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J Cell Biol. 1990 Oct 1; 111(4): 1451–1464.
doi: 10.1083/jcb.111.4.1451
PMCID: PMC2116254
PMID: 2211820

The SPA2 gene of Saccharomyces cerevisiae is important for pheromone- induced morphogenesis and efficient mating

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Abstract

Upon exposure to mating pheromone, Saccharomyces cerevisiae undergoes cellular differentiation to form a morphologically distinct cell called a "shmoo". Double staining experiments revealed that both the SPA2 protein and actin localize to the shmoo tip which is the site of polarized cell growth. Actin concentrates as spots throughout the shmoo projection, while SPA2 localizes as a sharp patch at the shmoo tip. DNA sequence analysis of the SPA2 gene revealed an open reading frame 1,466 codons in length; the predicted protein sequence contains many internal repeats including a nine amino acid sequence that is imperfectly repeated 25 times. Portions of the SPA2 sequence exhibit a low-level similarity to proteins containing coiled-coil structures. Yeast cells containing a large deletion of the SPA2 gene are similar in growth rate to wild-type cells. However, spa2 mutant cells are impaired in their ability to form shmoos upon exposure to mating pheromone, and they do not mate efficiently with other spa2 mutant cells. Thus, we suggest that the SPA2 protein plays a critical role in cellular morphogenesis during mating, perhaps as a cytoskeletal protein.

