PMCCPMCCPMCC

Search tips
Search criteria 

Advanced

 
Logo of molcellbPermissionsJournals.ASM.orgJournalMCB ArticleJournal InfoAuthorsReviewers
 
Mol Cell Biol. 1991 August; 11(8): 4196–4206.
PMCID: PMC361242

The AGA1 product is involved in cell surface attachment of the Saccharomyces cerevisiae cell adhesion glycoprotein a-agglutinin.

Abstract

Saccharomyces cerevisiae a and alpha cells express the complementary cell surface glycoproteins a-agglutinin and alpha-agglutinin, respectively, which interact with one another to promote cellular aggregation during mating. Treatment of S. cerevisiae a cells with reducing agents releases the binding subunit of a-agglutinin, which has been purified and characterized; little biochemical information on the overall structure of a-agglutinin is available. To characterise a-agglutinin structure and function, we have used a genetic approach to clone an a-agglutinin structural gene (AGAI). Mutants with a-specific agglutination defects were isolated, the majority of which fell into a single complementation group, called aga1. The aga1 mutants showed wild-type pheromone production and response, efficient mating on solid medium, and a mating defect in liquid medium; these phenotypes are characteristic of agglutinin mutants. The AGA1 gene was cloned by complementation; the gene sequence indicated that it could encode a protein of 725 amino acids with high serine and threonine content, a putative N-terminal signal sequence, and a C-terminal hydrophobic sequence similar to signals for the attachment to glycosyl phosphatidylinositol anchors. Active a-agglutinin binding subunit is secreted by aga1 mutants, indicating that AGA1 is involved in cells surface attachment of a-agglutinin. This result suggests that AGA1 encodes a protein with functional similarity to the core subunits of a-agglutinin analogs from other budding yeasts. Unexpectedly, the AGA1 transcript was expressed and induced by pheromone in both a and alpha cells, suggesting that the a-specific expression of active a-agglutinin results only from a-specific regulation of the a-agglutinin binding subunit.

Full text

Full text is available as a scanned copy of the original print version. Get a printable copy (PDF file) of the complete article (2.2M), or click on a page image below to browse page by page. Links to PubMed are also available for Selected References.

Images in this article

Click on the image to see a larger version.

