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Volume 272, Number 29, Issue of July 18, 1997 pp. 18117-18124
©1997 by The American Society for Biochemistry and Molecular Biology, Inc.

MNN6, a Member of the KRE2/MNT1 Family, Is the Gene for Mannosylphosphate Transfer in Saccharomyces cerevisiae

(Received for publication, December 23, 1996, and in revised form, May 13, 1997)

Xiao-Hui Wang , Ken-ichi Nakayama , Yoh-ichi Shimma , Atsushi Tanaka ^§ and Yoshifumi Jigami

From the  National Institute of Bioscience and Human Technology, Tsukuba, Ibaraki 305, Japan and the ^§ Asahi Chemical Industry Co., Ltd., Fuji, Shizuoka 416, Japan

In yeast Saccharomyces cerevisiae the N-linked sugar chain is modified at different positions by the addition of mannosylphosphate. The mnn6 mutant is deficient in the mannosylphosphate transferase activity toward mannotetraose (Karson, E. M., and Ballou, C. E. (1978) J. Biol. Chem. 253, 6484-6492). We have cloned the MNN6 gene by complementation. It has encoded a 446-amino acid polypeptide with the characteristics of type II membrane protein. The deduced Mnn6p showed a significant similarity to Kre2p/Mnt1p, a Golgi -1,2-mannosyltransferase involved in O-glycosylation. The null mutant of MNN6 showed a normal cell growth, less binding to Alcian blue, hypersensitivity to Calcoflour White and hygromycin B, and diminished mannosylphosphate transferase activity toward the endoplasmic reticulum core oligosaccharide acceptors (Man8GlcNAc2-PA and Man5GlcNAc2-PA) in vitro, suggesting the involvement of the MNN6 gene in the endoplasmic reticulum core oligosaccharide phosphorylation. However, no differences were observed in N-linked mannoprotein oligosaccharides between och1 mnn1 cells and och1mnn1mnn6 cells, indicating the existence of redundant genes required for the core oligosaccharide phosphorylation. Based on a dramatic decrease in polymannose outer chain phosphorylation by MNN6 gene disruption and a determination of the mannosylphosphorylation site in the acceptor, it is postulated that the MNN6 gene may be a structural gene encoding a mannosylphosphate transferase, which recognizes any oligosaccharides with at least one -1,2-linked mannobiose unit.

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