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. 1993 Jun;175(12):3823–3837. doi: 10.1128/jb.175.12.3823-3837.1993

SSG1, a gene encoding a sporulation-specific 1,3-beta-glucanase in Saccharomyces cerevisiae.

P San Segundo 1, J Correa 1, C R Vazquez de Aldana 1, F del Rey 1
PMCID: PMC204799  PMID: 8509335

Abstract

In Saccharomyces cerevisiae, the meiotic process is accompanied by a large increase in 1,3-beta-glucan-degradative activity. The molecular cloning of the gene (SSG1) encoding a sporulation-specific exo-1,3-beta-glucanase was achieved by screening a genomic library with a DNA probe obtained by polymerase chain reaction amplification using synthetic oligonucleotides designed according to the nucleotide sequence predicted from the amino-terminal region of the purified protein. DNA sequencing indicates that the SSG1 gene specifies a 445-amino-acid polypeptide (calculated molecular mass, 51.8 kDa) showing extensive similarity to the extracellular exo-1,3-beta-glucanases encoded by the EXG1 gene (C. R. Vazquez de Aldana, J. Correa, P. San Segundo, A. Bueno, A. R. Nebreda, E. Mendez, and F. del Rey, Gene 97:173-182, 1991). The N-terminal domain of the putative precursor is a very hydrophobic segment with structural features resembling those of signal peptides of secreted proteins. Northern (RNA) analysis reveals a unique SSG1-specific transcript, 1.7 kb long, which can be detected only in sporulating diploids (MATa/MAT alpha) but does not appear in vegetatively growing cells or in nonsporulating diploids (MAT alpha/MAT alpha) when incubated under nitrogen starvation conditions. The meiotic time course of SSG1 induction indicates that the gene is transcribed only in the late stages of the process, beginning at the time of meiosis I and reaching a maximum during spore formation. Homozygous ssg1/ssg1 mutant diploids are able to complete sporulation, although with a significant delay in the appearance of mature asci.

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Selected References

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

  1. Baker B. S., Carpenter A. T., Esposito M. S., Esposito R. E., Sandler L. The genetic control of meiosis. Annu Rev Genet. 1976;10:53–134. doi: 10.1146/annurev.ge.10.120176.000413. [DOI] [PubMed] [Google Scholar]
  2. Beckett A., Illingworth R. F., Rose A. H. Ascospore wall development in Saccharomyces cerevisiae. J Bacteriol. 1973 Feb;113(2):1054–1057. doi: 10.1128/jb.113.2.1054-1057.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bennetzen J. L., Hall B. D. Codon selection in yeast. J Biol Chem. 1982 Mar 25;257(6):3026–3031. [PubMed] [Google Scholar]
  4. Bennetzen J. L., Hall B. D. The primary structure of the Saccharomyces cerevisiae gene for alcohol dehydrogenase. J Biol Chem. 1982 Mar 25;257(6):3018–3025. [PubMed] [Google Scholar]
  5. Boeke J. D., Trueheart J., Natsoulis G., Fink G. R. 5-Fluoroorotic acid as a selective agent in yeast molecular genetics. Methods Enzymol. 1987;154:164–175. doi: 10.1016/0076-6879(87)54076-9. [DOI] [PubMed] [Google Scholar]
  6. Buckingham L. E., Wang H. T., Elder R. T., McCarroll R. M., Slater M. R., Esposito R. E. Nucleotide sequence and promoter analysis of SPO13, a meiosis-specific gene of Saccharomyces cerevisiae. Proc Natl Acad Sci U S A. 1990 Dec;87(23):9406–9410. doi: 10.1073/pnas.87.23.9406. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Cigan A. M., Donahue T. F. Sequence and structural features associated with translational initiator regions in yeast--a review. Gene. 1987;59(1):1–18. doi: 10.1016/0378-1119(87)90261-7. [DOI] [PubMed] [Google Scholar]
