Kluyveromyces lactis SSOI and SEBI genes are functional in Saccharomyces cerevisiae and enhance production of secreted proteins when overexpressed

Jaana H. Toikkanen, Lena Sundqvist, Sirkka Keränen

Research output: Contribution to journalArticleScientificpeer-review

22 Citations (Scopus)

Abstract

The SEB1/SBH1 and the SSO genes encode components of the protein secretory machinery functioning at the opposite ends, ER translocation and exocytosis, respectively, of the secretory pathway in Saccharomyces cerevisiae. Overexpression of these genes can rescue temperature‐sensitive (ts) growth defect of many sec mutants impaired in protein secretion. Furthermore, their overexpression in wild‐type yeast enhances production of secreted proteins in S. cerevisiae, which suggests that they may be rate‐limiting factors in this process. Here we report isolation of Kluyveromyces lactis homologues of these genes. KlSSO1 and KlSEB1 were isolated as clones capable of rescuing growth of ts sso2‐1 and seb1Δ seb2Δ sem1Δ strains, respectively, at the restrictive temperature. The encoded Kluyveromyces proteins are up to 70% identical with the S. cerevisiae homologues at the amino acid level and can functionally replace them. Interestingly, KlSSO1 and KlSEB1 show similar enhancing effect on production of a secreted protein as the SSO and SEB1 genes of S. cerevisiae when overexpressed. In accordance with the high homology level of the secretory pathway proteins in different yeast species, the polyclonal antibodies raised against S. cerevisiae Seb1p, Sso2p and Sec4p can detect homologous proteins in cell lysates of K. lactis and Pichia pastoris, the latter also in Candida utilis. The GenBank Accession Nos are AF307983 (K. lactis SSO1) and AF318314 (K. lactis SEB1).
Original languageEnglish
Pages (from-to)1045-1055
JournalYeast
Volume21
Issue number12
DOIs
Publication statusPublished - 2004
MoE publication typeA1 Journal article-refereed

Fingerprint

Kluyveromyces
Yeast
Saccharomyces cerevisiae
Genes
Proteins
Secretory Pathway
Yeasts
Gene Components
Saccharomyces cerevisiae Proteins
Pichia
Nucleic Acid Databases
Exocytosis
Growth
Candida
Antibodies
Clone Cells
Machinery
Amino acids
Amino Acids
Temperature

Keywords

  • SEBI/SBHI
  • SSO genes
  • Kluyveromyces lactis
  • complementation
  • protein secretion
  • increased protein production
  • multicopy suppression
  • Saccharomyces cerevisiae

Cite this

Toikkanen, Jaana H. ; Sundqvist, Lena ; Keränen, Sirkka. / Kluyveromyces lactis SSOI and SEBI genes are functional in Saccharomyces cerevisiae and enhance production of secreted proteins when overexpressed. In: Yeast. 2004 ; Vol. 21, No. 12. pp. 1045-1055.
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abstract = "The SEB1/SBH1 and the SSO genes encode components of the protein secretory machinery functioning at the opposite ends, ER translocation and exocytosis, respectively, of the secretory pathway in Saccharomyces cerevisiae. Overexpression of these genes can rescue temperature‐sensitive (ts) growth defect of many sec mutants impaired in protein secretion. Furthermore, their overexpression in wild‐type yeast enhances production of secreted proteins in S. cerevisiae, which suggests that they may be rate‐limiting factors in this process. Here we report isolation of Kluyveromyces lactis homologues of these genes. KlSSO1 and KlSEB1 were isolated as clones capable of rescuing growth of ts sso2‐1 and seb1Δ seb2Δ sem1Δ strains, respectively, at the restrictive temperature. The encoded Kluyveromyces proteins are up to 70{\%} identical with the S. cerevisiae homologues at the amino acid level and can functionally replace them. Interestingly, KlSSO1 and KlSEB1 show similar enhancing effect on production of a secreted protein as the SSO and SEB1 genes of S. cerevisiae when overexpressed. In accordance with the high homology level of the secretory pathway proteins in different yeast species, the polyclonal antibodies raised against S. cerevisiae Seb1p, Sso2p and Sec4p can detect homologous proteins in cell lysates of K. lactis and Pichia pastoris, the latter also in Candida utilis. The GenBank Accession Nos are AF307983 (K. lactis SSO1) and AF318314 (K. lactis SEB1).",
keywords = "SEBI/SBHI, SSO genes, Kluyveromyces lactis, complementation, protein secretion, increased protein production, multicopy suppression, Saccharomyces cerevisiae",
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Kluyveromyces lactis SSOI and SEBI genes are functional in Saccharomyces cerevisiae and enhance production of secreted proteins when overexpressed. / Toikkanen, Jaana H.; Sundqvist, Lena; Keränen, Sirkka.

