title: Reduction of BiP levels decreases heterologous protein secretion in Saccharomyces cerevisiae.
abstract: Increased levels of the endoplasmic reticulum-resident protein folding chaperone BiP would be expected to either increase protein secretory capacity by improved solubilization of folding precursors or decrease secretory capacity by binding and retaining misfolded proteins. To address this question, the relationship between BiP levels and heterologous secretion in yeast was determined. A yeast strain was constructed in which BiP expression is tunable from 5 to 250% of wild-type levels, and this strain was used to explore the effect of varying BiP level on overall secretion of three heterologous proteins: human granulocyte colony-stimulating factor, Schizosaccharomyces pombe acid phosphatase, and bovine pancreatic trypsin inhibitor. For all three proteins examined, reduction in BiP expression below wild-type level diminished overall secretion, whereas 5-fold BiP overexpression from a constitutive glycolytic promoter did not substantially increase or decrease secretion titers. These results are consistent with a positive role for BiP in promoting membrane translocation and solubilization of folding precursors but are inconsistent with a negative role in proofreading and improper retention of heterologous secreted proteins.
authors.affiliation: Department of Chemical Engineering, University of Illinois at Urbana-Champaign, Urbana, Illinois 61801, USA.
authors.forename: A S
authors.initials: AS
authors.lastname: Robinson
authors.forename: J A
authors.initials: JA
authors.lastname: Bockhaus
authors.forename: A C
authors.initials: AC
authors.lastname: Voegler
authors.forename: K D
authors.initials: KD
authors.lastname: Wittrup
mesh_terms: D000818:Animals; D007611:Aprotinin / Q000378:metabolism; D002417:Cattle; D017931:DNA Primers / Q000737:chemistry; D005656:Fungal Proteins / Q000378:metabolism*; D018840:HSP70 Heat-Shock Proteins / Q000378:metabolism*; D017510:Protein Folding; D011994:Recombinant Proteins / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000378:metabolism*; D012568:Schizosaccharomyces
chemical_list: D017931:DNA Primers; D005656:Fungal Proteins; D018840:HSP70 Heat-Shock Proteins; C060153:KAR2 protein, yeast; D011994:Recombinant Proteins; D007611:Aprotinin