  8. Clancy M. J., Buten-Magee B., Straight D. J., Kennedy A. L., Partridge R. M., Magee P. T. Isolation of genes expressed preferentially during sporulation in the yeast Saccharomyces cerevisiae. Proc Natl Acad Sci U S A. 1983 May;80(10):3000–3004. doi: 10.1073/pnas.80.10.3000. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Clancy M. J., Smith L. M., Magee P. T. Developmental regulation of a sporulation-specific enzyme activity in Saccharomyces cerevisiae. Mol Cell Biol. 1982 Feb;2(2):171–178. doi: 10.1128/mcb.2.2.171. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Colonna W. J., Magee P. T. Glycogenolytic enzymes in sporulating yeast. J Bacteriol. 1978 Jun;134(3):844–853. doi: 10.1128/jb.134.3.844-853.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. De Loose M., Alliotte T., Gheysen G., Genetello C., Gielen J., Soetaert P., Van Montagu M., Inzé D. Primary structure of a hormonally regulated beta-glucanase of Nicotiana plumbaginifolia. Gene. 1988 Oct 15;70(1):13–23. doi: 10.1016/0378-1119(88)90100-x. [DOI] [PubMed] [Google Scholar]
  12. Dobson M. J., Tuite M. F., Roberts N. A., Kingsman A. J., Kingsman S. M., Perkins R. E., Conroy S. C., Fothergill L. A. Conservation of high efficiency promoter sequences in Saccharomyces cerevisiae. Nucleic Acids Res. 1982 Apr 24;10(8):2625–2637. doi: 10.1093/nar/10.8.2625. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Edman P., Begg G. A protein sequenator. Eur J Biochem. 1967 Mar;1(1):80–91. doi: 10.1007/978-3-662-25813-2_14. [DOI] [PubMed] [Google Scholar]
  14. Esposito M. S., Esposito R. E. Genes controlling meiosis and spore formation in yeast. Genetics. 1974 Sep;78(1):215–225. doi: 10.1093/genetics/78.1.215. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Farkas V., Biely P., Bauer S. Extracellular beta-glucanases of the yeast, Saccharomyces cerevisiae. Biochim Biophys Acta. 1973 Sep 15;321(1):246–255. doi: 10.1016/0005-2744(73)90079-x. [DOI] [PubMed] [Google Scholar]
  16. Feinberg A. P., Vogelstein B. A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem. 1983 Jul 1;132(1):6–13. doi: 10.1016/0003-2697(83)90418-9. [DOI] [PubMed] [Google Scholar]
  17. Fincher G. B., Lock P. A., Morgan M. M., Lingelbach K., Wettenhall R. E., Mercer J. F., Brandt A., Thomsen K. K. Primary structure of the (1-->3,1-->4)-beta-D-glucan 4-glucohydrolase from barley aleurone. Proc Natl Acad Sci U S A. 1986 Apr;83(7):2081–2085. doi: 10.1073/pnas.83.7.2081. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Garber A. T., Segall J. The SPS4 gene of Saccharomyces cerevisiae encodes a major sporulation-specific mRNA. Mol Cell Biol. 1986 Dec;6(12):4478–4485. doi: 10.1128/mcb.6.12.4478. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Gottlin-Ninfa E., Kaback D. B. Isolation and functional analysis of sporulation-induced transcribed sequences from Saccharomyces cerevisiae. Mol Cell Biol. 1986 Jun;6(6):2185–2197. doi: 10.1128/mcb.6.6.2185. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Henikoff S. Unidirectional digestion with exonuclease III creates targeted breakpoints for DNA sequencing. Gene. 1984 Jun;28(3):351–359. doi: 10.1016/0378-1119(84)90153-7. [DOI] [PubMed] [Google Scholar]
