Abstract
The relationship between the chloramphenicol (CAP)-resistant phenotype and the mtDNA genotype was investigated in segregating human, HeLa × HT1080, somatic cell hybrids. The parental mtDNAs were quantitated in heteroplasmic cells by using restriction fragment length polymorphisms (RFLPs) detected in Southern blots. CAP-resistant (R) × CAP-sensitive (S) hybrids selected and grown in CAP for brief periods had as little as 25% CAP-R mtDNA. With prolonged selection, the CAP-R mtDNA increased to 90–95%. Hybrids selected and passaged without CAP either retained both mtDNAs or progressively lost one mtDNA (mitotic segregation). The CAP-resistance phenotype of these hybrids changed abruptly when the proportion of CAP-R mtDNAs fluctuated around approximately 10% (threshold effect). Hybrids with greater than 25% HT1080 mtDNA had an additional characteristic. They cloned better with CAP than without. The cloning efficiency in CAP of hybrids having 90% HT1080 mtDNA was more than fivefold greater than the control.
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Bogenhagen, D., and Clayton, D.A. (1974).J. Biol. Chem. 249:7991–7995.
Shmooker Reis, R.J., and Goldstein, S. (1983).J. Biol. Chem. 258:9078–9085.
Anderson, S., Banker, A.T., Barrell, B.G., de Bruijin, M.H.L., Coulson, A.R., Drouin, J., Eperon, I.C., Nierlich, D.P., Rose, B.A., Sanger, F., Schreier, P.H., Smith, A.J.H., Staden, R., and Young, I.G. (1981).Nature 296:457–465.
Chomyn, A., Mariottini, P., Cleeter, M.W.J., Ragan, C.I., Matsuno-Yagi, A., Hatefi, Y., Doolittle, R.F., and Attardi, G. (1985).Nature 314:592–597.
Wallace, D.C., Yang, J., Ye, J., Lott, M.T., Oliver, N.A., and McCarthy, J. (1986).Am. J. Hum. Genet. (in press).
Bunn, C.L., Wallace, D.C., and Eisenstadt, J.M. (1977).Somat. Cell Genet. 3:71–92.
Wallace, D.C., Bunn, C.L., and Eisenstadt, J.M. (1977).Somat. Cell Genet. 3:93–119.
Wallace, D.C. (1982). InTechniques in Somatic Cell Genetics, (ed.) Shay, J.W. (Plenum Publishing, New York), pp. 159–187.
Howell, N. (1983).Somat. Cell Genet. 9:1–24.
Ho, C., and Coon, H.G. (1979). InExtrachromosomal DNA ICN-UCLA Symposium on Molecular and Cellular Biology, (eds.) Cummings, D.J., Borst, P., Dawid, I.B., Weissman, S.M., and Fox, C.F. (Academic Press, New York), pp. 501–514.
Wallace, D.C. (1981).Mol. Cell Biol. 1:697–710.
Howell, N., Huang, P., and Kolodner, R.D. (1984).Somat. Cell. Mol. Genet. 10:259–274.
Wallace, D.C., Bunn, C.L., and Eisenstadt, J.M. (1975).J. Cell Biol. 67:174–188.
Shipman, C., Jr., Smith, S.H., and Droch, J.C. (1972).Proc. Natl. Acad. Sci. U.S.A. 69:1735–1757.
Wahl, G.M., Stern, M., and Stark, G.R. (1979).Proc. Nail. Acad. Sci. U.S.A. 76:3683–3687.
Maniatis, T., Fritsch, E.F., and Sambrook, J. (1982).Molecular Cloning: A Laboratory Manual. (Cold Spring Harbor Laboratory, Cold Spring Harbor, New York), p. 545.
Giles, R.E., Stroynowski, I., and Wallace, D.C. (1980).Somat. Cell Genet. 6:543–554.
Oliver, N.A., and Wallace, D.C. (1982).Mol. Cell Biol. 2:30–41.
White, F.A., and Bunn, C.L. (1984). Mol. Gen. Genet.197:453–460.
Kearsey, S.E., Munro, E., and Craig, I.W. (1985). Proc. R. Soc. Lond.B224:315–323.
Attardi, G., Cantatore, P., Chomyn, A., Crews, S., Gelfand, R., Merkel, C., Montoya, J., & Ojala, D. (1982). InMitochondrial Genes, (eds.) Slonimski, P., Borst, P., and Attardi, G., (Cold Spring Harbor Laboratory, Cold Spring Harbor, New York), pp. 51–71.
Ziegler, M.L., and Davidson, R.L. (1981).Somat. Cell Genet. 7:73–88.
Oliver, N.A., Greenberg, B.D., and Wallace, D.C. (1983).J. Biol. Chem. 258:5834–5839.
Yatscoff, R.W., Goldstein, S., and Freeman, K.B. (1978).Somat. Cell Genet. 4:633–645.
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Wallace, D.C. Mitotic segregation of mitochondrial dnas in human cell hybrids and expression of chloramphenicol resistance. Somat Cell Mol Genet 12, 41–49 (1986). https://doi.org/10.1007/BF01560726
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DOI: https://doi.org/10.1007/BF01560726