Skip to main content
PMC home page
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1996 Aug 2;134(4):949–961. doi: 10.1083/jcb.134.4.949

Aberrant mitosis in fission yeast mutants defective in fatty acid synthetase and acetyl CoA carboxylase

PMCID: PMC2120970  PMID: 8769419

Abstract

Two fission yeast temperature-sensitive mutants, cut6 and lsd1, show a defect in nuclear division. The daughter nuclei differ dramatically in size (the phenotype designated lsd, large and small daughter). Fluorescence in situ hybridization (FISH) revealed that sister chromatids were separated in the lsd cells, but appeared highly compact in one of the two daughter nuclei. EM showed asymmetric nuclear elongation followed by unequal separation of nonchromosomal nuclear structures in these mutant nuclei. The small nuclei lacked electron- dense nuclear materials and contained highly compacted chromatin. The cut6+ and lsd1+ genes are essential for viability and encode, respectively, acetyl CoA carboxylase and fatty acid synthetase, the key enzymes for fatty acid synthesis. Gene disruption of lsd1+ led to the lsd phenotype. Palmitate in medium fully suppressed the phenotypes of lsd1. Cerulenin, an inhibitor for fatty acid synthesis, produced the lsd phenotype in wild type. The drug caused cell inviability during mitosis but not during the G2-arrest induced by the cdc25 mutation. A reduced level of fatty acid thus led to impaired separation of non- chromosomal nuclear components. We propose that fatty acid is directly or indirectly required for separating the mother nucleus into two equal daughters.

Full Text

The Full Text of this article is available as a PDF (4.0 MB).

