Enhancement of protein secretion in Saccharomyces cerevisiae by overproduction of Sso protein, a late-acting component of the secretory machinery

Laura Ruohonen, Jaana Toikkanen, Ville Tieaho, Mika Outola, Hans Söderlund, Sirkka Keränen (Corresponding Author)

    Research output: Contribution to journalArticleScientificpeer-review

    60 Citations (Scopus)

    Abstract

    Increased production of secreted proteins in Saccharomyces cerevisiae was achieved by overexpressing the yeast syntaxins, Sso1 or Sso2 protein, the t‐SNAREs functioning at the targeting/fusion of the Golgi‐derived secretory vesicles to the plasma membrane. Up to four‐ or six‐fold yields of a heterologous secreted protein, Bacillus α‐amylase, or an endogenous secreted protein, invertase, were obtained respectively when expressing either one of the SSO genes, SSO1 or SSO2, from the ADH1 promoter on a multicopy plasmid. Direct correlation between the Sso protein level and the amount of secreted α‐amylase was demonstrated by modulating the expression level of the SSO2 gene. Quantitation of the α‐amylase activity in the culture medium, periplasmic space and cytoplasm suggests that secretion into the periplasmic space is the primary stage at which the SSO genes exert the secretion‐enhancing function. Pulse‐chase data also support enhanced secretion efficiency obtained by SSO overexpression. Our data suggest that the Sso proteins may be rate‐limiting components of the protein secretion machinery at the exocytosis step in yeast.

    Original languageEnglish
    Pages (from-to)337 - 351
    Number of pages15
    JournalYeast
    Volume13
    Issue number4
    DOIs
    Publication statusPublished - 1997
    MoE publication typeA1 Journal article-refereed

    Fingerprint

    Secretory Component
    Yeast
    Machinery
    Saccharomyces cerevisiae
    Proteins
    Amylases
    Periplasm
    Genes
    Yeasts
    Qa-SNARE Proteins
    Saccharomyces cerevisiae Proteins
    SNARE Proteins
    beta-Fructofuranosidase
    Exocytosis
    Secretory Vesicles
    Bacillus
    Bacilli
    Cell membranes
    Culture Media
    Cytoplasm

    Cite this

    Ruohonen, Laura ; Toikkanen, Jaana ; Tieaho, Ville ; Outola, Mika ; Söderlund, Hans ; Keränen, Sirkka. / Enhancement of protein secretion in Saccharomyces cerevisiae by overproduction of Sso protein, a late-acting component of the secretory machinery. In: Yeast. 1997 ; Vol. 13, No. 4. pp. 337 - 351.
    @article{539b89b6dddc4632ab27c8c3b47b0ca6,
    title = "Enhancement of protein secretion in Saccharomyces cerevisiae by overproduction of Sso protein, a late-acting component of the secretory machinery",
    abstract = "Increased production of secreted proteins in Saccharomyces cerevisiae was achieved by overexpressing the yeast syntaxins, Sso1 or Sso2 protein, the t‐SNAREs functioning at the targeting/fusion of the Golgi‐derived secretory vesicles to the plasma membrane. Up to four‐ or six‐fold yields of a heterologous secreted protein, Bacillus α‐amylase, or an endogenous secreted protein, invertase, were obtained respectively when expressing either one of the SSO genes, SSO1 or SSO2, from the ADH1 promoter on a multicopy plasmid. Direct correlation between the Sso protein level and the amount of secreted α‐amylase was demonstrated by modulating the expression level of the SSO2 gene. Quantitation of the α‐amylase activity in the culture medium, periplasmic space and cytoplasm suggests that secretion into the periplasmic space is the primary stage at which the SSO genes exert the secretion‐enhancing function. Pulse‐chase data also support enhanced secretion efficiency obtained by SSO overexpression. Our data suggest that the Sso proteins may be rate‐limiting components of the protein secretion machinery at the exocytosis step in yeast.",
    author = "Laura Ruohonen and Jaana Toikkanen and Ville Tieaho and Mika Outola and Hans S{\"o}derlund and Sirkka Ker{\"a}nen",
    year = "1997",
    doi = "10.1002/(SICI)1097-0061(19970330)13:4<337::AID-YEA98>3.0.CO;2-K",
    language = "English",
    volume = "13",
    pages = "337 -- 351",
    journal = "Yeast",
    issn = "0749-503X",
    publisher = "Wiley",
    number = "4",