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Selected References

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  • Adams AE, Pringle JR. Relationship of actin and tubulin distribution to bud growth in wild-type and morphogenetic-mutant Saccharomyces cerevisiae. J Cell Biol. 1984 Mar;98(3):934–945. [PMC free article] [PubMed] [Google Scholar]
  • Adams AE, Botstein D, Drubin DG. A yeast actin-binding protein is encoded by SAC6, a gene found by suppression of an actin mutation. Science. 1989 Jan 13;243(4888):231–233. [PubMed] [Google Scholar]
  • Baba M, Baba N, Ohsumi Y, Kanaya K, Osumi M. Three-dimensional analysis of morphogenesis induced by mating pheromone alpha factor in Saccharomyces cerevisiae. J Cell Sci. 1989 Oct;94(Pt 2):207–216. [PubMed] [Google Scholar]
  • Bader BL, Magin TM, Hatzfeld M, Franke WW. Amino acid sequence and gene organization of cytokeratin no. 19, an exceptional tail-less intermediate filament protein. EMBO J. 1986 Aug;5(8):1865–1875. [PMC free article] [PubMed] [Google Scholar]
  • Conde J, Fink GR. A mutant of Saccharomyces cerevisiae defective for nuclear fusion. Proc Natl Acad Sci U S A. 1976 Oct;73(10):3651–3655. [PMC free article] [PubMed] [Google Scholar]
  • Cross F, Hartwell LH, Jackson C, Konopka JB. Conjugation in Saccharomyces cerevisiae. Annu Rev Cell Biol. 1988;4:429–457. [PubMed] [Google Scholar]
  • Drubin DG. Actin and actin-binding proteins in yeast. Cell Motil Cytoskeleton. 1990;15(1):7–11. [PubMed] [Google Scholar]
  • Drubin DG, Miller KG, Botstein D. Yeast actin-binding proteins: evidence for a role in morphogenesis. J Cell Biol. 1988 Dec;107(6 Pt 2):2551–2561. [PMC free article] [PubMed] [Google Scholar]
  • Elion EA, Grisafi PL, Fink GR. FUS3 encodes a cdc2+/CDC28-related kinase required for the transition from mitosis into conjugation. Cell. 1990 Feb 23;60(4):649–664. [PubMed] [Google Scholar]
  • Feinberg AP, Vogelstein B. A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem. 1983 Jul 1;132(1):6–13. [PubMed] [Google Scholar]
  • Haarer BK, Pringle JR. Immunofluorescence localization of the Saccharomyces cerevisiae CDC12 gene product to the vicinity of the 10-nm filaments in the mother-bud neck. Mol Cell Biol. 1987 Oct;7(10):3678–3687. [PMC free article] [PubMed] [Google Scholar]
  • Hartwell LH. Mutants of Saccharomyces cerevisiae unresponsive to cell division control by polypeptide mating hormone. J Cell Biol. 1980 Jun;85(3):811–822. [PMC free article] [PubMed] [Google Scholar]
  • Hartwell LH, Culotti J, Pringle JR, Reid BJ. Genetic control of the cell division cycle in yeast. Science. 1974 Jan 11;183(4120):46–51. [PubMed] [Google Scholar]
  • Hasek J, Rupes I, Svobodová J, Streiblová E. Tubulin and actin topology during zygote formation of Saccharomyces cerevisiae. J Gen Microbiol. 1987 Dec;133(12):3355–3363. [PubMed] [Google Scholar]
  • Hieter P, Mann C, Snyder M, Davis RW. Mitotic stability of yeast chromosomes: a colony color assay that measures nondisjunction and chromosome loss. Cell. 1985 Feb;40(2):381–392. [PubMed] [Google Scholar]
  • 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. [PMC free article] [PubMed] [Google Scholar]
  • Johnson DI, Pringle JR. Molecular characterization of CDC42, a Saccharomyces cerevisiae gene involved in the development of cell polarity. J Cell Biol. 1990 Jul;111(1):143–152. [PMC free article] [PubMed] [Google Scholar]
  • Johnston M, Davis RW. Sequences that regulate the divergent GAL1-GAL10 promoter in Saccharomyces cerevisiae. Mol Cell Biol. 1984 Aug;4(8):1440–1448. [PMC free article] [PubMed] [Google Scholar]
  • Karn J, Brenner S, Barnett L. Protein structural domains in the Caenorhabditis elegans unc-54 myosin heavy chain gene are not separated by introns. Proc Natl Acad Sci U S A. 1983 Jul;80(14):4253–4257. [PMC free article] [PubMed] [Google Scholar]
  • Katti SK, LeMaster DM, Eklund H. Crystal structure of thioredoxin from Escherichia coli at 1.68 A resolution. J Mol Biol. 1990 Mar 5;212(1):167–184. [PubMed] [Google Scholar]
  • Kilmartin JV, Adams AE. Structural rearrangements of tubulin and actin during the cell cycle of the yeast Saccharomyces. J Cell Biol. 1984 Mar;98(3):922–933. [PMC free article] [PubMed] [Google Scholar]
  • Kilmartin JV, Wright B, Milstein C. Rat monoclonal antitubulin antibodies derived by using a new nonsecreting rat cell line. J Cell Biol. 1982 Jun;93(3):576–582. [PMC free article] [PubMed] [Google Scholar]
  • Knapp B, Rentrop M, Schweizer J, Winter H. Nonepidermal members of the keratin multigene family: cDNA sequences and in situ localization of the mRNAs. Nucleic Acids Res. 1986 Jan 24;14(2):751–763. [PMC free article] [PubMed] [Google Scholar]
