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. 1998 Apr;148(4):1763–1776. doi: 10.1093/genetics/148.4.1763

Functions of the high mobility group protein, Abf2p, in mitochondrial DNA segregation, recombination and copy number in Saccharomyces cerevisiae.

O Zelenaya-Troitskaya 1, S M Newman 1, K Okamoto 1, P S Perlman 1, R A Butow 1
PMCID: PMC1460092  PMID: 9581629

Abstract

Previous studies have established that the mitochondrial high mobility group (HMG) protein, Abf2p, of Saccharomyces cerevisiae influences the stability of wild-type (rho+) mitochondrial DNA (mtDNA) and plays an important role in mtDNA organization. Here we report new functions for Abf2p in mtDNA transactions. We find that in homozygous deltaabf2 crosses, the pattern of sorting of mtDNA and mitochondrial matrix protein is altered, and mtDNA recombination is suppressed relative to homozygous ABF2 crosses. Although Abf2p is known to be required for the maintenance of mtDNA in rho+ cells growing on rich dextrose medium, we find that it is not required for the maintenance of mtDNA in p cells grown on the same medium. The content of both rho+ and rho- mtDNAs is increased in cells by 50-150% by moderate (two- to threefold) increases in the ABF2 copy number, suggesting that Abf2p plays a role in mtDNA copy control. Overproduction of Abf2p by > or = 10-fold from an ABF2 gene placed under control of the GAL1 promoter, however, leads to a rapid loss of rho+ mtDNA and a quantitative conversion of rho+ cells to petites within two to four generations after a shift of the culture from glucose to galactose medium. Overexpression of Abf2p in rho- cells also leads to a loss of mtDNA, but at a slower rate than was observed for rho+ cells. The mtDNA instability phenotype is related to the DNA-binding properties of Abf2p because a mutant Abf2p that contains mutations in residues of both HMG box domains known to affect DNA binding in vitro, and that binds poorly to mtDNA in vivo, complements deltaabf2 cells only weakly and greatly lessens the effect of overproduction on mtDNA instability. In vivo binding was assessed by colocalization to mtDNA of fusions between mutant or wild-type Abf2p and green fluorescent protein.These findings are discussed in the context of a model relating mtDNA copy number control and stability to mtDNA recombination.