  21. Hien N. H., Fleet G. H. Separation and characterization of six (1 leads to 3)-beta-glucanases from Saccharomyces cerevisiae. J Bacteriol. 1983 Dec;156(3):1204–1213. doi: 10.1128/jb.156.3.1204-1213.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Holaway B. L., Lehman D. J., Primerano D. A., Magee P. T., Clancy M. J. Sporulation-regulated genes of Saccharomyces cerevisiae. Curr Genet. 1985;10(3):163–169. doi: 10.1007/BF00798745. [DOI] [PubMed] [Google Scholar]
  23. Hopper A. K., Magee P. T., Welch S. K., Friedman M., Hall B. D. Macromolecule synthesis and breakdown in relation to sporulation and meiosis in yeast. J Bacteriol. 1974 Aug;119(2):619–628. doi: 10.1128/jb.119.2.619-628.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Hubbard S. C., Ivatt R. J. Synthesis and processing of asparagine-linked oligosaccharides. Annu Rev Biochem. 1981;50:555–583. doi: 10.1146/annurev.bi.50.070181.003011. [DOI] [PubMed] [Google Scholar]
  25. 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]
  26. Kaback D. B., Feldberg L. R. Saccharomyces cerevisiae exhibits a sporulation-specific temporal pattern of transcript accumulation. Mol Cell Biol. 1985 Apr;5(4):751–761. doi: 10.1128/mcb.5.4.751. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Kallal L. A., Bhattacharyya M., Grove S. N., Iannacone R. F., Pugh T. A., Primerano D. A., Clancy M. J. Functional analysis of the sporulation-specific SPR6 gene of Saccharomyces cerevisiae. Curr Genet. 1990 Nov;18(4):293–301. doi: 10.1007/BF00318210. [DOI] [PubMed] [Google Scholar]
  28. Kane S. M., Roth R. Carbohydrate metabolism during ascospore development in yeast. J Bacteriol. 1974 Apr;118(1):8–14. doi: 10.1128/jb.118.1.8-14.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Kao G., Mannix D. G., Holaway B. L., Finn M. C., Bonny A. E., Clancy M. J. Dependence of inessential late gene expression on early meiotic events in Saccharomyces cerevisiae. Mol Gen Genet. 1989 Feb;215(3):490–500. doi: 10.1007/BF00427048. [DOI] [PubMed] [Google Scholar]
  30. Klebl F., Tanner W. Molecular cloning of a cell wall exo-beta-1,3-glucanase from Saccharomyces cerevisiae. J Bacteriol. 1989 Nov;171(11):6259–6264. doi: 10.1128/jb.171.11.6259-6264.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Kozak M. The scanning model for translation: an update. J Cell Biol. 1989 Feb;108(2):229–241. doi: 10.1083/jcb.108.2.229. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Kurtz S., Lindquist S. Changing patterns of gene expression during sporulation in yeast. Proc Natl Acad Sci U S A. 1984 Dec;81(23):7323–7327. doi: 10.1073/pnas.81.23.7323. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Kurtz S., Lindquist S. Subcellular differentiation in sporulating yeast cells. Cell. 1986 Jun 6;45(5):771–779. doi: 10.1016/0092-8674(86)90791-9. [DOI] [PubMed] [Google Scholar]
  34. Kyte J., Doolittle R. F. A simple method for displaying the hydropathic character of a protein. J Mol Biol. 1982 May 5;157(1):105–132. doi: 10.1016/0022-2836(82)90515-0. [DOI] [PubMed] [Google Scholar]
  35. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  36. Law D. T., Segall J. The SPS100 gene of Saccharomyces cerevisiae is activated late in the sporulation process and contributes to spore wall maturation. Mol Cell Biol. 1988 Feb;8(2):912–922. doi: 10.1128/mcb.8.2.912. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Lynn R. R., Magee P. T. Development of the spore wall during ascospore formation in Saccharomyces cerevisiae. J Cell Biol. 1970 Mar;44(3):688–692. doi: 10.1083/jcb.44.3.688. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Morrissey J. H. Silver stain for proteins in polyacrylamide gels: a modified procedure with enhanced uniform sensitivity. Anal Biochem. 1981 Nov 1;117(2):307–310. doi: 10.1016/0003-2697(81)90783-1. [DOI] [PubMed] [Google Scholar]
