International Journal of Legal Medicine

, Volume 118, Issue 5, pp 274–281 | Cite as

Different methods to determine length heteroplasmy within the mitochondrial control region

  • Sabine Lutz-Bonengel
  • Timo Sänger
  • Stefan Pollak
  • Reinhard Szibor
Original Article

Abstract

The first and second hypervariable regions of the human mitochondrial DNA control region contain two homopolymeric stretches of cytosine (nt 16184–16193 and nt 303–315, respectively). According to the Cambridge reference sequence these homopolymeric stretches are interrupted by thymine (T), at positions 16189 and 310, respectively. Monotonous runs of the same base have been suggested to be hot spots for mutations, probably caused by replication slippage, resulting in length heteroplasmy. This paper describes a rapid method based on restriction cleavage of labelled PCR products encompassing the homopolymeric tract in HVII to quantify the relative proportions of different length variants present in an individual. To compare the accuracy of this method, cloned PCR products from several heteroplasmic individuals have been additionally sequenced.

Keywords

mtDNA Homopolymeric tract Length heteroplasmy 

References

  1. Anderson S, Bankier AT, Barrell BG et al. (1981) Sequence and organization of the human mitochondrial genome. Nature 290:457–464PubMedGoogle Scholar
  2. Bär W, Brinkmann B, Budowle B et al. (2000) DNA Commission of the International Society for Forensic Genetics: guidelines for mitochondrial DNA typing. Int J Legal Med 113:193–196CrossRefPubMedGoogle Scholar
  3. Bendall KE, Sykes BC (1995) Length heteroplasmy in the first hypervariable segment of the human mtDNA control region. Am J Hum Genet 57:248–256PubMedGoogle Scholar
  4. Brandstätter A, Niederstätter H, Parson W (2004) Monitoring the inheritance of heteroplasmy by computer-assisted detection of mixed basecalls in the entire human mitochondrial DNA control region. Int J Legal Med 118:47–54CrossRefPubMedGoogle Scholar
  5. Gocke CD, Benko FA, Rogan PK (1998) Transmission of mitochondrial DNA heteroplasmy in normal pedigrees. Hum Genet 102:182–186PubMedGoogle Scholar
  6. Greenberg BD, Newbold JE, Sugino A (1983) Intraspecific nucleotide sequence variability surrounding the origin of replication in human mitochondrial DNA. Gene 21:33–49CrossRefPubMedGoogle Scholar
  7. Hauswirth WW, Clayton DA (1985) Length heterogeneity of a conserved displacement-loop sequence in human mitochondrial DNA. Nucleic Acids Res 13:8093–8104PubMedGoogle Scholar
  8. Hauswirth WW, Van de Walle MJ, Laipis PJ, Olivo PD (1984) Heterogeneous mitochondrial DNA D-loop sequences in bovine tissue. Cell 37:1001–1007PubMedGoogle Scholar
  9. Holland MM, Fisher DL, Roby RK, Ruderman J, Bryson C, Weedn VW (1995) Mitochondrial DNA sequence analysis of human remains. Crime Lab Digest 22:109–115Google Scholar
  10. Hühne J, Pfeiffer H, Brinkmann B (1998) Heteroplasmic substitutions in the mitochondrial DNA control region in mother and child samples. J Legal Med 112:27–30CrossRefGoogle Scholar
  11. Imaizumi K, Parsons TJ, Yoshino M, Holland MM (2002) A new database of mitochondrial DNA hypervariable regions I and II sequences from 162 Japanese individuals. Int J Legal Med 116:68–73PubMedGoogle Scholar
  12. Lee HY, Chung U, Yoo JE, Park MJ, Shin KJ (2004) Quantitative and qualitative profiling of mitochondrial DNA length heteroplasmy. Electrophoresis 25:28–34CrossRefPubMedGoogle Scholar
  13. Lutz S, Weisser HJ, Heizmann J, Pollak S (1998) Location and frequency of polymorphic positions in the mtDNA control region of individuals from Germany. Int J Legal Med 111:67–77, erratum Int J Legal Med (1998) 111:286, Int J Legal Med (1999) 112:145–150Google Scholar
  14. Lutz S, Weisser HJ, Heizmann J, Pollak S (2000) Mitochondrial heteroplasmy among maternally related individuals. Int J Legal Med 113:155–161PubMedGoogle Scholar
  15. Parson W, Parsons TJ, Scheithauer R, Holland MM (1998) Population data for 101 Austrian Caucasian mitochondrial DNA d-loop sequences: application of mtDNA sequence analysis to a forensic case. Int J Legal Med 111:124–132PubMedGoogle Scholar
  16. Pfeiffer H, Lutz-Bonengel S, Pollak S, Fimmers R, Baur MP, Brinkmann B (2004) Mitochondrial DNA control region diversity in hairs and body fluids of monozygotic triplets. Int J Legal Med 118:71–74CrossRefPubMedGoogle Scholar
  17. Piercy R, Sullivan KM, Benson N, Gill P (1993) The application of mitochondrial DNA typing to the study of white Caucasian genetic identification. Int J Legal Med 106:85–90PubMedGoogle Scholar
  18. Rasmussen EM, Sorensen E, Eriksen B, Larsen HJ, Morling N (2002) Sequencing strategy of mitochondrial HV1 and HV2 DNA with length heteroplasmy. Forensic Sci Int 129:209–213CrossRefPubMedGoogle Scholar
  19. Stewart JE, Fisher CL, Aagaard PJ, Wilson MR, Isenberg AR, Polanskey D, Pokorak E, DiZinno JA, Budowle B (2001) Length variation in HV2 of the human mitochondrial DNA control region. J Forensic Sci 46:862–870PubMedGoogle Scholar
  20. Szibor R, Michael M (1999) Correct mitochondrial L-strand sequencing after C-stretches. Int J Legal Med 112:348–349CrossRefPubMedGoogle Scholar
  21. Szibor R, Michael M, Spitsyn VA, Plate I, Ginter EK, Krause D (1997) Mitochondrial D-loop 3’ (CA)n repeat polymorphism: optimization of analysis and population data. Electrophoresis 18:2857–2860PubMedGoogle Scholar
  22. Wilson MR, Allard MW, Monson K, Miller KW, Budowle B (2002) Recommendations for consistent treatment of length variants in the human mitochondrial DNA control region. Forensic Sci Int 129:35–42CrossRefPubMedGoogle Scholar
  23. Zupanic Pajnic I, Balazic J, Komel R (2004) Sequence polymorphism of the mitochondrial DNA control region in the Slovenian population. Int J Legal Med 118:1–4CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag 2004

Authors and Affiliations

  • Sabine Lutz-Bonengel
    • 1
  • Timo Sänger
    • 1
  • Stefan Pollak
    • 1
  • Reinhard Szibor
    • 2
  1. 1.Institute of Legal MedicineAlbert Ludwig University Freiburg FreiburgGermany
  2. 2.Institute of Legal MedicineOtto von Guericke University MagdeburgMagdeburgGermany

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