Bph1p, the Saccharomyces cerevisiae homologue of CHS1/beige, functions in cell wall formation and protein sorting.
Mutations in the Chediak-Higashi syndrome gene (CHS1) and its murine homologue Beige result in the formation of enlarged lysosomes. BPH1 (Beige Protein Homologue 1) encodes the Saccharomyces cerevisiae homologue of CHS1/Beige. BPH1 is not essential and the encoded protein was found to be both cytosolic and peripherally bound to a membrane. Neither disruption nor overexpression of BPH1 affected vacuole morphology as assessed by fluorescence microscopy. The deltabph1 strain showed an impaired growth on defined synthetic media containing potassium acetate buffered below pH 4.25, increased sensitivity to calcofluor white, and increased agglutination in response to low pH. A library screen identified VPS9, FLO1, FLO9, BTS1 and OKP1 as high copy suppressors of the growth defect of deltabph1 on both low pH potassium acetate and calcofluor white. The deltabph1 strain demonstrated a mild defect in sorting vacuolar components, including increased secretion of carboxypeptidase Y and missorting of alkaline phosphatase. Overexpression of VPS9, BTS1 and OKP1 suppressed the carboxypeptidase Y secretion defect of deltabph1. Overexpression of BPH1 was found to suppress the calcofluor white sensitivity of a class E VPS deletion strain, deltavta1. Together, these data suggest that Bph1p associates with a membrane and is involved in protein sorting and cell wall formation.
Department of Pathology, Division of Cell Biology and Immunology, University of Utah Health Science Center, Salt Lake City, Utah 84132, USA.
Shelly L
SL
Shiflett
Michael B
MB
Vaughn
Dinh
D
Huynh
Jerry
J
Kaplan
Diane McVey
DM
Ward
D000595:Amino Acid Sequence; D001557:Benzenesulfonates; D002473:Cell Wall / Q000187:drug effects / Q000235:genetics / Q000502:physiology*; D005456:Fluorescent Dyes; D008969:Molecular Sequence Data; D009154:Mutation; D021381:Protein Transport / Q000235:genetics / Q000502:physiology; D011506:Proteins / Q000235:genetics* / Q000502:physiology; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000502:physiology*; D033921:Vesicular Transport Proteins
D001557:Benzenesulfonates; C497815:Bph1 protein, S cerevisiae; D005456:Fluorescent Dyes; C102161:LYST protein, human; D011506:Proteins; D029701:Saccharomyces cerevisiae Proteins; D033921:Vesicular Transport Proteins; C007061:C.I. Fluorescent Brightening Agent 28