  39. Nebreda A. R., Vazquez C. R., Villa T. G., Villanueva J. R., del Rey F. Heterogeneous glycosylation of the EXG1 gene product accounts for the two extracellular exo-beta-glucanases of Saccharomyces cerevisiae. FEBS Lett. 1987 Aug 10;220(1):27–30. doi: 10.1016/0014-5793(87)80869-4. [DOI] [PubMed] [Google Scholar]
  40. Nebreda A. R., Villa T. G., Villanueva J. R., del Rey F. Cloning of genes related to exo-beta-glucanase production in Saccharomyces cerevisiae: characterization of an exo-beta-glucanase structural gene. Gene. 1986;47(2-3):245–259. doi: 10.1016/0378-1119(86)90068-5. [DOI] [PubMed] [Google Scholar]
  41. Ng R., Abelson J. Isolation and sequence of the gene for actin in Saccharomyces cerevisiae. Proc Natl Acad Sci U S A. 1980 Jul;77(7):3912–3916. doi: 10.1073/pnas.77.7.3912. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Nombela C., Molina M., Cenamor R., Sanchez M. Yeast beta-glucanases: a complex system of secreted enzymes. Microbiol Sci. 1988 Nov;5(11):328–332. [PubMed] [Google Scholar]
  43. Percival-Smith A., Segall J. Characterization and mutational analysis of a cluster of three genes expressed preferentially during sporulation of Saccharomyces cerevisiae. Mol Cell Biol. 1986 Jul;6(7):2443–2451. doi: 10.1128/mcb.6.7.2443. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Percival-Smith A., Segall J. Isolation of DNA sequences preferentially expressed during sporulation in Saccharomyces cerevisiae. Mol Cell Biol. 1984 Jan;4(1):142–150. doi: 10.1128/mcb.4.1.142. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Peterson G. L. A simplification of the protein assay method of Lowry et al. which is more generally applicable. Anal Biochem. 1977 Dec;83(2):346–356. doi: 10.1016/0003-2697(77)90043-4. [DOI] [PubMed] [Google Scholar]
  46. Ponticelli A. S., Struhl K. Analysis of Saccharomyces cerevisiae his3 transcription in vitro: biochemical support for multiple mechanisms of transcription. Mol Cell Biol. 1990 Jun;10(6):2832–2839. doi: 10.1128/mcb.10.6.2832. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Proudfoot N. J., Brownlee G. G. 3' non-coding region sequences in eukaryotic messenger RNA. Nature. 1976 Sep 16;263(5574):211–214. doi: 10.1038/263211a0. [DOI] [PubMed] [Google Scholar]
  48. Ramírez M., Hernández L. M., Larriba G. A similar protein portion for two exoglucanases secreted by Saccharomyces cerevisiae. Arch Microbiol. 1989;151(5):391–398. doi: 10.1007/BF00416596. [DOI] [PubMed] [Google Scholar]
  49. Rose M. D., Novick P., Thomas J. H., Botstein D., Fink G. R. A Saccharomyces cerevisiae genomic plasmid bank based on a centromere-containing shuttle vector. Gene. 1987;60(2-3):237–243. doi: 10.1016/0378-1119(87)90232-0. [DOI] [PubMed] [Google Scholar]
  50. 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]
  51. SMOGYI M. Notes on sugar determination. J Biol Chem. 1952 Mar;195(1):19–23. [PubMed] [Google Scholar]
  52. 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]
  53. Shinshi H., Wenzler H., Neuhaus J. M., Felix G., Hofsteenge J., Meins F. Evidence for N- and C-terminal processing of a plant defense-related enzyme: Primary structure of tobacco prepro-beta-1,3-glucanase. Proc Natl Acad Sci U S A. 1988 Aug;85(15):5541–5545. doi: 10.1073/pnas.85.15.5541. [DOI] [PMC free article] [PubMed] [Google Scholar]
