood J., Tachibana A., Meuth M. Multiple dispersed spontaneous mutations: a novel pathway of mutation in a malignant human cell line. Mol Cell Biol. 1991 Jun;11(6):3163–3170. doi: 10.1128/mcb.11.6.3163. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Hidaka Y., Tarlé S. A., O'Toole T. E., Kelley W. N., Palella T. D. Nucleotide sequence of the human APRT gene. Nucleic Acids Res. 1987 Nov 11;15(21):9086–9086. doi: 10.1093/nar/15.21.9086. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Kat A., Thilly W. G., Fang W. H., Longley M. J., Li G. M., Modrich P. An alkylation-tolerant, mutator human cell line is deficient in strand-specific mismatch repair. Proc Natl Acad Sci U S A. 1993 Jul 15;90(14):6424–6428. doi: 10.1073/pnas.90.14.6424. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Leach F. S., Nicolaides N. C., Papadopoulos N., Liu B., Jen J., Parsons R., Peltomäki P., Sistonen P., Aaltonen L. A., Nyström-Lahti M. Mutations of a mutS homolog in hereditary nonpolyposis colorectal cancer. Cell. 1993 Dec 17;75(6):1215–1225. doi: 10.1016/0092-8674(93)90330-s. [DOI] [PubMed] [Google Scholar]
  19. Monnat R. J., Jr, Hackmann A. F., Chiaverotti T. A. Nucleotide sequence analysis of human hypoxanthine phosphoribosyltransferase (HPRT) gene deletions. Genomics. 1992 Jul;13(3):777–787. doi: 10.1016/0888-7543(92)90153-j. [DOI] [PubMed] [Google Scholar]
  20. Nicolaides N. C., Papadopoulos N., Liu B., Wei Y. F., Carter K. C., Ruben S. M., Rosen C. A., Haseltine W. A., Fleischmann R. D., Fraser C. M. Mutations of two PMS homologues in hereditary nonpolyposis colon cancer. Nature. 1994 Sep 1;371(6492):75–80. doi: 10.1038/371075a0. [DOI] [PubMed] [Google Scholar]
  21. Palombo F., Gallinari P., Iaccarino I., Lettieri T., Hughes M., D'Arrigo A., Truong O., Hsuan J. J., Jiricny J. GTBP, a 160-kilodalton protein essential for mismatch-binding activity in human cells. Science. 1995 Jun 30;268(5219):1912–1914. doi: 10.1126/science.7604265. [DOI] [PubMed] [Google Scholar]
  22. Papadopoulos N., Nicolaides N. C., Liu B., Parsons R., Lengauer C., Palombo F., D'Arrigo A., Markowitz S., Willson J. K., Kinzler K. W. Mutations of GTBP in genetically unstable cells. Science. 1995 Jun 30;268(5219):1915–1917. doi: 10.1126/science.7604266. [DOI] [PubMed] [Google Scholar]
  23. Parsons R., Li G. M., Longley M. J., Fang W. H., Papadopoulos N., Jen J., de la Chapelle A., Kinzler K. W., Vogelstein B., Modrich P. Hypermutability and mismatch repair deficiency in RER+ tumor cells. Cell. 1993 Dec 17;75(6):1227–1236. doi: 10.1016/0092-8674(93)90331-j. [DOI] [PubMed] [Google Scholar]
  24. Radman M., Wagner R. Mismatch recognition in chromosomal interactions and speciation. Chromosoma. 1993 Jun;102(6):369–373. doi: 10.1007/BF00360400. [DOI] [PubMed] [Google Scholar]
  25. Reenan R. A., Kolodner R. D. Characterization of insertion mutations in the Saccharomyces cerevisiae MSH1 and MSH2 genes: evidence for separate mitochondrial and nuclear functions. Genetics. 1992 Dec;132(4):975–985. doi: 10.1093/genetics/132.4.975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Schaaper R. M., Dunn R. L. Spectra of spontaneous mutations in Escherichia coli strains defective in mismatch correction: the nature of in vivo DNA replication errors. Proc Natl Acad Sci U S A. 1987 Sep;84(17):6220–6224. doi: 10.1073/pnas.84.17.6220. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Selva E. M., New L., Crouse G. F., Lahue R. S. Mismatch correction acts as a barrier to homeologous recombination in Saccharomyces cerevisiae. Genetics. 1995 Mar;139(3):1175–1188. doi: 10.1093/genetics/139.3.1175. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Shibata D., Peinado M. A., Ionov Y., Malkhosyan S., Perucho M. Genomic instability in repeated sequences is an early somatic event in colorectal tumorigenesis that persists after transformation. Nat Genet. 1994 Mar;6(3):273–281. doi: 10.1038/ng0394-273. [DOI] [PubMed] [Google Scholar]
  29. Thibodeau S. N., Bren G., Schaid D. Microsatellite instability in cancer of the proximal colon. Science. 1993 May 7;260(5109):816–819. doi: 10.1126/science.8484122. [DOI] [PubMed] [Google Scholar]
  30. da Costa L. T., Liu B., el-Deiry W., Hamilton S. R., Kinzler K. W., Vogelstein B., Markowitz S., Willson J. K., de la Chapelle A., Downey K. M. Polymerase delta variants in RER colorectal tumours. Nat Genet. 1995 Jan;9(1):10–11. doi: 10.1038/ng0195-10. [DOI] [PubMed] [Google Scholar]
  31. de la Chapelle A., Peltomäki P. Genetics of hereditary colon cancer. Annu Rev Genet. 1995;29:329–348. doi: 10.1146/annurev.ge.29.120195.001553. [DOI] [PubMed] [Google Scholar]

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