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Selected References

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

  1. Asai T., Sommer S., Bailone A., Kogoma T. Homologous recombination-dependent initiation of DNA replication from DNA damage-inducible origins in Escherichia coli. EMBO J. 1993 Aug;12(8):3287–3295. doi: 10.1002/j.1460-2075.1993.tb05998.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Azpiroz R., Butow R. A. Patterns of mitochondrial sorting in yeast zygotes. Mol Biol Cell. 1993 Jan;4(1):21–36. doi: 10.1091/mbc.4.1.21. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Baldacci G., Chérif-Zahar B., Bernardi G. The initiation of DNA replication in the mitochondrial genome of yeast. EMBO J. 1984 Sep;3(9):2115–2120. doi: 10.1002/j.1460-2075.1984.tb02099.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Baxevanis A. D., Landsman D. The HMG-1 box protein family: classification and functional relationships. Nucleic Acids Res. 1995 May 11;23(9):1604–1613. doi: 10.1093/nar/23.9.1604. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Bendich A. J. Structural analysis of mitochondrial DNA molecules from fungi and plants using moving pictures and pulsed-field gel electrophoresis. J Mol Biol. 1996 Feb 2;255(4):564–588. doi: 10.1006/jmbi.1996.0048. [DOI] [PubMed] [Google Scholar]
  6. Berben G., Dumont J., Gilliquet V., Bolle P. A., Hilger F. The YDp plasmids: a uniform set of vectors bearing versatile gene disruption cassettes for Saccharomyces cerevisiae. Yeast. 1991 Jul;7(5):475–477. doi: 10.1002/yea.320070506. [DOI] [PubMed] [Google Scholar]
  7. Caron F., Jacq C., Rouvière-Yaniv J. Characterization of a histone-like protein extracted from yeast mitochondria. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4265–4269. doi: 10.1073/pnas.76.9.4265. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Chen D. C., Yang B. C., Kuo T. T. One-step transformation of yeast in stationary phase. Curr Genet. 1992 Jan;21(1):83–84. doi: 10.1007/BF00318659. [DOI] [PubMed] [Google Scholar]
  9. Dairaghi D. J., Shadel G. S., Clayton D. A. Addition of a 29 residue carboxyl-terminal tail converts a simple HMG box-containing protein into a transcriptional activator. J Mol Biol. 1995 May 26;249(1):11–28. doi: 10.1006/jmbi.1995.9889. [DOI] [PubMed] [Google Scholar]
  10. Diffley J. F., Stillman B. A close relative of the nuclear, chromosomal high-mobility group protein HMG1 in yeast mitochondria. Proc Natl Acad Sci U S A. 1991 Sep 1;88(17):7864–7868. doi: 10.1073/pnas.88.17.7864. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Diffley J. F., Stillman B. DNA binding properties of an HMG1-related protein from yeast mitochondria. J Biol Chem. 1992 Feb 15;267(5):3368–3374. [PubMed] [Google Scholar]
  12. Evans D. H., Kolodner R. Effect of DNA structure and nucleotide sequence on Holliday junction resolution by a Saccharomyces cerevisiae endonuclease. J Mol Biol. 1988 May 5;201(1):69–80. doi: 10.1016/0022-2836(88)90439-1. [DOI] [PubMed] [Google Scholar]
  13. Falciola L., Murchie A. I., Lilley D. M., Bianchi M. Mutational analysis of the DNA binding domain A of chromosomal protein HMG1. Nucleic Acids Res. 1994 Feb 11;22(3):285–292. doi: 10.1093/nar/22.3.285. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Fangman W. L., Henly J. W., Brewer B. J. RPO41-independent maintenance of [rho-] mitochondrial DNA in Saccharomyces cerevisiae. Mol Cell Biol. 1990 Jan;10(1):10–15. doi: 10.1128/mcb.10.1.10. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Fisher R. P., Clayton D. A. Purification and characterization of human mitochondrial transcription factor 1. Mol Cell Biol. 1988 Aug;8(8):3496–3509. doi: 10.1128/mcb.8.8.3496. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Fisher R. P., Lisowsky T., Parisi M. A., Clayton D. A. DNA wrapping and bending by a mitochondrial high mobility group-like transcriptional activator protein. J Biol Chem. 1992 Feb 15;267(5):3358–3367. [PubMed] [Google Scholar]
  17. Fisher R. P., Topper J. N., Clayton D. A. Promoter selection in human mitochondria involves binding of a transcription factor to orientation-independent upstream regulatory elements. Cell. 1987 Jul 17;50(2):247–258. doi: 10.1016/0092-8674(87)90220-0. [DOI] [PubMed] [Google Scholar]