  54. Stotz A., Linder P. The ADE2 gene from Saccharomyces cerevisiae: sequence and new vectors. Gene. 1990 Oct 30;95(1):91–98. doi: 10.1016/0378-1119(90)90418-q. [DOI] [PubMed] [Google Scholar]
  55. Struhl K. Molecular mechanisms of transcriptional regulation in yeast. Annu Rev Biochem. 1989;58:1051–1077. doi: 10.1146/annurev.bi.58.070189.005155. [DOI] [PubMed] [Google Scholar]
  56. Struhl K., Stinchcomb D. T., Scherer S., Davis R. W. High-frequency transformation of yeast: autonomous replication of hybrid DNA molecules. Proc Natl Acad Sci U S A. 1979 Mar;76(3):1035–1039. doi: 10.1073/pnas.76.3.1035. [DOI] [PMC free article] [PubMed] [Google Scholar]
  57. Sánchez A., Villanueva J. R., Villa T. G. Saccharomyces cerevisiae secretes 2 exo-beta-glucanases. FEBS Lett. 1982 Feb 22;138(2):209–212. doi: 10.1016/0014-5793(82)80443-2. [DOI] [PubMed] [Google Scholar]
  58. Vazquez de Aldana C. R., Correa J., San Segundo P., Bueno A., Nebreda A. R., Mendez E., del Rey F. Nucleotide sequence of the exo-1,3-beta-glucanase-encoding gene, EXG1, of the yeast Saccharomyces cerevisiae. Gene. 1991 Jan 15;97(2):173–182. doi: 10.1016/0378-1119(91)90049-h. [DOI] [PubMed] [Google Scholar]
  59. Winston F., Chumley F., Fink G. R. Eviction and transplacement of mutant genes in yeast. Methods Enzymol. 1983;101:211–228. doi: 10.1016/0076-6879(83)01016-2. [DOI] [PubMed] [Google Scholar]
  60. Yamashita I., Fukui S. Transcriptional control of the sporulation-specific glucoamylase gene in the yeast Saccharomyces cerevisiae. Mol Cell Biol. 1985 Nov;5(11):3069–3073. doi: 10.1128/mcb.5.11.3069. [DOI] [PMC free article] [PubMed] [Google Scholar]
  61. Yamashita I., Maemura T., Hatano T., Fukui S. Polymorphic extracellular glucoamylase genes and their evolutionary origin in the yeast Saccharomyces diastaticus. J Bacteriol. 1985 Feb;161(2):574–582. doi: 10.1128/jb.161.2.574-582.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  62. Yamashita I., Nakamura M., Fukui S. Gene fusion is a possible mechanism underlying the evolution of STA1. J Bacteriol. 1987 May;169(5):2142–2149. doi: 10.1128/jb.169.5.2142-2149.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  63. Zaret K. S., Sherman F. DNA sequence required for efficient transcription termination in yeast. Cell. 1982 Mar;28(3):563–573. doi: 10.1016/0092-8674(82)90211-2. [DOI] [PubMed] [Google Scholar]
  64. Zonneveld B. J. Morphogenesis in Aspergillus nidulans. The significance of a alpha-1, 3-glucan of the cell wall and alpha-1, 3-glucanase for cleistothecium development. Biochim Biophys Acta. 1972 Jun 26;273(1):174–187. doi: 10.1016/0304-4165(72)90205-x. [DOI] [PubMed] [Google Scholar]
  65. del Rey F., Santos T., García-Acha I., Nombela C. Synthesis of 1,3-beta-glucanases in Saccharomyces cerevisiae during the mitotic cycle, mating, and sporulation. J Bacteriol. 1979 Sep;139(3):924–931. doi: 10.1128/jb.139.3.924-931.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  66. del Rey F., Santos T., García-Acha I., Nombela C. Synthesis of beta-glucanases during sporulation in Saccharomyces cerevisiae: formation of a new, sporulation-specific 1,3-beta-glucanase. J Bacteriol. 1980 Aug;143(2):621–627. doi: 10.1128/jb.143.2.621-627.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  67. von Heijne G. Patterns of amino acids near signal-sequence cleavage sites. Eur J Biochem. 1983 Jun 1;133(1):17–21. doi: 10.1111/j.1432-1033.1983.tb07424.x. [DOI] [PubMed] [Google Scholar]

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