Skip to main content
NCBI home page
Nucleic Acids Research logoLink to Nucleic Acids Research
. 1995 Apr 25;23(8):1419–1425. doi: 10.1093/nar/23.8.1419

Increased frequency of deletions in the mitochondrial genome with age of Caenorhabditis elegans.

S Melov 1, G J Lithgow 1, D R Fischer 1, P M Tedesco 1, T E Johnson 1
PMCID: PMC306871  PMID: 7753635

Abstract

We have developed a long-extension-PCR strategy which amplifies approximately half of the mitochondrial genome (6.3 kb) of Caenorhabditis elegans using an individual worm as target. We analyzed three strains over their life span to assess the number of detectable deletions in the mitochondrial genome. Two of these strains are wild-type for life span while the third is mutant in the age-1 gene, approximately doubling its maximum life span. At the mean life span in wild-type strains, there was a significant difference between the frequency of deletions detected in the mitochondrial genome compared with the mean number of deletions in young animals. In addition, deletions in the mitochondrial genome occur at a significantly lower rate in age-1 mutants as compared with wild type. We cloned and identified the breakpoints of two deletions and found that one of the deletions had a direct repeat of 8 bp at the breakpoint. This is the largest single study (over 900 individual animals) characterizing the frequency of deletions in the mitochondrial genome as a function of age yet carried out.