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Aubert JP, Biserte G, Loucheux-Lefebvre MH. Carbohydrate-peptide linkage in glycoproteins. Arch Biochem Biophys. 1976 Aug;175(2):410–418. [PubMed]
  • Boeke JD, LaCroute F, Fink GR. A positive selection for mutants lacking orotidine-5'-phosphate decarboxylase activity in yeast: 5-fluoro-orotic acid resistance. Mol Gen Genet. 1984;197(2):345–346. [PubMed]
  • Burke D, Mendonça-Previato L, Ballou CE. Cell-cell recognition in yeast: purification of Hansenula wingei 21-cell sexual agglutination factor and comparison of the factors from three genera. Proc Natl Acad Sci U S A. 1980 Jan;77(1):318–322. [PubMed]
  • Chan RK, Otte CA. Isolation and genetic analysis of Saccharomyces cerevisiae mutants supersensitive to G1 arrest by a factor and alpha factor pheromones. Mol Cell Biol. 1982 Jan;2(1):11–20. [PMC free article] [PubMed]
  • Chou PY, Fasman GD. Prediction of the secondary structure of proteins from their amino acid sequence. Adv Enzymol Relat Areas Mol Biol. 1978;47:45–148. [PubMed]
  • Costanzo MC, Mueller PP, Strick CA, Fox TD. Primary structure of wild-type and mutant alleles of the PET494 gene of Saccharomyces cerevisiae. Mol Gen Genet. 1986 Feb;202(2):294–301. [PubMed]
  • Cross F, Hartwell LH, Jackson C, Konopka JB. Conjugation in Saccharomyces cerevisiae. Annu Rev Cell Biol. 1988;4:429–457. [PubMed]
  • Cross GA. Eukaryotic protein modification and membrane attachment via phosphatidylinositol. Cell. 1987 Jan 30;48(2):179–181. [PubMed]
  • Dale RM, McClure BA, Houchins JP. A rapid single-stranded cloning strategy for producing a sequential series of overlapping clones for use in DNA sequencing: application to sequencing the corn mitochondrial 18 S rDNA. Plasmid. 1985 Jan;13(1):31–40. [PubMed]
  • Dolan JW, Kirkman C, Fields S. The yeast STE12 protein binds to the DNA sequence mediating pheromone induction. Proc Natl Acad Sci U S A. 1989 Aug;86(15):5703–5707. [PubMed]
  • Engebrecht J, Hirsch J, Roeder GS. Meiotic gene conversion and crossing over: their relationship to each other and to chromosome synapsis and segregation. Cell. 1990 Sep 7;62(5):927–937. [PubMed]
  • Errede B, Ammerer G. STE12, a protein involved in cell-type-specific transcription and signal transduction in yeast, is part of protein-DNA complexes. Genes Dev. 1989 Sep;3(9):1349–1361. [PubMed]
  • Ferguson MA, Williams AF. Cell-surface anchoring of proteins via glycosyl-phosphatidylinositol structures. Annu Rev Biochem. 1988;57:285–320. [PubMed]
  • Fiat AM, Jollès J, Aubert JP, Loucheux-Lefebvre MH, Jollès P. Localisation and importance of the sugar part of human casein. Eur J Biochem. 1980 Oct;111(2):333–339. [PubMed]
  • Gellerfors P, Axelsson K, Helander A, Johansson S, Kenne L, Lindqvist S, Pavlu B, Skottner A, Fryklund L. Isolation and characterization of a glycosylated form of human insulin-like growth factor I produced in Saccharomyces cerevisiae. J Biol Chem. 1989 Jul 5;264(19):11444–11449. [PubMed]
  • Goutte C, Johnson AD. a1 protein alters the DNA binding specificity of alpha 2 repressor. Cell. 1988 Mar 25;52(6):875–882. [PubMed]
  • Guarente L, Yocum RR, Gifford P. A GAL10-CYC1 hybrid yeast promoter identifies the GAL4 regulatory region as an upstream site. Proc Natl Acad Sci U S A. 1982 Dec;79(23):7410–7414. [PubMed]
  • Hagen DC, Sprague GF., Jr Induction of the yeast alpha-specific STE3 gene by the peptide pheromone a-factor. J Mol Biol. 1984 Oct 5;178(4):835–852. [PubMed]
  • Hagiya M, Yoshida K, Yanagishima N. The release of sex-specific substances responsible for sexual agglutination from haploid cells of Saccharomyces cerevisiae. Exp Cell Res. 1977 Feb;104(2):263–272. [PubMed]
  • Herskowitz I, Jensen RE. Putting the HO gene to work: practical uses for mating-type switching. Methods Enzymol. 1991;194:132–146. [PubMed]
  • Hill HD, Jr, Schwyzer M, Steinman HM, Hill RL. Ovine submaxillary mucin. Primary structure and peptide substrates of UDP-N-acetylgalactosamine:mucin transferase. J Biol Chem. 1977 Jun 10;252(11):3799–3804. [PubMed]
  • Hill JE, Myers AM, Koerner TJ, Tzagoloff A. Yeast/E. coli shuttle vectors with multiple unique restriction sites. Yeast. 1986 Sep;2(3):163–167. [PubMed]
  • Jensen RE, Herskowitz I. Directionality and regulation of cassette substitution in yeast. Cold Spring Harb Symp Quant Biol. 1984;49:97–104. [PubMed]
  • 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]
  • Kronstad JW, Holly JA, MacKay VL. A yeast operator overlaps an upstream activation site. Cell. 1987 Jul 31;50(3):369–377. [PubMed]
  • Kurjan J. Alpha-factor structural gene mutations in Saccharomyces cerevisiae: effects on alpha-factor production and mating. Mol Cell Biol. 1985 Apr;5(4):787–796. [PMC free article] [PubMed]