Selected References

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

  1. Adachi Y., Yanagida M. Higher order chromosome structure is affected by cold-sensitive mutations in a Schizosaccharomyces pombe gene crm1+ which encodes a 115-kD protein preferentially localized in the nucleus and its periphery. J Cell Biol. 1989 Apr;108(4):1195–1207. doi: 10.1083/jcb.108.4.1195. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Al-Feel W., Chirala S. S., Wakil S. J. Cloning of the yeast FAS3 gene and primary structure of yeast acetyl-CoA carboxylase. Proc Natl Acad Sci U S A. 1992 May 15;89(10):4534–4538. doi: 10.1073/pnas.89.10.4534. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Anderson J. T., Paddy M. R., Swanson M. S. PUB1 is a major nuclear and cytoplasmic polyadenylated RNA-binding protein in Saccharomyces cerevisiae. Mol Cell Biol. 1993 Oct;13(10):6102–6113. doi: 10.1128/mcb.13.10.6102. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Casey P. J. Lipid modifications of G proteins. Curr Opin Cell Biol. 1994 Apr;6(2):219–225. doi: 10.1016/0955-0674(94)90139-2. [DOI] [PubMed] [Google Scholar]
  5. Chirala S. S. Coordinated regulation and inositol-mediated and fatty acid-mediated repression of fatty acid synthase genes in Saccharomyces cerevisiae. Proc Natl Acad Sci U S A. 1992 Nov 1;89(21):10232–10236. doi: 10.1073/pnas.89.21.10232. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Demeter J., Morphew M., Sazer S. A mutation in the RCC1-related protein pim1 results in nuclear envelope fragmentation in fission yeast. Proc Natl Acad Sci U S A. 1995 Feb 28;92(5):1436–1440. doi: 10.1073/pnas.92.5.1436. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Dietlein G., Schweizer E. Control of fatty-acid synthetase biosynthesis in Saccharomyces cerevisiae. Eur J Biochem. 1975 Oct 1;58(1):177–184. doi: 10.1111/j.1432-1033.1975.tb02362.x. [DOI] [PubMed] [Google Scholar]
  8. Ekwall K., Kermorgant M., Dujardin G., Groudinsky O., Slonimski P. P. The NAM8 gene in Saccharomyces cerevisiae encodes a protein with putative RNA binding motifs and acts as a suppressor of mitochondrial splicing deficiencies when overexpressed. Mol Gen Genet. 1992 May;233(1-2):136–144. doi: 10.1007/BF00587571. [DOI] [PubMed] [Google Scholar]
  9. Fankhauser C., Marks J., Reymond A., Simanis V. The S. pombe cdc16 gene is required both for maintenance of p34cdc2 kinase activity and regulation of septum formation: a link between mitosis and cytokinesis? EMBO J. 1993 Jul;12(7):2697–2704. doi: 10.1002/j.1460-2075.1993.tb05931.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Fukada Y., Takao T., Ohguro H., Yoshizawa T., Akino T., Shimonishi Y. Farnesylated gamma-subunit of photoreceptor G protein indispensable for GTP-binding. Nature. 1990 Aug 16;346(6285):658–660. doi: 10.1038/346658a0. [DOI] [PubMed] [Google Scholar]
  11. Funabashi H., Kawaguchi A., Tomoda H., Omura S., Okuda S., Iwasaki S. Binding site of cerulenin in fatty acid synthetase. J Biochem. 1989 May;105(5):751–755. doi: 10.1093/oxfordjournals.jbchem.a122739. [DOI] [PubMed] [Google Scholar]
  12. Funabiki H., Hagan I., Uzawa S., Yanagida M. Cell cycle-dependent specific positioning and clustering of centromeres and telomeres in fission yeast. J Cell Biol. 1993 Jun;121(5):961–976. doi: 10.1083/jcb.121.5.961. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Georgatos S. D., Meier J., Simos G. Lamins and lamin-associated proteins. Curr Opin Cell Biol. 1994 Jun;6(3):347–353. doi: 10.1016/0955-0674(94)90025-6. [DOI] [PubMed] [Google Scholar]
  14. Ha J., Daniel S., Broyles S. S., Kim K. H. Critical phosphorylation sites for acetyl-CoA carboxylase activity. J Biol Chem. 1994 Sep 2;269(35):22162–22168. [PubMed] [Google Scholar]
  15. Hagan I. M., Hyams J. S. The use of cell division cycle mutants to investigate the control of microtubule distribution in the fission yeast Schizosaccharomyces pombe. J Cell Sci. 1988 Mar;89(Pt 3):343–357. doi: 10.1242/jcs.89.3.343. [DOI] [PubMed] [Google Scholar]
  16. Hardie D. G., MacKintosh R. W. AMP-activated protein kinase--an archetypal protein kinase cascade? Bioessays. 1992 Oct;14(10):699–704. doi: 10.1002/bies.950141011. [DOI] [PubMed] [Google Scholar]
  17. Hasslacher M., Ivessa A. S., Paltauf F., Kohlwein S. D. Acetyl-CoA carboxylase from yeast is an essential enzyme and is regulated by factors that control phospholipid metabolism. J Biol Chem. 1993 May 25;268(15):10946–10952. [PubMed] [Google Scholar]
  18. Hirano T., Funahashi S., Uemura T., Yanagida M. Isolation and characterization of Schizosaccharomyces pombe cutmutants that block nuclear division but not cytokinesis. EMBO J. 1986 Nov;5(11):2973–2979. doi: 10.1002/j.1460-2075.1986.tb04594.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Holm C. Coming undone: how to untangle a chromosome. Cell. 1994 Jul 1;77(7):955–957. doi: 10.1016/0092-8674(94)90433-2. [DOI] [PubMed] [Google Scholar]