Full text

PDF
1419

Images in this article

1422Image
on p.1422
1424Image
on p.1424

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Adachi K., Fujiura Y., Mayumi F., Nozuhara A., Sugiu Y., Sakanashi T., Hidaka T., Toshima H. A deletion of mitochondrial DNA in murine doxorubicin-induced cardiotoxicity. Biochem Biophys Res Commun. 1993 Sep 15;195(2):945–951. doi: 10.1006/bbrc.1993.2135. [DOI] [PubMed] [Google Scholar]
  2. Bandy B., Davison A. J. Mitochondrial mutations may increase oxidative stress: implications for carcinogenesis and aging? Free Radic Biol Med. 1990;8(6):523–539. doi: 10.1016/0891-5849(90)90152-9. [DOI] [PubMed] [Google Scholar]
  3. Barnes W. M. PCR amplification of up to 35-kb DNA with high fidelity and high yield from lambda bacteriophage templates. Proc Natl Acad Sci U S A. 1994 Mar 15;91(6):2216–2220. doi: 10.1073/pnas.91.6.2216. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Baumer A., Zhang C., Linnane A. W., Nagley P. Age-related human mtDNA deletions: a heterogeneous set of deletions arising at a single pair of directly repeated sequences. Am J Hum Genet. 1994 Apr;54(4):618–630. [PMC free article] [PubMed] [Google Scholar]
  5. Brown M. D., Wallace D. C. Molecular basis of mitochondrial DNA disease. J Bioenerg Biomembr. 1994 Jun;26(3):273–289. doi: 10.1007/BF00763099. [DOI] [PubMed] [Google Scholar]
  6. Calleja M., Peña P., Ugalde C., Ferreiro C., Marco R., Garesse R. Mitochondrial DNA remains intact during Drosophila aging, but the levels of mitochondrial transcripts are significantly reduced. J Biol Chem. 1993 Sep 5;268(25):18891–18897. [PubMed] [Google Scholar]
  7. Cheng S., Fockler C., Barnes W. M., Higuchi R. Effective amplification of long targets from cloned inserts and human genomic DNA. Proc Natl Acad Sci U S A. 1994 Jun 7;91(12):5695–5699. doi: 10.1073/pnas.91.12.5695. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Cheng S., Higuchi R., Stoneking M. Complete mitochondrial genome amplification. Nat Genet. 1994 Jul;7(3):350–351. doi: 10.1038/ng0794-350. [DOI] [PubMed] [Google Scholar]
  9. Corbisier P., Remacle J. Involvement of mitochondria in cell degeneration. Eur J Cell Biol. 1990 Feb;51(1):173–182. [PubMed] [Google Scholar]
  10. Corral-Debrinski M., Horton T., Lott M. T., Shoffner J. M., Beal M. F., Wallace D. C. Mitochondrial DNA deletions in human brain: regional variability and increase with advanced age. Nat Genet. 1992 Dec;2(4):324–329. doi: 10.1038/ng1292-324. [DOI] [PubMed] [Google Scholar]
  11. Cortopassi G. A., Arnheim N. Detection of a specific mitochondrial DNA deletion in tissues of older humans. Nucleic Acids Res. 1990 Dec 11;18(23):6927–6933. doi: 10.1093/nar/18.23.6927. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Cortopassi G. A., Shibata D., Soong N. W., Arnheim N. A pattern of accumulation of a somatic deletion of mitochondrial DNA in aging human tissues. Proc Natl Acad Sci U S A. 1992 Aug 15;89(16):7370–7374. doi: 10.1073/pnas.89.16.7370. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Friedman D. B., Johnson T. E. Three mutants that extend both mean and maximum life span of the nematode, Caenorhabditis elegans, define the age-1 gene. J Gerontol. 1988 Jul;43(4):B102–B109. doi: 10.1093/geronj/43.4.b102. [DOI] [PubMed] [Google Scholar]
  14. Gadaleta M. N., Rainaldi G., Lezza A. M., Milella F., Fracasso F., Cantatore P. Mitochondrial DNA copy number and mitochondrial DNA deletion in adult and senescent rats. Mutat Res. 1992 Sep;275(3-6):181–193. doi: 10.1016/0921-8734(92)90022-h. [DOI] [PubMed] [Google Scholar]
  15. HARMAN D. Aging: a theory based on free radical and radiation chemistry. J Gerontol. 1956 Jul;11(3):298–300. doi: 10.1093/geronj/11.3.298. [DOI] [PubMed] [Google Scholar]
  16. Harman D. Free radical theory of aging. Mutat Res. 1992 Sep;275(3-6):257–266. doi: 10.1016/0921-8734(92)90030-s. [DOI] [PubMed] [Google Scholar]
  17. Hayashi J., Ohta S., Kikuchi A., Takemitsu M., Goto Y., Nonaka I. Introduction of disease-related mitochondrial DNA deletions into HeLa cells lacking mitochondrial DNA results in mitochondrial dysfunction. Proc Natl Acad Sci U S A. 1991 Dec 1;88(23):10614–10618. doi: 10.1073/pnas.88.23.10614. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Johnson T. E. Increased life-span of age-1 mutants in Caenorhabditis elegans and lower Gompertz rate of aging. Science. 1990 Aug 24;249(4971):908–912. doi: 10.1126/science.2392681. [DOI] [PubMed] [Google Scholar]