  • Kurjan J, Herskowitz I. Structure of a yeast pheromone gene (MF alpha): a putative alpha-factor precursor contains four tandem copies of mature alpha-factor. Cell. 1982 Oct;30(3):933–943. [PubMed]
  • Kyte J, Doolittle RF. A simple method for displaying the hydropathic character of a protein. J Mol Biol. 1982 May 5;157(1):105–132. [PubMed]
  • Lipke PN, Wojciechowicz D, Kurjan J. AG alpha 1 is the structural gene for the Saccharomyces cerevisiae alpha-agglutinin, a cell surface glycoprotein involved in cell-cell interactions during mating. Mol Cell Biol. 1989 Aug;9(8):3155–3165. [PMC free article] [PubMed]
  • Low MG, Saltiel AR. Structural and functional roles of glycosyl-phosphatidylinositol in membranes. Science. 1988 Jan 15;239(4837):268–275. [PubMed]
  • Marykwas DL, Fox TD. Control of the Saccharomyces cerevisiae regulatory gene PET494: transcriptional repression by glucose and translational induction by oxygen. Mol Cell Biol. 1989 Feb;9(2):484–491. [PMC free article] [PubMed]
  • McCaffrey G, Clay FJ, Kelsay K, Sprague GF., Jr Identification and regulation of a gene required for cell fusion during mating of the yeast Saccharomyces cerevisiae. Mol Cell Biol. 1987 Aug;7(8):2680–2690. [PMC free article] [PubMed]
  • Mendonça-Previato L, Burke D, Ballou CE. Sexual agglutination factors from the yeast Pichia amethionina. J Cell Biochem. 1982;19(2):171–178. [PubMed]
  • Orlean P, Ammer H, Watzele M, Tanner W. Synthesis of an O-glycosylated cell surface protein induced in yeast by alpha factor. Proc Natl Acad Sci U S A. 1986 Sep;83(17):6263–6266. [PubMed]
  • Mazella J, Poustis C, Labbe C, Checler F, Kitabgi P, Granier C, van Rietschoten J, Vincent JP. Monoiodo-[Trp11]neurotensin, a highly radioactive ligand of neurotensin receptors. Preparation, biological activity, and binding properties to rat brain synaptic membranes. J Biol Chem. 1983 Mar 25;258(6):3476–3481. [PubMed]
  • Renart J, Sandoval IV. Western blots. Methods Enzymol. 1984;104:455–460. [PubMed]
  • Rose MD, Novick P, Thomas JH, Botstein D, Fink GR. A Saccharomyces cerevisiae genomic plasmid bank based on a centromere-containing shuttle vector. Gene. 1987;60(2-3):237–243. [PubMed]
  • 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. [PubMed]
  • Taylor NW, Orton WL. Association constant of the sex-specific agglutinin in the yeast, Hansenula wingei. Biochemistry. 1970 Jul 7;9(14):2931–2934. [PubMed]
  • Taylor NW, Orton WL. Cooperation among the active binding sites in the sex-specific agglutinin from the yeast, Hansenula wingei. Biochemistry. 1971 May 25;10(11):2043–2049. [PubMed]
  • Terrance K, Heller P, Wu YS, Lipke PN. Identification of glycoprotein components of alpha-agglutinin, a cell adhesion protein from Saccharomyces cerevisiae. J Bacteriol. 1987 Feb;169(2):475–482. [PMC free article] [PubMed]
  • Terrance K, Lipke PN. Sexual agglutination in Saccharomyces cerevisiae. J Bacteriol. 1981 Dec;148(3):889–896. [PMC free article] [PubMed]
  • Terrance K, Lipke PN. Pheromone induction of agglutination in Saccharomyces cerevisiae a cells. J Bacteriol. 1987 Oct;169(10):4811–4815. [PMC free article] [PubMed]
  • Thomas PS. Hybridization of denatured RNA and small DNA fragments transferred to nitrocellulose. Proc Natl Acad Sci U S A. 1980 Sep;77(9):5201–5205. [PubMed]
  • Trueheart J, Boeke JD, Fink GR. Two genes required for cell fusion during yeast conjugation: evidence for a pheromone-induced surface protein. Mol Cell Biol. 1987 Jul;7(7):2316–2328. [PMC free article] [PubMed]
  • von Heijne G. Patterns of amino acids near signal-sequence cleavage sites. Eur J Biochem. 1983 Jun 1;133(1):17–21. [PubMed]
  • 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. [PubMed]
  • Wojciechowicz D, Lipke PN. Alpha-agglutinin expression in Saccharomyces cerevisiae. Biochem Biophys Res Commun. 1989 May 30;161(1):46–51. [PubMed]
  • Xue CB, Caldwell GA, Becker JM, Naider F. Total synthesis of the lipopeptide a-mating factor of Saccharomyces cerevisiae. Biochem Biophys Res Commun. 1989 Jul 14;162(1):253–257. [PubMed]
  • 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]
  • Yen PH, Ballou CE. Partial characterization of the sexual agglutination factor from Hansenula wingei Y-2340 type 5 cells. Biochemistry. 1974 May 21;13(11):2428–2437. [PubMed]
  • Yoshida K, HAGIYA M, Yanagishima N. Isolation and purification of the sexual agglutination substance of mating type a cells in Saccharomyces cerevisiae. Biochem Biophys Res Commun. 1976 Aug 23;71(4):1085–1094. [PubMed]

Articles from Molecular and Cellular Biology are provided here courtesy of American Society for Microbiology (ASM)