  20. Inokoshi J., Tomoda H., Hashimoto H., Watanabe A., Takeshima H., Omura S. Cerulenin-resistant mutants of Saccharomyces cerevisiae with an altered fatty acid synthase gene. Mol Gen Genet. 1994 Jul 8;244(1):90–96. doi: 10.1007/BF00280191. [DOI] [PubMed] [Google Scholar]
  21. Ito H., Fukuda Y., Murata K., Kimura A. Transformation of intact yeast cells treated with alkali cations. J Bacteriol. 1983 Jan;153(1):163–168. doi: 10.1128/jb.153.1.163-168.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Knobling A., Schweizer E. Temperature-sensitive mutants of the yeast fatty-acid-synthetase complex. Eur J Biochem. 1975 Nov 15;59(2):415–421. doi: 10.1111/j.1432-1033.1975.tb02469.x. [DOI] [PubMed] [Google Scholar]
  23. Koegl M., Zlatkine P., Ley S. C., Courtneidge S. A., Magee A. I. Palmitoylation of multiple Src-family kinases at a homologous N-terminal motif. Biochem J. 1994 Nov 1;303(Pt 3):749–753. doi: 10.1042/bj3030749. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Kokame K., Fukada Y., Yoshizawa T., Takao T., Shimonishi Y. Lipid modification at the N terminus of photoreceptor G-protein alpha-subunit. Nature. 1992 Oct 22;359(6397):749–752. doi: 10.1038/359749a0. [DOI] [PubMed] [Google Scholar]
  25. Koshland D. Mitosis: back to the basics. Cell. 1994 Jul 1;77(7):951–954. doi: 10.1016/0092-8674(94)90432-4. [DOI] [PubMed] [Google Scholar]
  26. Kuhajda F. P., Jenner K., Wood F. D., Hennigar R. A., Jacobs L. B., Dick J. D., Pasternack G. R. Fatty acid synthesis: a potential selective target for antineoplastic therapy. Proc Natl Acad Sci U S A. 1994 Jul 5;91(14):6379–6383. doi: 10.1073/pnas.91.14.6379. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Kuziora M. A., Chalmers J. H., Jr, Douglas M. G., Hitzeman R. A., Mattick J. S., Wakil S. J. Molecular cloning of fatty acid synthetase genes from Saccharomyces cerevisiae. J Biol Chem. 1983 Oct 10;258(19):11648–11653. [PubMed] [Google Scholar]
  28. Lee F. J., Moss J. An RNA-binding protein gene (RBP1) of Saccharomyces cerevisiae encodes a putative glucose-repressible protein containing two RNA recognition motifs. J Biol Chem. 1993 Jul 15;268(20):15080–15087. [PubMed] [Google Scholar]
  29. López-Casillas F., Bai D. H., Luo X. C., Kong I. S., Hermodson M. A., Kim K. H. Structure of the coding sequence and primary amino acid sequence of acetyl-coenzyme A carboxylase. Proc Natl Acad Sci U S A. 1988 Aug;85(16):5784–5788. doi: 10.1073/pnas.85.16.5784. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Matsumoto T., Beach D. Interaction of the pim1/spi1 mitotic checkpoint with a protein phosphatase. Mol Biol Cell. 1993 Mar;4(3):337–345. doi: 10.1091/mbc.4.3.337. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Matsumoto T., Beach D. Premature initiation of mitosis in yeast lacking RCC1 or an interacting GTPase. Cell. 1991 Jul 26;66(2):347–360. doi: 10.1016/0092-8674(91)90624-8. [DOI] [PubMed] [Google Scholar]
  32. McCully E. K., Robinow C. F. Mitosis in the fission yeast Schizosaccharomyces pombe: a comparative study with light and electron microscopy. J Cell Sci. 1971 Sep;9(2):475–507. doi: 10.1242/jcs.9.2.475. [DOI] [PubMed] [Google Scholar]
  33. Mizukami T., Chang W. I., Garkavtsev I., Kaplan N., Lombardi D., Matsumoto T., Niwa O., Kounosu A., Yanagida M., Marr T. G. A 13 kb resolution cosmid map of the 14 Mb fission yeast genome by nonrandom sequence-tagged site mapping. Cell. 1993 Apr 9;73(1):121–132. doi: 10.1016/0092-8674(93)90165-m. [DOI] [PubMed] [Google Scholar]
  34. Mohamed A. H., Chirala S. S., Mody N. H., Huang W. Y., Wakil S. J. Primary structure of the multifunctional alpha subunit protein of yeast fatty acid synthase derived from FAS2 gene sequence. J Biol Chem. 1988 Sep 5;263(25):12315–12325. [PubMed] [Google Scholar]
  35. Nishitani H., Ohtsubo M., Yamashita K., Iida H., Pines J., Yasudo H., Shibata Y., Hunter T., Nishimoto T. Loss of RCC1, a nuclear DNA-binding protein, uncouples the completion of DNA replication from the activation of cdc2 protein kinase and mitosis. EMBO J. 1991 Jun;10(6):1555–1564. doi: 10.1002/j.1460-2075.1991.tb07675.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Nurse P., Thuriaux P., Nasmyth K. Genetic control of the cell division cycle in the fission yeast Schizosaccharomyces pombe. Mol Gen Genet. 1976 Jul 23;146(2):167–178. doi: 10.1007/BF00268085. [DOI] [PubMed] [Google Scholar]
  37. Ohkura H., Yanagida M. S. pombe gene sds22+ essential for a midmitotic transition encodes a leucine-rich repeat protein that positively modulates protein phosphatase-1. Cell. 1991 Jan 11;64(1):149–157. doi: 10.1016/0092-8674(91)90216-l. [DOI] [PubMed] [Google Scholar]