    }

    Enhancement of protein secretion in Saccharomyces cerevisiae by overproduction of Sso protein, a late-acting component of the secretory machinery. / Ruohonen, Laura; Toikkanen, Jaana; Tieaho, Ville; Outola, Mika; Söderlund, Hans; Keränen, Sirkka (Corresponding Author).

    In: Yeast, Vol. 13, No. 4, 1997, p. 337 - 351.

    Research output: Contribution to journalArticleScientificpeer-review

    TY - JOUR

    T1 - Enhancement of protein secretion in Saccharomyces cerevisiae by overproduction of Sso protein, a late-acting component of the secretory machinery

    AU - Ruohonen, Laura

    AU - Toikkanen, Jaana

    AU - Tieaho, Ville

    AU - Outola, Mika

    AU - Söderlund, Hans

    AU - Keränen, Sirkka

    PY - 1997

    Y1 - 1997

    N2 - Increased production of secreted proteins in Saccharomyces cerevisiae was achieved by overexpressing the yeast syntaxins, Sso1 or Sso2 protein, the t‐SNAREs functioning at the targeting/fusion of the Golgi‐derived secretory vesicles to the plasma membrane. Up to four‐ or six‐fold yields of a heterologous secreted protein, Bacillus α‐amylase, or an endogenous secreted protein, invertase, were obtained respectively when expressing either one of the SSO genes, SSO1 or SSO2, from the ADH1 promoter on a multicopy plasmid. Direct correlation between the Sso protein level and the amount of secreted α‐amylase was demonstrated by modulating the expression level of the SSO2 gene. Quantitation of the α‐amylase activity in the culture medium, periplasmic space and cytoplasm suggests that secretion into the periplasmic space is the primary stage at which the SSO genes exert the secretion‐enhancing function. Pulse‐chase data also support enhanced secretion efficiency obtained by SSO overexpression. Our data suggest that the Sso proteins may be rate‐limiting components of the protein secretion machinery at the exocytosis step in yeast.

    AB - Increased production of secreted proteins in Saccharomyces cerevisiae was achieved by overexpressing the yeast syntaxins, Sso1 or Sso2 protein, the t‐SNAREs functioning at the targeting/fusion of the Golgi‐derived secretory vesicles to the plasma membrane. Up to four‐ or six‐fold yields of a heterologous secreted protein, Bacillus α‐amylase, or an endogenous secreted protein, invertase, were obtained respectively when expressing either one of the SSO genes, SSO1 or SSO2, from the ADH1 promoter on a multicopy plasmid. Direct correlation between the Sso protein level and the amount of secreted α‐amylase was demonstrated by modulating the expression level of the SSO2 gene. Quantitation of the α‐amylase activity in the culture medium, periplasmic space and cytoplasm suggests that secretion into the periplasmic space is the primary stage at which the SSO genes exert the secretion‐enhancing function. Pulse‐chase data also support enhanced secretion efficiency obtained by SSO overexpression. Our data suggest that the Sso proteins may be rate‐limiting components of the protein secretion machinery at the exocytosis step in yeast.

    U2 - 10.1002/(SICI)1097-0061(19970330)13:4<337::AID-YEA98>3.0.CO;2-K

    DO - 10.1002/(SICI)1097-0061(19970330)13:4<337::AID-YEA98>3.0.CO;2-K

    M3 - Article

    VL - 13

    SP - 337

    EP - 351

    JO - Yeast

    JF - Yeast

    SN - 0749-503X

    IS - 4

    ER -