Summary
This study details the characteristics of two temperature-conditional pet mutants of yeast, strains ts1860 and ts379, which at the non-permissive temperature show deficiencies in the formation of three mitochondrially encoded subunits of the ATP synthase complex. By analysis of mitochondrial translation products, and of mitochondrial transcription in temperature shift experiments from the permissive (22°C) to the non-permissive (36°C) temperature, it was concluded that the nuclear mutations in both mutants primarily inhibit synthesis of ATP synthase subunit 9, and that reductions in subunit 8 and 6 synthesis are secondary pleiotropic effects. Following transfer to 36°C, cells of mutant ts379 display a near complete inhibition of subunit 9 synthesis within 1 h, coincident with a marked reduction in the level of the cognate oli1 mRNA. On the other hand, near complete inhibition of subunit 9 synthesis in strain ts1860 occurs after 3 h at 36°C, at which time there is little change in the level of subunit 9 mRNA. In both mutants the mRNA levels for subunits 6 and 8 are not significantly affected at the time of inhibition of subunit 9 synthesis. Provision of an alternative source of subunit 8, translated extra-mitochondrially for import into the organelle, does not overcome the mutant phenotype of either mutant at 36°C, confirming that subunit 8 is not the sole or primary deficiency in each mutant. The mutants indicate that the products of a least two nuclear genes (designated AEP1 and AEP2) are required for the expression of the mitochondrial oli1 gene and the synthesis of subunit 9. The product of the AEP1 gene (defective in mutant ts1860) is required for translation of oli1 mRNA while the AEP2 product (defective in mutant ts379) is essential either for the stability of oli1 mRNA or for the correct processing of precursor transcripts to the mature message.
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References
Ackerman SH, Tzagoloff A (1990) J Biol Chem 265:9952–9959
Bingham CG, Nagley P (1983) Biochim Biophys Acta 740:88–98
Boy-Marcotte E, Jacquet M (1982) Gene 20:433–440
Burkl G, Demmer W, Holzner H, Schweizer E (1976) In: Bandlow W, Schweyen RJ, Thomas DY, Wolf K, Kaudewitz F (eds) Genetics, biogenesis and bioenergetics of mitochondria. W de Gruyter, Berlin, pp 39–48
Cobon GS, Beilharz MW, Linnane AW, Nagley P (1982) Curr Genet 5:97–107
Costanzo MC, Fox TD (1988) Proc Natl Acad Sci USA 85:2677–2681
Decoster E, Simon M, Hatat D, Faye G (1990) Mol Gen Genet 224:111–118
Dieckmann CL, Tzagoloff A (1985) J Biol Chem 260:1513–1520
Dieckmann CL, Mittelmeier TM (1987) Curr Genet 12:391–397
Dieckmann CL, Bonitz SG, Hill J, Homison G, McGraw P, Pape L, Thalenfeld BE, Tzagoloff A (1982) In: Slonimski PP, Borst P, Attardi G (eds) Mitochondrial genes. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York, pp 213–223
Dieckmann CL, Koerner TJ, Tzagoloff (1984) J Biol Chem 259:4722–4731
Ebner E, Mennucci L, Schatz G (1973a) J Biol Chem 248:5360–5368
Ebner E, Mason TL, Schatz G (1973b) J Biol Chem 248:5369–5378
Fox TD (1986) Trends Genet Sci 2:97–100
Grivell LA (1989) Eur J Biochem 182:477–493
Hadikusumo RG, Meltzer S, Choo WM, Jean-Francois MJB, Linnane AW, Marzuki S (1988) Biochim Biophys Acta 933:212–222
Jean-Francois MJB, Hadikusumo RG, Watkins LC, Lukins HB, Linuane AW, Marzuki S (1986a) Biochim Biophys Acta 852:133–143
Jean-Francois MJB, Lukins HB, Marzuki S (1986b) Biochim Biophys Acta 868:178–182
Kloeckener-Gruissem B, McEwen JE, Poyton RO (1987) Curr Genet 12:311–322
Lang B, Gurger G, Doxiadis I, Thomas DY, Bandlow W, Kaudewitz F (1977) Anal Biochem 77:110–121
McEwen JE, Ko C, Kloeckener-Gruissem B, Poyton RO (1986) J Biol Chem 261:11872–11879
Macino G, Tzagoloff A (1980) Cell 20:507–517
Macreadie IG, Novitski CE, Maxwell RJ, John U, Ooi BG, McMullen GL, Lukins HB, Linnane AW, Nagley P (1983) Nucleic Acids Res 11:4435–4451
Michaelis G, Mannhaupt G, Pratje E, Fischer E, Naggert J, Schweizer E (1982) In: Slonimski PP, Borst P, Attardi G (eds) Mitochondrial genes. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York, pp 311–321
Muller PP, Reif MK, Zonghou S, Sengstag C, Mason TL, Fox TD (1984) J Molec Biol 175:431–452
Murphy M, Choo KB, Macreadie I, Marzuki S, Lukins HB, Nagley P, Linnane AW (1980) Arch Biochem Biophys 203:260–270
Nagley P, Farrell LB, Gearing DP, Nero D, Meltzer S, Devenish RJ (1988) Proc Natl Acad Sci USA 85:2091–2095
Nagley P (1988) Trends Genet Sci 4:46–52
Ooi BG, Nagley P (1986) Curr Genet 10:713–723
Ooi BG, Lukins HB, Linnane AW, Nagley P (1987) Nucleic Acids Res 15:1965–1977
Pillar T, Lang BF, Steinberger I, Vogt B, Kaudewitz F (1983) J Biol Chem 258:7954–7959
Poutre CG, Fox TD (1987) Genetics 115:637–647
Pratje E, Guiard B (1986) EMBO J 5:1313–1317
Rodel G (1986) Curr Genet 11:41–45
Rodel G, Fox TD (1987) Mol Gen Genet 206:45–50
Rodel G, Michaelis U, Forsbach V, Kreike J, Kaudewitz F (1986) Curr Genet 11:47–53
Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York
Schulze M, Rodel G (1988) Mol Gen Genet 211:492–498
Schweizer E, Demmer W, Holzner U, Tahedl HW (1977) In: Bandlow W, Schweyen RJ, Wolf K, Kaudewitz F (eds) Mitochondria 1977. Genetics and biogenesis of mitochondria. Walter de Gruyter, Berlin, pp 91–105
Sherman F, Fink GR, Hicks JP (1986) Laboratory course manual for methods in yeast genetics. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York
Simon M, Faye G (1984) Mol Gen Genet 196:266–274
Smooker PM (1987) PhD thesis, Monash University
Studier FW (1973) J Molec Biol 79:237–248
Tzagoloff A, Dieckmann CL (1990) Microb Rev 54:211–225
Tzagoloff A, Myers AM (1986) Annu Rev Biochem 55:249–285
Zassenhaus HP, Martin NC, Butow RA (1984) J Biol Chem 259:6019–6027
Zennaro E, Grimaldi L, Baldacci G, Frontali L (1985) Eur J Biochem 147:191–196
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Communicated by R. J. Schweyen
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Payne, M.J., Schweizer, E. & Lukins, H.B. Properties of two nuclear pet mutants affecting expression of the mitochondrial oli1 gene of Saccharomyces cerevisiae . Curr Genet 19, 343–351 (1991). https://doi.org/10.1007/BF00309594
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DOI: https://doi.org/10.1007/BF00309594