  38. Pelech S. L., Sanghera J. S., Paddon H. B., Quayle K. A., Brownsey R. W. Identification of a major maturation-activated acetyl-CoA carboxylase kinase in sea star oocytes as p44mpk. Biochem J. 1991 Mar 15;274(Pt 3):759–767. doi: 10.1042/bj2740759. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Roggenkamp R., Numa S., Schweizer E. Fatty acid-requiring mutant of Saccharomyces cerevisiae defective in acetyl-CoA carboxylase. Proc Natl Acad Sci U S A. 1980 Apr;77(4):1814–1817. doi: 10.1073/pnas.77.4.1814. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Rothstein R. J. One-step gene disruption in yeast. Methods Enzymol. 1983;101:202–211. doi: 10.1016/0076-6879(83)01015-0. [DOI] [PubMed] [Google Scholar]
  41. Saka Y., Sutani T., Yamashita Y., Saitoh S., Takeuchi M., Nakaseko Y., Yanagida M. Fission yeast cut3 and cut14, members of a ubiquitous protein family, are required for chromosome condensation and segregation in mitosis. EMBO J. 1994 Oct 17;13(20):4938–4952. doi: 10.1002/j.1460-2075.1994.tb06821.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Samejima I., Matsumoto T., Nakaseko Y., Beach D., Yanagida M. Identification of seven new cut genes involved in Schizosaccharomyces pombe mitosis. J Cell Sci. 1993 May;105(Pt 1):135–143. doi: 10.1242/jcs.105.1.135. [DOI] [PubMed] [Google Scholar]
  43. 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]
  44. Sazer S., Nurse P. A fission yeast RCC1-related protein is required for the mitosis to interphase transition. EMBO J. 1994 Feb 1;13(3):606–615. doi: 10.1002/j.1460-2075.1994.tb06298.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Schweizer M., Roberts L. M., Höltke H. J., Takabayashi K., Höllerer E., Hoffmann B., Müller G., Köttig H., Schweizer E. The pentafunctional FAS1 gene of yeast: its nucleotide sequence and order of the catalytic domains. Mol Gen Genet. 1986 Jun;203(3):479–486. doi: 10.1007/BF00422073. [DOI] [PubMed] [Google Scholar]
  46. Stewart L. C., Yaffe M. P. A role for unsaturated fatty acids in mitochondrial movement and inheritance. J Cell Biol. 1991 Dec;115(5):1249–1257. doi: 10.1083/jcb.115.5.1249. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Sun G. H., Hirata A., Ohya Y., Anraku Y. Mutations in yeast calmodulin cause defects in spindle pole body functions and nuclear integrity. J Cell Biol. 1992 Dec;119(6):1625–1639. doi: 10.1083/jcb.119.6.1625. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Takahashi K., Yamada H., Yanagida M. Fission yeast minichromosome loss mutants mis cause lethal aneuploidy and replication abnormality. Mol Biol Cell. 1994 Oct;5(10):1145–1158. doi: 10.1091/mbc.5.10.1145. [DOI] [PMC free article] [PubMed] [Google Scholar]
  49. Tanaka K., Kanbe T. Mitosis in the fission yeast Schizosaccharomyces pombe as revealed by freeze-substitution electron microscopy. J Cell Sci. 1986 Feb;80:253–268. doi: 10.1242/jcs.80.1.253. [DOI] [PubMed] [Google Scholar]
  50. Uzawa S., Yanagida M. Visualization of centromeric and nucleolar DNA in fission yeast by fluorescence in situ hybridization. J Cell Sci. 1992 Feb;101(Pt 2):267–275. doi: 10.1242/jcs.101.2.267. [DOI] [PubMed] [Google Scholar]
  51. Wieland F., Renner L., Verfürth C., Lynen F. Studies on the multi-enzyme complex of yeast fatty-acid synthetase. Reversible dissociation and isolation of two polypeptide chains. Eur J Biochem. 1979 Feb 15;94(1):189–197. doi: 10.1111/j.1432-1033.1979.tb12885.x. [DOI] [PubMed] [Google Scholar]
  52. Wieland F., Siess E. A., Renner L., Verfürth C., Lynen F. Distribution of yeast fatty acid synthetase subunits: three-dimensional model of the enzyme. Proc Natl Acad Sci U S A. 1978 Dec;75(12):5792–5796. doi: 10.1073/pnas.75.12.5792. [DOI] [PMC free article] [PubMed] [Google Scholar]
  53. Wiesner P., Beck J., Beck K. F., Ripka S., Müller G., Lücke S., Schweizer E. Isolation and sequence analysis of the fatty acid synthetase FAS2 gene from Penicillium patulum. Eur J Biochem. 1988 Oct 15;177(1):69–79. doi: 10.1111/j.1432-1033.1988.tb14346.x. [DOI] [PubMed] [Google Scholar]
  54. Woods A., Munday M. R., Scott J., Yang X., Carlson M., Carling D. Yeast SNF1 is functionally related to mammalian AMP-activated protein kinase and regulates acetyl-CoA carboxylase in vivo. J Biol Chem. 1994 Jul 29;269(30):19509–19515. [PubMed] [Google Scholar]
  55. Yanagida M. Frontier questions about sister chromatid separation in anaphase. Bioessays. 1995 Jun;17(6):519–526. doi: 10.1002/bies.950170608. [DOI] [PubMed] [Google Scholar]

Articles from The Journal of Cell Biology are provided here courtesy of The Rockefeller University Press

RESOURCES