  • Konopka JB, Jenness DD, Hartwell LH. The C-terminus of the S. cerevisiae alpha-pheromone receptor mediates an adaptive response to pheromone. Cell. 1988 Aug 26;54(5):609–620. [PubMed] [Google Scholar]
  • Krieg TM, Schafer MP, Cheng CK, Filpula D, Flaherty P, Steinert PM, Roop DR. Organization of a type I keratin gene. Evidence for evolution of intermediate filaments from a common ancestral gene. J Biol Chem. 1985 May 25;260(10):5867–5870. [PubMed] [Google Scholar]
  • Landschulz WH, Johnson PF, McKnight SL. The leucine zipper: a hypothetical structure common to a new class of DNA binding proteins. Science. 1988 Jun 24;240(4860):1759–1764. [PubMed] [Google Scholar]
  • Lees JF, Shneidman PS, Skuntz SF, Carden MJ, Lazzarini RA. The structure and organization of the human heavy neurofilament subunit (NF-H) and the gene encoding it. EMBO J. 1988 Jul;7(7):1947–1955. [PMC free article] [PubMed] [Google Scholar]
  • Marchuk D, McCrohon S, Fuchs E. Complete sequence of a gene encoding a human type I keratin: sequences homologous to enhancer elements in the regulatory region of the gene. Proc Natl Acad Sci U S A. 1985 Mar;82(6):1609–1613. [PMC free article] [PubMed] [Google Scholar]
  • Molina MI, Kropp KE, Gulick J, Robbins J. The sequence of an embryonic myosin heavy chain gene and isolation of its corresponding cDNA. J Biol Chem. 1987 May 15;262(14):6478–6488. [PubMed] [Google Scholar]
  • Noble M, Lewis SA, Cowan NJ. The microtubule binding domain of microtubule-associated protein MAP1B contains a repeated sequence motif unrelated to that of MAP2 and tau. J Cell Biol. 1989 Dec;109(6 Pt 2):3367–3376. [PMC free article] [PubMed] [Google Scholar]
  • Ohya Y, Miyamoto S, Ohsumi Y, Anraku Y. Calcium-sensitive cls4 mutant of Saccharomyces cerevisiae with a defect in bud formation. J Bacteriol. 1986 Jan;165(1):28–33. [PMC free article] [PubMed] [Google Scholar]
  • Ohya Y, Ohsumi Y, Anraku Y. Isolation and characterization of Ca2+-sensitive mutants of Saccharomyces cerevisiae. J Gen Microbiol. 1986 Apr;132(4):979–988. [PubMed] [Google Scholar]
  • Pearson WR. Rapid and sensitive sequence comparison with FASTP and FASTA. Methods Enzymol. 1990;183:63–98. [PubMed] [Google Scholar]
  • Pearson WR, Lipman DJ. Improved tools for biological sequence comparison. Proc Natl Acad Sci U S A. 1988 Apr;85(8):2444–2448. [PMC free article] [PubMed] [Google Scholar]
  • Rose MD, Fink GR. KAR1, a gene required for function of both intranuclear and extranuclear microtubules in yeast. Cell. 1987 Mar 27;48(6):1047–1060. [PubMed] [Google Scholar]
  • Rothstein RJ. One-step gene disruption in yeast. Methods Enzymol. 1983;101:202–211. [PubMed] [Google Scholar]
  • Sanger F, Nicklen S, Coulson AR. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. [PMC free article] [PubMed] [Google Scholar]
  • Shenoy S, Choi JK, Bagrodia S, Copeland TD, Maller JL, Shalloway D. Purified maturation promoting factor phosphorylates pp60c-src at the sites phosphorylated during fibroblast mitosis. Cell. 1989 Jun 2;57(5):763–774. [PubMed] [Google Scholar]
  • Snyder M. The SPA2 protein of yeast localizes to sites of cell growth. J Cell Biol. 1989 Apr;108(4):1419–1429. [PMC free article] [PubMed] [Google Scholar]
  • Snyder M, Davidson N. Two gene families clustered in a small region of the Drosophila genome. J Mol Biol. 1983 May 15;166(2):101–118. [PubMed] [Google Scholar]
  • Snyder M, Davis RW. SPA1: a gene important for chromosome segregation and other mitotic functions in S. cerevisiae. Cell. 1988 Sep 9;54(6):743–754. [PubMed] [Google Scholar]
  • Teem JL, Abovich N, Kaufer NF, Schwindinger WF, Warner JR, Levy A, Woolford J, Leer RJ, van Raamsdonk-Duin MM, Mager WH, et al. A comparison of yeast ribosomal protein gene DNA sequences. Nucleic Acids Res. 1984 Nov 26;12(22):8295–8312. [PMC free article] [PubMed] [Google Scholar]
  • Simon JR, Moore PD. Homologous recombination between single-stranded DNA and chromosomal genes in Saccharomyces cerevisiae. Mol Cell Biol. 1987 Jul;7(7):2329–2334. [PMC free article] [PubMed] [Google Scholar]
  • Watzele M, Klis F, Tanner W. Purification and characterization of the inducible a agglutinin of Saccharomyces cerevisiae. EMBO J. 1988 May;7(5):1483–1488. [PMC free article] [PubMed] [Google Scholar]
  • Yanisch-Perron C, Vieira J, Messing J. Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene. 1985;33(1):103–119. [PubMed] [Google Scholar]
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