  19. Johnson T. E., Lithgow G. J. The search for the genetic basis of aging: the identification of gerontogenes in the nematode Caenorhabditis elegans. J Am Geriatr Soc. 1992 Sep;40(9):936–945. doi: 10.1111/j.1532-5415.1992.tb01993.x. [DOI] [PubMed] [Google Scholar]
  20. Johnson T. E., Tedesco P. M., Lithgow G. J. Comparing mutants, selective breeding, and transgenics in the dissection of aging processes of Caenorhabditis elegans. Genetica. 1993;91(1-3):65–77. doi: 10.1007/BF01435988. [DOI] [PubMed] [Google Scholar]
  21. Larsen P. L. Aging and resistance to oxidative damage in Caenorhabditis elegans. Proc Natl Acad Sci U S A. 1993 Oct 1;90(19):8905–8909. doi: 10.1073/pnas.90.19.8905. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Lee C. M., Chung S. S., Kaczkowski J. M., Weindruch R., Aiken J. M. Multiple mitochondrial DNA deletions associated with age in skeletal muscle of rhesus monkeys. J Gerontol. 1993 Nov;48(6):B201–B205. doi: 10.1093/geronj/48.6.b201. [DOI] [PubMed] [Google Scholar]
  23. Lithgow G. J., White T. M., Hinerfeld D. A., Johnson T. E. Thermotolerance of a long-lived mutant of Caenorhabditis elegans. J Gerontol. 1994 Nov;49(6):B270–B276. doi: 10.1093/geronj/49.6.b270. [DOI] [PubMed] [Google Scholar]
  24. Marton A., Delbecchi L., Bourgaux P. DNA nicking favors PCR recombination. Nucleic Acids Res. 1991 May 11;19(9):2423–2426. doi: 10.1093/nar/19.9.2423. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Melov S., Hertz G. Z., Stormo G. D., Johnson T. E. Detection of deletions in the mitochondrial genome of Caenorhabditis elegans. Nucleic Acids Res. 1994 Mar 25;22(6):1075–1078. doi: 10.1093/nar/22.6.1075. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Miquel J., Economos A. C., Fleming J., Johnson J. E., Jr Mitochondrial role in cell aging. Exp Gerontol. 1980;15(6):575–591. doi: 10.1016/0531-5565(80)90010-8. [DOI] [PubMed] [Google Scholar]
  27. Mita S., Rizzuto R., Moraes C. T., Shanske S., Arnaudo E., Fabrizi G. M., Koga Y., DiMauro S., Schon E. A. Recombination via flanking direct repeats is a major cause of large-scale deletions of human mitochondrial DNA. Nucleic Acids Res. 1990 Feb 11;18(3):561–567. doi: 10.1093/nar/18.3.561. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Ogura R., Sugiyama M., Haramaki N., Hidaka T. Electron spin resonance studies on the mechanism of adriamycin-induced heart mitochondrial damages. Cancer Res. 1991 Jul 1;51(13):3555–3558. [PubMed] [Google Scholar]
  29. Okimoto R., Macfarlane J. L., Clary D. O., Wolstenholme D. R. The mitochondrial genomes of two nematodes, Caenorhabditis elegans and Ascaris suum. Genetics. 1992 Mar;130(3):471–498. doi: 10.1093/genetics/130.3.471. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Orr W. C., Sohal R. S. Extension of life-span by overexpression of superoxide dismutase and catalase in Drosophila melanogaster. Science. 1994 Feb 25;263(5150):1128–1130. doi: 10.1126/science.8108730. [DOI] [PubMed] [Google Scholar]
  31. Pikó L., Hougham A. J., Bulpitt K. J. Studies of sequence heterogeneity of mitochondrial DNA from rat and mouse tissues: evidence for an increased frequency of deletions/additions with aging. Mech Ageing Dev. 1988 Jun;43(3):279–293. doi: 10.1016/0047-6374(88)90037-1. [DOI] [PubMed] [Google Scholar]
  32. Shoffner J. M., Lott M. T., Voljavec A. S., Soueidan S. A., Costigan D. A., Wallace D. C. Spontaneous Kearns-Sayre/chronic external ophthalmoplegia plus syndrome associated with a mitochondrial DNA deletion: a slip-replication model and metabolic therapy. Proc Natl Acad Sci U S A. 1989 Oct;86(20):7952–7956. doi: 10.1073/pnas.86.20.7952. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Soong N. W., Hinton D. R., Cortopassi G., Arnheim N. Mosaicism for a specific somatic mitochondrial DNA mutation in adult human brain. Nat Genet. 1992 Dec;2(4):318–323. doi: 10.1038/ng1292-318. [DOI] [PubMed] [Google Scholar]
  34. Vanfleteren J. R. Oxidative stress and ageing in Caenorhabditis elegans. Biochem J. 1993 Jun 1;292(Pt 2):605–608. doi: 10.1042/bj2920605. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Wei Y. H. Mitochondrial DNA alterations as ageing-associated molecular events. Mutat Res. 1992 Sep;275(3-6):145–155. doi: 10.1016/0921-8734(92)90019-l. [DOI] [PubMed] [Google Scholar]
  36. Zhang C., Baumer A., Maxwell R. J., Linnane A. W., Nagley P. Multiple mitochondrial DNA deletions in an elderly human individual. FEBS Lett. 1992 Feb 3;297(1-2):34–38. doi: 10.1016/0014-5793(92)80321-7. [DOI] [PubMed] [Google Scholar]

Articles from Nucleic Acids Research are provided here courtesy of Oxford University Press

RESOURCES