Skip to main content
SpringerLink
Log in

Over-expression ofS. cerevisiae G1 cyclins restores the viability ofalg1 N-glycosylation mutants

  • Original Paper
  • Published:
Current Genetics Aims and scope Submit manuscript

Abstract

In budding yeast, one of three G1 cyclins is required for progression though START, when cells commit to a further round of cell division. We have identified mutations inALG1 (ERC14), a gene required for N-glycosylation, which are inviable in acln1 cln2 background but are rescued by over-expression ofCLNs.CLN1 andCLN2 are much more efficient thanCLN3 in rescuing theerc14-1 allele. Theerc14-1 allele results in a significant N-glycosylation defect, and no rescue of this defect byCLN1 over-expression was detected. These data suggest thatCLN over-expression could be allowing cells to live with lower levels of N-glycosylation, possibly by overcoming a checkpoint sensitive to N-glycosylation capacity. A plasmid suppressor ofalg1, PSA1, encodes a 361 amino-acid protein with homology to NDP-hexose pyrophosphorylases, the enzymes that catalyze the formation of activated sugar nucleotides.PSA1 is an essential gene, andPSA1 transcription is nearly co-ordinately regulated withCLN2 transcription, peaking near START. Co-ordinate regulation of glycosylation, sugar nucleotide metabolism, and cell-cycle progression through G1 may be a feature that ensures adequate cell-wall precursors are present before bud emergence.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Albright CF, Robbins PW (1990) The sequence and transcript heterogeneity of the yeast gene ALGI, an essential mannosyltransferase involved in N-glycosylation. J Biol Chem 265:7042–7049

    PubMed  Google Scholar 

  • Arnold E, Tanner W (1982) An obligatory role of protein glycosylation in the life cycle of yeast cells. FEBS Lett 148:49–53

    Article  PubMed  Google Scholar 

  • Ausubel FM, Brent R, Kingston RE et al. (1987) Current protocols in molecular biology. Wiley Interscience, New York

    Google Scholar 

  • Ballou CE (1982) Yeast cell wall and cell surface. In: Strathern JN, Jones EW, Broach JR (eds) The molecular biology of the yeast Saccharomyces cerevisiae. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York, pp 335–360

    Google Scholar 

  • Benton BK, Tinkelenberg AH, Jean D, Plump SD, Cross FR (1993) Genetic analysis of Cln/Cdc28 regulation of cell morphogenesis in budding yeast. EMBO J 12:5267–5275

    PubMed  Google Scholar 

  • Breeden L, Nasmyth K (1987) Cell-cycle control of the yeastHO gene: cis- and trans-acting regulators. Cell 48:389–397

    Article  PubMed  Google Scholar 

  • Christianson TW, Sikorski RS, Dante M, Shero JH, Hieter P (1992) Multifunctional yeast high-copy-number shuttle vectors. Gene 110:119–122

    Article  PubMed  Google Scholar 

  • Couto JR, Huffaker TC, Robbins PW (1984) Cloning and expression in Escherichia coli of a yeast mannosyltransferase from the asparagine-linked glycosylation pathway. J Biol Chem 259:378–382

    PubMed  Google Scholar 

  • Cross FR (1988)DAFT, a mutant gene affecting size control, pheromone arrest, and cell-cycle kinetics ofSaccharomyces cerevisiae. Mol Cell Biol 8:4675–4684

    PubMed  Google Scholar 

  • Cross FR (1990) Cell-cycle arrest caused byCLN gene deficiency inSaccharomyces cerevisiae resembles START-I arrest and is independent of the mating-pheromone signalling pathway. Mol Cell Biol 10:6482–6490

    PubMed  Google Scholar 

  • Cross FR, Blake CM (1993) The yeast Cln3 protein is an unstable activator of Cdc28. Mol Cell Biol 13:3266–3271

    PubMed  Google Scholar 

  • Cvrcková F, Nasmyth K (1993) Yeast G1 cyclinsCLN1 and CLN2 and a GAP-like protein have a role in bud formation. EMBO J 12:5277–5286

    PubMed  Google Scholar 

  • Hartwell LH, Weinert TA (1989) Checkpoints: controls that ensure the order of cell-cycle events. Science 246:629–634

    PubMed  Google Scholar 

  • Haslik A, Tanner W (1978) Carbohydrate moiety of carboxypeptidase Y and perturbation of its biosynthesis. Eur J Biochem 91:567–575