  18. Formosa T., Alberts B. M. DNA synthesis dependent on genetic recombination: characterization of a reaction catalyzed by purified bacteriophage T4 proteins. Cell. 1986 Dec 5;47(5):793–806. doi: 10.1016/0092-8674(86)90522-2. [DOI] [PubMed] [Google Scholar]
  19. Greenleaf A. L., Kelly J. L., Lehman I. R. Yeast RPO41 gene product is required for transcription and maintenance of the mitochondrial genome. Proc Natl Acad Sci U S A. 1986 May;83(10):3391–3394. doi: 10.1073/pnas.83.10.3391. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Hall R. M., Nagley P., Linnane A. W. Biogenesis of mitochondria. XLII. Genetic analysis of the control of cellular mitochondrial DNA levels in Saccharomyces cerevisiae. Mol Gen Genet. 1976 May 7;145(2):169–175. doi: 10.1007/BF00269590. [DOI] [PubMed] [Google Scholar]
  21. Herskowitz I., Jensen R. E. Putting the HO gene to work: practical uses for mating-type switching. Methods Enzymol. 1991;194:132–146. doi: 10.1016/0076-6879(91)94011-z. [DOI] [PubMed] [Google Scholar]
  22. Hoffman C. S., Winston F. A ten-minute DNA preparation from yeast efficiently releases autonomous plasmids for transformation of Escherichia coli. Gene. 1987;57(2-3):267–272. doi: 10.1016/0378-1119(87)90131-4. [DOI] [PubMed] [Google Scholar]
  23. Kao L. R., Megraw T. L., Chae C. B. Essential role of the HMG domain in the function of yeast mitochondrial histone HM: functional complementation of HM by the nuclear nonhistone protein NHP6A. Proc Natl Acad Sci U S A. 1993 Jun 15;90(12):5598–5602. doi: 10.1073/pnas.90.12.5598. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Kleff S., Kemper B., Sternglanz R. Identification and characterization of yeast mutants and the gene for a cruciform cutting endonuclease. EMBO J. 1992 Feb;11(2):699–704. doi: 10.1002/j.1460-2075.1992.tb05102.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Lockshon D., Zweifel S. G., Freeman-Cook L. L., Lorimer H. E., Brewer B. J., Fangman W. L. A role for recombination junctions in the segregation of mitochondrial DNA in yeast. Cell. 1995 Jun 16;81(6):947–955. doi: 10.1016/0092-8674(95)90014-4. [DOI] [PubMed] [Google Scholar]
  26. Lopez I. C., Farrelly F., Butow R. A. Trans action and the var1 determinant region on yeast mitochondrial DNA. Specific labeling of mitochondrial translation products in zygotes. J Biol Chem. 1981 Jun 25;256(12):6496–6501. [PubMed] [Google Scholar]
  27. Lorimer H. E., Brewer B. J., Fangman W. L. A test of the transcription model for biased inheritance of yeast mitochondrial DNA. Mol Cell Biol. 1995 Sep;15(9):4803–4809. doi: 10.1128/mcb.15.9.4803. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Luder A., Mosig G. Two alternative mechanisms for initiation of DNA replication forks in bacteriophage T4: priming by RNA polymerase and by recombination. Proc Natl Acad Sci U S A. 1982 Feb;79(4):1101–1105. doi: 10.1073/pnas.79.4.1101. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Maleszka R., Skelly P. J., Clark-Walker G. D. Rolling circle replication of DNA in yeast mitochondria. EMBO J. 1991 Dec;10(12):3923–3929. doi: 10.1002/j.1460-2075.1991.tb04962.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Megraw T. L., Chae C. B. Functional complementarity between the HMG1-like yeast mitochondrial histone HM and the bacterial histone-like protein HU. J Biol Chem. 1993 Jun 15;268(17):12758–12763. [PubMed] [Google Scholar]
  31. Myers A. M., Tzagoloff A., Kinney D. M., Lusty C. J. Yeast shuttle and integrative vectors with multiple cloning sites suitable for construction of lacZ fusions. Gene. 1986;45(3):299–310. doi: 10.1016/0378-1119(86)90028-4. [DOI] [PubMed] [Google Scholar]
  32. Nagley P., Linnane A. W. Biogenesis of mitochondria. XXI. Studies on the nature of the mitochondrial genome in yeast: the degenerative effects of ethidium bromide on mitochondrial genetic information in a respiratory competent strain. J Mol Biol. 1972 Apr 28;66(1):181–193. doi: 10.1016/s0022-2836(72)80015-9. [DOI] [PubMed] [Google Scholar]
