Skip to main content
PMC home page
Molecular and Cellular Biology logoLink to Molecular and Cellular Biology
. 1986 Dec;6(12):4509–4515. doi: 10.1128/mcb.6.12.4509

Mitochondrial and nonmitochondrial citrate synthases in Saccharomyces cerevisiae are encoded by distinct homologous genes.

M Rosenkrantz, T Alam, K S Kim, B J Clark, P A Srere, L P Guarente
PMCID: PMC367235  PMID: 3540614

Abstract

Saccharomyces cerevisiae contains two genes, CIT1 and CIT2, encoding functional citrate synthase (K.-S. Kim, M. S. Rosenkrantz, and L. Guarente, Mol. Cell. Biol. 6:1936-1942, 1986). We show here that CIT2 encodes a nonmitochondrial form of citrate synthase. The DNA sequence of CIT2 presented provides a possible explanation for why the CIT2 product, unlike the CIT1 product, fails to be imported into mitochondria. While the products of these two genes are highly homologous, they diverge strikingly at their amino termini. The amino terminus of the CIT1 primary translation product extends 39 residues beyond the amino termini of Escherichia coli and porcine citrate synthases. This extension consists of a typical mitochondrial targeting motif. The amino terminus of the CIT2 primary translation product extends 20 residues beyond the amino termini of the E. coli and porcine enzymes. The CIT2-encoded extension is not homologous to that of CIT1, resulting in a nonmitochondrial localization of the product. The CIT2-encoded extension, however, does bear certain similarities to mitochondrial targeting sequences. The possible role of this sequence in targeting this CIT2 product to a nonmitochondrial organelle is discussed.

