Investigation of intra-Golgi zone for maturation and transport of GPI-anchored proteins
Glycosylphosphatidylinositols (GPI) are glycolipids that are used as membrane anchors of many cell surface proteins in eukaryotic cells. At least 150 different human proteins are GPI-anchored proteins (GPI-APs). GPI-anchor confers proteins with particular characteristics such as association with lipid microdomains, shedding after cleavage within GPI and apical transport in polarized cells. GPI is essential for life as shown by knockout of a gene involved in GPI biosynthesis that caused early death of mouse embryos. In recent years, taking advantage of whole exome sequencing, individuals who have inherited partial loss-of-function mutations in genes involved in GPI biosynthesis have been found. Clinical symptoms of these individuals indicate roles of GPI-APs in various organs and tissues. Loss-of-function mutations in genes involved in maturation of GPI-APs also cause various phenotypes, indicating that not only amounts but also structure of GPI is critical for human health.
Recent studies demonstrated variations in behaviors of various GPI-APs and possible contributions of side-chain modifications. However, how GPI side-chain modifications are regulated is only partially understood. Because molecular basis of structural variation in GPI side-chains is critical for biology and medicine of GPI-APs, our current study is focused on genes involved in GPI side-chain modifications. In many GPI-APs, N-acetylgalactosamine (GalNAc) is linked to the first mannose in β4 linkage as a side-chain. The GalNAc side-chain can be elongated byβ3 linked galactose (Gal) and further by sialic acid (Figure). Physiological roles of this GalNAc side-chain has been unclear because genes involved have not been identified. Extents of structural variation of the GalNAc side-chains in GPI-APs and tissues are not well understood either. Intra Golgi localization of relevant enzymes is not known.
We recently identified gene encoding Golgi-residentβ4 GalNAc transferase that mediates transfer of GalNAc to the first mannose (Hirata et al, 2018, Nat Commun). We termed the gene PGAP4 for Post GPI Attachment to Proteins 4. We now work on cloning b3Gal transferase that mediates GalNAc side-chain elongation by Gal. A goal in this joint study is to determine where GPI-AP maturation (GPI fatty acid remodeling and side-chain modification) occurs in the Golgi and whether it occurs in a specific zone.
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- Nuclear envelope localization of PIG-B is essential for GPI-anchor synthesis in Drosophila.
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