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J Biol Chem, Vol. 274, Issue 16, 11296-11302, April 16, 1999

Disruption of YHC8, a Member of the TSR1 Gene Family, Reveals Its Direct Involvement in Yeast Protein Translocation

Choukri Ben Mamoun^§, Jean-Marie Beckerich, Claude Gaillardin, and Francois Kepes

From the  Laboratoire de Génétique Moléculaire et Cellulaire, INRA-CNRS, Centre de Biotechnologie Agro Industrielle, Institut National Agronomique Paris-Grignon, 78850 Thiverval-Grignon, France, ^§ Departments of Molecular Microbiology and Medicine, Washington University School of Medicine, St. Louis, Missouri 63110,  Service de Biochimie et de Génétique Moléculaire, DBCM/DSV, BAT.142, CEA/Saclay, 91191 Gif Cedex, France

Genetic studies of Saccharomyces cerevisiae have identified many components acting to deliver specific proteins to their cellular locations. Genome analysis, however, has indicated that additional genes may also participate in such protein trafficking. The product of the yeast Yarrowia lipolytica TSR1 gene promotes the signal recognition particle-dependent translocation of secretory proteins through the endoplasmic reticulum. Here we describe the identification of a new gene family of proteins that is well conserved among different yeast species. The TSR1 genes encode polypeptides that share the same protein domain distribution and, like Tsr1p, may play an important role in the early steps of the signal recognition particle-dependent translocation pathway. We have identified five homologues of the TSR1 gene, four of them from the yeast Saccharomyces cerevisiae and the other from Hansenula polymorpha. We generated a null mutation in the S. cerevisiae YHC8 gene, the closest homologue to Y. lipolytica TSR1, and used different soluble (carboxypeptidase Y, -factor, invertase) and membrane (dipeptidyl-aminopeptidase) secretory proteins to study its phenotype. A large accumulation of soluble protein precursors was detected in the mutant strain. Immunofluorescence experiments show that Yhc8p is localized in the endoplasmic reticulum. We propose that the YHC8 gene is a new and important component of the S. cerevisiae endoplasmic reticulum membrane and that it functions in protein translocation/insertion of secretory proteins through or into this compartment.

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