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J. Biol. Chem., Vol. 280, Issue 10, 9489-9497, March 11, 2005

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PIG-V Involved in Transferring the Second Mannose in Glycosylphosphatidylinositol^*

Ji Young Kang, Yeongjin Hong||, Hisashi Ashida, Nobue Shishioh, Yoshiko Murakami, Yasu S. Morita, Yusuke Maeda, and Taroh Kinoshita¶

From the Department of Immunoregulation, Research Institute for Microbial Diseases, 3-1 Yamada-oka, Suita, Osaka 565-0871, Japan and the Department of Microbiology, Genomic Research Center for Enteropathogenic Bacteria, Chonnam National University Medical School, Gwangju 501-746, South Korea

Glycosylphosphatidylinositol (GPI) is a glycolipid that anchors many proteins to the eukaryotic cell surface. The biosynthetic pathway of GPI is mediated by sequential additions of sugars and other components to phosphatidylinositol. Four mannoses in the GPI are transferred from dolichol-phosphate-mannose (Dol-P-Man) and are linked through different glycosidic linkages. Therefore, four Dol-P-Man-dependent mannosyltransferases, GPI-MT-I, -MT-II, -MT-III, and -MT-IV for the first, second, third, and fourth mannoses, respectively, are required for generation of GPI. GPI-MT-I (PIG-M), GPI-MT-III (PIG-B), and GPI-MT-IV (SMP3) were previously reported, but GPI-MT-II remains to be identified. Here we report the cloning of PIG-V involved in transferring the second mannose in the GPI anchor. Human PIG-V encodes a 493-amino acid, endoplasmic reticulum (ER) resident protein with eight putative transmembrane regions. Saccharomyces cerevisiae protein encoded in open reading frame YBR004c, which we termed GPI18, has 25% amino acid identity to human PIG-V. Viability of the yeast gpi18 deletion mutant was restored by human PIG-V cDNA. PIG-V has two functionally important conserved regions facing the ER lumen. Taken together, we suggest that PIG-V is the second mannosyltransferase in GPI anchor biosynthesis.

Received for publication, December 9, 2004 , and in revised form, December 27, 2004.

The nucleotide sequence(s) reported in this paper has been submitted to the GenBank^TM/EBI Data Bank with accession number(s) AB196341.

^* This work was supported in part by grants from the Ministry of Education, Science, Sports, Culture and Technology of Japan. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

^|| Supported in part by a grant from the Ministry of Health and Welfare of the Republic of Korea (01-PJ10-PG6-01GM00-0002).

^¶ To whom correspondence should be addressed. Tel.: 81-6-6879-8328; Fax: 81-6-6875-5233; E-mail tkinoshi{at}biken.osaka-u.ac.jp.

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