Full text

PDF
4509

Selected References

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

  1. Alam T., Finkelstein D., Srere P. A. In vitro translation of mRNA for yeast citrate synthase. J Biol Chem. 1982 Sep 25;257(18):11181–11185. [PubMed] [Google Scholar]
  2. Avers C. J., Federman M. The occurrence in yeast of cytoplasmic granules which resemble microbodies. J Cell Biol. 1968 May;37(2):555–559. doi: 10.1083/jcb.37.2.555. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Beltzer J. P., Chang L. F., Hinkkanen A. E., Kohlhaw G. B. Structure of yeast LEU4. The 5' flanking region contains features that predict two modes of control and two productive translation starts. J Biol Chem. 1986 Apr 15;261(11):5160–5167. [PubMed] [Google Scholar]
  4. Bennetzen J. L., Hall B. D. Codon selection in yeast. J Biol Chem. 1982 Mar 25;257(6):3026–3031. [PubMed] [Google Scholar]
  5. Bloxham D. P., Parmelee D. C., Kumar S., Wade R. D., Ericsson L. H., Neurath H., Walsh K. A., Titani K. Primary structure of porcine heart citrate synthase. Proc Natl Acad Sci U S A. 1981 Sep;78(9):5381–5385. doi: 10.1073/pnas.78.9.5381. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Böhni P. C., Daum G., Schatz G. Import of proteins into mitochondria. Partial purification of a matrix-located protease involved in cleavage of mitochondrial precursor polypeptides. J Biol Chem. 1983 Apr 25;258(8):4937–4943. [PubMed] [Google Scholar]
  7. Cooper T. G., Beevers H. Mitochondria and glyoxysomes from castor bean endosperm. Enzyme constitutents and catalytic capacity. J Biol Chem. 1969 Jul 10;244(13):3507–3513. [PubMed] [Google Scholar]
  8. Daum G., Böhni P. C., Schatz G. Import of proteins into mitochondria. Cytochrome b2 and cytochrome c peroxidase are located in the intermembrane space of yeast mitochondria. J Biol Chem. 1982 Nov 10;257(21):13028–13033. [PubMed] [Google Scholar]
  9. Douglas M. G., McCammon M. T., Vassarotti A. Targeting proteins into mitochondria. Microbiol Rev. 1986 Jun;50(2):166–178. doi: 10.1128/mr.50.2.166-178.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Duntze W., Neumann D., Gancedo J. M., Atzpodien W., Holzer H. Studies on the regulation and localization of the glyoxylate cycle enzymes in Saccharomyces cerevisiae. Eur J Biochem. 1969 Aug;10(1):83–89. doi: 10.1111/j.1432-1033.1969.tb00658.x. [DOI] [PubMed] [Google Scholar]
  11. Holland M. J., Holland J. P., Thill G. P., Jackson K. A. The primary structures of two yeast enolase genes. Homology between the 5' noncoding flanking regions of yeast enolase and glyceraldehyde-3-phosphate dehydrogenase genes. J Biol Chem. 1981 Feb 10;256(3):1385–1395. [PubMed] [Google Scholar]
  12. Hoosein M. A., Lewin A. S. Derepression of citrate synthase in Saccharomyces cerevisiae may occur at the level of transcription. Mol Cell Biol. 1984 Feb;4(2):247–253. doi: 10.1128/mcb.4.2.247. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Hopper A. K., Furukawa A. H., Pham H. D., Martin N. C. Defects in modification of cytoplasmic and mitochondrial transfer RNAs are caused by single nuclear mutations. Cell. 1982 Mar;28(3):543–550. doi: 10.1016/0092-8674(82)90209-4. [DOI] [PubMed] [Google Scholar]
  14. Kim K. S., Rosenkrantz M. S., Guarente L. Saccharomyces cerevisiae contains two functional citrate synthase genes. Mol Cell Biol. 1986 Jun;6(6):1936–1942. doi: 10.1128/mcb.6.6.1936. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  16. Laz T. M., Pietras D. F., Sherman F. Differential regulation of the duplicated isocytochrome c genes in yeast. Proc Natl Acad Sci U S A. 1984 Jul;81(14):4475–4479. doi: 10.1073/pnas.81.14.4475. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Linn T. C. Studies on the apparent instability of bovine kidney alpha-ketoglutarate dehydrogenase complex. Arch Biochem Biophys. 1974 Apr 2;161(2):505–514. doi: 10.1016/0003-9861(74)90333-6. [DOI] [PubMed] [Google Scholar]
  18. Martin N. C., Hopper A. K. Isopentenylation of both cytoplasmic and mitochondrial tRNA is affected by a single nuclear mutation. J Biol Chem. 1982 Sep 25;257(18):10562–10565. [PubMed] [Google Scholar]
  19. McAlister L., Holland M. J. Targeted deletion of a yeast enolase structural gene. Identification and isolation of yeast enolase isozymes. J Biol Chem. 1982 Jun 25;257(12):7181–7188. [PubMed] [Google Scholar]
  20. Natsoulis G., Hilger F., Fink G. R. The HTS1 gene encodes both the cytoplasmic and mitochondrial histidine tRNA synthetases of S. cerevisiae. Cell. 1986 Jul 18;46(2):235–243. doi: 10.1016/0092-8674(86)90740-3. [DOI] [PubMed] [Google Scholar]
  21. Perlman P. S., Mahler H. R. Intracellular localization of enzymes in yeast. Arch Biochem Biophys. 1970 Jan;136(1):245–259. doi: 10.1016/0003-9861(70)90348-6. [DOI] [PubMed] [Google Scholar]
  22. Remington S., Wiegand G., Huber R. Crystallographic refinement and atomic models of two different forms of citrate synthase at 2.7 and 1.7 A resolution. J Mol Biol. 1982 Jun 15;158(1):111–152. doi: 10.1016/0022-2836(82)90452-1. [DOI] [PubMed] [Google Scholar]
  23. Rickey T. M., Lewin A. S. Extramitochondrial citrate synthase activity in bakers' yeast. Mol Cell Biol. 1986 Feb;6(2):488–493. doi: 10.1128/mcb.6.2.488. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Struhl K. The yeast his3 promoter contains at least two distinct elements. Proc Natl Acad Sci U S A. 1982 Dec;79(23):7385–7389. doi: 10.1073/pnas.79.23.7385. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Suissa M., Suda K., Schatz G. Isolation of the nuclear yeast genes for citrate synthase and fifteen other mitochondrial proteins by a new screening method. EMBO J. 1984 Aug;3(8):1773–1781. doi: 10.1002/j.1460-2075.1984.tb02045.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Young E. T., Pilgrim D. Isolation and DNA sequence of ADH3, a nuclear gene encoding the mitochondrial isozyme of alcohol dehydrogenase in Saccharomyces cerevisiae. Mol Cell Biol. 1985 Nov;5(11):3024–3034. doi: 10.1128/mcb.5.11.3024. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. van Loon A. P., Young E. T. Intracellular sorting of alcohol dehydrogenase isoenzymes in yeast: a cytosolic location reflects absence of an amino-terminal targeting sequence for the mitochondrion. EMBO J. 1986 Jan;5(1):161–165. doi: 10.1002/j.1460-2075.1986.tb04191.x. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Molecular and Cellular Biology are provided here courtesy of Taylor & Francis

RESOURCES