    Article  PubMed  Google Scholar 

  • Herscovics A, Orlean P (1993) Glycoprotein biosynthesis in yeast. FASEB J 7:540–550

    PubMed  Google Scholar 

  • Huffaker TC, Robbins PW (1982) Temperature-sensitive yeast mutants deficient in asparagine-linked glycosylation. J Biol Chem 257:3203–3210

    PubMed  Google Scholar 

  • Huffaker TC, Robbins PW (1983) Yeast mutants deficient in protein glycosylation. Proc Natl Acad Sci USA 80:7466–7470

    PubMed  Google Scholar 

  • Ishii S, Volpe JJ (1990) N-linked glycoprotein synthesis and transport during G1 are necessary for astrocytic proliferation. J Neurosci Res 26:419–427

    Article  PubMed  Google Scholar 

  • Klebl F, Huffaker T, Tanner W (1984) A temperature-sensitive N-glycosylation mutant ofS. cerevisiae that behaves like a cellcycle mutant. Exp Cell Res 150:309–313

    Article  PubMed  Google Scholar 

  • Koch C, Moll T, Neuberg M, Ahorn H, Nasmyth K (1993) A role for the transcription factors Mbp1 and Swi4 in progression from G1 to S phase. Science 261:1551–1557

    PubMed  Google Scholar 

  • Kukuruzinska MA, Bergh MLE, Jackson BL (1987) Protein glycosylation in yeast. Annu Rev Biochem 56:915–944

    Article  PubMed  Google Scholar 

  • Langan TJ, Slater MC (1991) Isoprenoids and astroglial cell cycling:diminished mevalonate availability and inhibition of dolichol-linked glycoprotein synthesis arrest cycling through distinct mechanisms. J Cell Physiol 149:284–292

    Article  PubMed  Google Scholar 

  • Larsson O, Carlberg M, Zetterberg A (1993) Selective killing induced by an inhibitor of N-linked glycosylation. J Cell Sci 106:299–307

    PubMed  Google Scholar 

  • Lehle L, Tanner W (1976) The specific site of tunicamycin inhibition in the formation of dolichol-bound N-acetylglucosamine derivatives. FEBS Lett 71:167–170

    Article  Google Scholar 

  • Leung DW, Chen E, Goeddel DV (1989) A method for random mutagenesis of a defined DNA segment using a modified polymerase chain reaction. Technique 1:11–15

    Google Scholar 

  • Muhlrad D, Hunter R, Parker R (1992) A rapid method for localized mutagenesis of yeast genes. Yeast 8:79–82

    Article  PubMed  Google Scholar 

  • Nasmyth K, Dirick L (1991) The role ofSW14 andSW16 in the activity of G1 cyclins in yeast. Cell 66:995–1013

    Article  PubMed  Google Scholar 

  • Oehlen LJWM, Cross FR (1994) G1 cyclinsCLN1 andCLN2 repress the mating-factor response pathway at Start in the yeast cell cycle. Genes Dev 8:1058–1070

    PubMed  Google Scholar 

  • Ogas J, Andrews BJ, Herskowitz I (1991) Transcriptional activation ofCLN1, CLN2, and a putative new G1 cyclin (HCS26) by SW14, a positive regulator of G1-specific transcription. Cell 66:1015–1026

    Article  PubMed  Google Scholar 

  • Orlean P, Seebacher G, Tanner W (1983)Saccharomyces cerevisiae α-factor prevents formation of glycoproteins in a cells. FEBS Lett 158:247–250

    Article  PubMed  Google Scholar 

  • Orlean P, Schwaiger H, Appeltauer U, Haselbeck A, Tanner W (1984)Saccharomyces cerevisiae mating pheromones specifically inhibit the synthesis of proteins destined to be N-glycosylated. Eur J Biochem 183–189

  • Osley MA, Gould J, Kim S, Kane MY, Hereford L (1986) Identification of sequences in a yeast histone promoter involved in periodic transcription. Cell 45:537–544

    Article  PubMed  Google Scholar 

  • Patterson SI, Pate Skene JH (1994) Novel inhibitory action of tunicamycin homologues suggests a role for dynamic protein fatty acylation in growth cone-mediated neurite extension. J Cell Biol 124:521–536

    Article  PubMed  Google Scholar 

  • Pearson WR, Lipman DJ (1988) Improved tools for biological sequence comparison. Proc Natl Acad Sci USA 85:2444–2448