  33. Newman S. M., Zelenaya-Troitskaya O., Perlman P. S., Butow R. A. Analysis of mitochondrial DNA nucleoids in wild-type and a mutant strain of Saccharomyces cerevisiae that lacks the mitochondrial HMG box protein Abf2p. Nucleic Acids Res. 1996 Jan 15;24(2):386–393. doi: 10.1093/nar/24.2.386. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Ng R., Abelson J. Isolation and sequence of the gene for actin in Saccharomyces cerevisiae. Proc Natl Acad Sci U S A. 1980 Jul;77(7):3912–3916. doi: 10.1073/pnas.77.7.3912. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Nunnari J., Marshall W. F., Straight A., Murray A., Sedat J. W., Walter P. Mitochondrial transmission during mating in Saccharomyces cerevisiae is determined by mitochondrial fusion and fission and the intramitochondrial segregation of mitochondrial DNA. Mol Biol Cell. 1997 Jul;8(7):1233–1242. doi: 10.1091/mbc.8.7.1233. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. OGUR M., ST. JOHN R., NAGAI S. Tetrazolium overlay technique for population studies of respiration deficiency in yeast. Science. 1957 May 10;125(3254):928–929. doi: 10.1126/science.125.3254.928. [DOI] [PubMed] [Google Scholar]
  37. Parikh V. S., Conrad-Webb H., Docherty R., Butow R. A. Interaction between the yeast mitochondrial and nuclear genomes influences the abundance of novel transcripts derived from the spacer region of the nuclear ribosomal DNA repeat. Mol Cell Biol. 1989 May;9(5):1897–1907. doi: 10.1128/mcb.9.5.1897. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Parisi M. A., Clayton D. A. Similarity of human mitochondrial transcription factor 1 to high mobility group proteins. Science. 1991 May 17;252(5008):965–969. doi: 10.1126/science.2035027. [DOI] [PubMed] [Google Scholar]
  39. Parisi M. A., Xu B., Clayton D. A. A human mitochondrial transcriptional activator can functionally replace a yeast mitochondrial HMG-box protein both in vivo and in vitro. Mol Cell Biol. 1993 Mar;13(3):1951–1961. doi: 10.1128/mcb.13.3.1951. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Preiser P. R., Wilson R. J., Moore P. W., McCready S., Hajibagheri M. A., Blight K. J., Strath M., Williamson D. H. Recombination associated with replication of malarial mitochondrial DNA. EMBO J. 1996 Feb 1;15(3):684–693. [PMC free article] [PubMed] [Google Scholar]
  41. Strausberg R. L., Perlman P. S. The effect of zygotic bud position on the transmission of mitochondrial genes in Saccharomyces cerevisiae. Mol Gen Genet. 1978 Jul 11;163(2):131–144. doi: 10.1007/BF00267404. [DOI] [PubMed] [Google Scholar]
  42. Teo S. H., Grasser K. D., Hardman C. H., Broadhurst R. W., Laue E. D., Thomas J. O. Two mutations in the HMG-box with very different structural consequences provide insights into the nature of binding to four-way junction DNA. EMBO J. 1995 Aug 1;14(15):3844–3853. doi: 10.1002/j.1460-2075.1995.tb00054.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Tzagoloff A., Myers A. M. Genetics of mitochondrial biogenesis. Annu Rev Biochem. 1986;55:249–285. doi: 10.1146/annurev.bi.55.070186.001341. [DOI] [PubMed] [Google Scholar]
  44. Vida T. A., Graham T. R., Emr S. D. In vitro reconstitution of intercompartmental protein transport to the yeast vacuole. J Cell Biol. 1990 Dec;111(6 Pt 2):2871–2884. doi: 10.1083/jcb.111.6.2871. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Weislogel P. O., Butow R. A. Low temperature and chloramphenicol induction of respiratory deficiency in a cold-sensitive mutant of Saccharomyces cerevisiae. Proc Natl Acad Sci U S A. 1970 Sep;67(1):52–58. doi: 10.1073/pnas.67.1.52. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Williamson D. H., Fennell D. J. Visualization of yeast mitochondrial DNA with the fluorescent stain "DAPI". Methods Enzymol. 1979;56:728–733. doi: 10.1016/0076-6879(79)56065-0. [DOI] [PubMed] [Google Scholar]
  47. Zinn A. R., Pohlman J. K., Perlman P. S., Butow R. A. Kinetic and segregational analysis of mitochondrial DNA recombination in yeast. Plasmid. 1987 May;17(3):248–256. doi: 10.1016/0147-619x(87)90033-3. [DOI] [PubMed] [Google Scholar]
  48. Zweifel S. G., Fangman W. L. A nuclear mutation reversing a biased transmission of yeast mitochondrial DNA. Genetics. 1991 Jun;128(2):241–249. doi: 10.1093/genetics/128.2.241. [DOI] [PMC free article] [PubMed] [Google Scholar]
  49. van Gent D. C., Hiom K., Paull T. T., Gellert M. Stimulation of V(D)J cleavage by high mobility group proteins. EMBO J. 1997 May 15;16(10):2665–2670. doi: 10.1093/emboj/16.10.2665. [DOI] [PMC free article] [PubMed] [Google Scholar]

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