In: Yeast, Vol. 21, No. 12, 2004, p. 1045-1055.

Research output: Contribution to journalArticleScientificpeer-review

TY - JOUR

T1 - Kluyveromyces lactis SSOI and SEBI genes are functional in Saccharomyces cerevisiae and enhance production of secreted proteins when overexpressed

AU - Toikkanen, Jaana H.

AU - Sundqvist, Lena

AU - Keränen, Sirkka

PY - 2004

Y1 - 2004

N2 - The SEB1/SBH1 and the SSO genes encode components of the protein secretory machinery functioning at the opposite ends, ER translocation and exocytosis, respectively, of the secretory pathway in Saccharomyces cerevisiae. Overexpression of these genes can rescue temperature‐sensitive (ts) growth defect of many sec mutants impaired in protein secretion. Furthermore, their overexpression in wild‐type yeast enhances production of secreted proteins in S. cerevisiae, which suggests that they may be rate‐limiting factors in this process. Here we report isolation of Kluyveromyces lactis homologues of these genes. KlSSO1 and KlSEB1 were isolated as clones capable of rescuing growth of ts sso2‐1 and seb1Δ seb2Δ sem1Δ strains, respectively, at the restrictive temperature. The encoded Kluyveromyces proteins are up to 70% identical with the S. cerevisiae homologues at the amino acid level and can functionally replace them. Interestingly, KlSSO1 and KlSEB1 show similar enhancing effect on production of a secreted protein as the SSO and SEB1 genes of S. cerevisiae when overexpressed. In accordance with the high homology level of the secretory pathway proteins in different yeast species, the polyclonal antibodies raised against S. cerevisiae Seb1p, Sso2p and Sec4p can detect homologous proteins in cell lysates of K. lactis and Pichia pastoris, the latter also in Candida utilis. The GenBank Accession Nos are AF307983 (K. lactis SSO1) and AF318314 (K. lactis SEB1).

AB - The SEB1/SBH1 and the SSO genes encode components of the protein secretory machinery functioning at the opposite ends, ER translocation and exocytosis, respectively, of the secretory pathway in Saccharomyces cerevisiae. Overexpression of these genes can rescue temperature‐sensitive (ts) growth defect of many sec mutants impaired in protein secretion. Furthermore, their overexpression in wild‐type yeast enhances production of secreted proteins in S. cerevisiae, which suggests that they may be rate‐limiting factors in this process. Here we report isolation of Kluyveromyces lactis homologues of these genes. KlSSO1 and KlSEB1 were isolated as clones capable of rescuing growth of ts sso2‐1 and seb1Δ seb2Δ sem1Δ strains, respectively, at the restrictive temperature. The encoded Kluyveromyces proteins are up to 70% identical with the S. cerevisiae homologues at the amino acid level and can functionally replace them. Interestingly, KlSSO1 and KlSEB1 show similar enhancing effect on production of a secreted protein as the SSO and SEB1 genes of S. cerevisiae when overexpressed. In accordance with the high homology level of the secretory pathway proteins in different yeast species, the polyclonal antibodies raised against S. cerevisiae Seb1p, Sso2p and Sec4p can detect homologous proteins in cell lysates of K. lactis and Pichia pastoris, the latter also in Candida utilis. The GenBank Accession Nos are AF307983 (K. lactis SSO1) and AF318314 (K. lactis SEB1).

KW - SEBI/SBHI

KW - SSO genes

KW - Kluyveromyces lactis

KW - complementation

KW - protein secretion

KW - increased protein production

KW - multicopy suppression

KW - Saccharomyces cerevisiae

U2 - 10.1002/yea.1151

DO - 10.1002/yea.1151

M3 - Article

VL - 21

SP - 1045

EP - 1055

JO - Yeast

JF - Yeast

SN - 0749-503X

IS - 12

ER -