    Google Scholar 

  • Popolo L, Vai M, Alberghina L (1986) Identification of a glycoprotein involved in cell-cycle progression in yeast. J Biol Chem 261:3479–3482

    PubMed  Google Scholar 

  • Pringle JR, Hartwell LH (1981) The Saccharomyces cerevisiae life cycle. In: Strathern J, Jones EW, Broach JR (eds) The molecular biology of the yeast Saccharomyces cerevisiae. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York, pp 97–142

    Google Scholar 

  • Richardson HE, Wittenberg C, Cross F, Reed SI (1989) An essential G1 function for cyclin-like proteins in yeast. Cell 59:1127–1133

    Article  PubMed  Google Scholar 

  • Riles L, Dutchik JE, Baktha A, McCauley BK, Thayer EC, Leckie MP, Braden VV, Depke JE, Olson MV (1993) Physical maps of the six smallest chromosomes of Saccharomyces cerevisiae at a resolution of 2.6 kilobase pairs. Genetics 134:81–150

    PubMed  Google Scholar 

  • Roos J, Sternglanz R, Lennarz WJ (1994) A screen for yeast mutants with defects in the dolichol-mediated pathway for N-glycosylation. Proc Nat] Acad Sci USA 91:1485–1489

    Google Scholar 

  • Rothblatt JA, Meyer DI (1986) Secretion in yeast: reconstitution of the translocation and glycosylation of α-factor and invertase in a homologous cell-free system. Cell 44:619–628

    Article  PubMed  Google Scholar 

  • Sikorski RS, Hieter P (1989) A system of shuttle vectors and yeast strains designed for efficient manipulation of DNA inSaccharomyces cerevisiae. Genetics 122:19–27

    PubMed  Google Scholar 

  • Takatsuki A, Tamura G (1971) Effect of tunicamycin on the synthesis of macromolecules in cultures of chick embryo fibroblasts infected with Newcastle disease virus. J Antibiot 24:785–794

    PubMed  Google Scholar 

  • Tkacz JS, Lampen JO (1975) Tunicamycin inhibition of polyisoprenyl N-acetylglusosaminyl pyrophosphate formation in calfliver microsomes. Biochem Biophys Res Commun 65:248–257

    Article  PubMed  Google Scholar 

  • Vai M, Popolo L, Alberghina L (1987) Effect of tunicamycin on cell-cycle progression in budding yeast. Exp Cell Res 171:448–459

    Article  PubMed  Google Scholar 

  • Vai M, Gatti E, Lacana E, Popolo L, Alberghina L (1991) Isolation and deduced amino-acid sequence of the gene encoding gp115, a yeast glycophospholipid-anchored protein containing a serinerich region. J Biol Chem 266:12242–12248

    PubMed  Google Scholar 

  • Wejde J, Carlberg M, Hjertman M, Larsson O (1993) Isoprenoid regulation of cell growth: identification of mevalonate-labelled compounds inducing DNA synthesis in human breast cancer cells depleted of serum and mevalonate. J Cell Physiol 155:539–548

    Article  PubMed  Google Scholar 

Download references

Author information

Author notes

  1. Suzanne Driscoll Plump

    Present address: Department of Developmental Biology, Stanford University School of Medicine, Stanford, CA, USA

  2. Jack Roos

    Present address: Hormone Research Institute, University of California, San Francisco, CA, USA

Authors and Affiliations

  1. The Rockefeller University, 1230 York Avenue, 10021, New York, NY, USA

    Benjamin K. Benton, Suzanne Driscoll Plump & Frederick R. Cross

  2. Department of Molecular Biology and Biochemistry, The State University of New York at Stony Brook, Stony Brook, NY, USA

    Jack Roos & William J. Lennarz

Authors
  1. Benjamin K. Benton
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Suzanne Driscoll Plump
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Jack Roos
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. William J. Lennarz
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Frederick R. Cross
    View author publications

    You can also search for this author in PubMed Google Scholar

Additional information

Communicated by R.J. Rothstein

Rights and permissions

Reprints and permissions

About this article

Cite this article

Benton, B.K., Plump, S.D., Roos, J. et al. Over-expression ofS. cerevisiae G1 cyclins restores the viability ofalg1 N-glycosylation mutants. Curr Genet 29, 106–113 (1996). https://doi.org/10.1007/BF02221573

Download citation

  • Received:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF02221573

Key words

Search

Navigation