PtdIns4P recognition by Vps74/GOLPH3 links PtdIns 4-kinase signaling to retrograde Golgi trafficking

Abstract

Retention of Golgi-resident integral membrane proteins depends on the supply of PI(4)P.

An Oncogenic Protein Golgi Phosphoprotein 3 Up-regulates Cell Migration via Sialylation

“…The cDNA of HA-tagged GOLPH3 (a generous gift from Dr. Lynda Chin, Institute for Applied Cancer Science, University of Texas MD Anderson Cancer Center) (21) and HA-tagged, shRNA resistance GOLPH3 were cloned by standard PCR protocols, into pENTR/D-TOPO (Invitrogen). The overlap extension PCRs were used to construct GOLPH3 mutants, which lacks a tetramer formation (⌬190 -201) (28) or the binding to PI4P (R171A/R174A and W81A/R90A) (29). The cDNAs of human ␣2,3-sialyltransferase-IV (ST3GAL4), ST6GAL1, and ␤1,4-galactosyltransferase I (␤4GALT1) (kindly provided by Dr. H. Narimatsu from National Institute of Advanced Industrial Science and Technology, Japan) were inserted into pENTR 1A tagged with 3ϫFLAG at the C terminus using the in-fusion method (Takara Bio).…”

“…GOLPH3 Was Specifically Associated with SialyltransferasesIn yeast cells, Vps74p, which is a counterpart of GOLPH3, was associated with mannosyltransferase enzyme (26), coatomer (45), phosphatidylinositol 4-phosphate (PI4P) (29), and regulated N-glycan synthesis. Very recently, GCNT1 enzyme was reported to be one of the putative binding partners for GOLPH3 to retain its Golgi localization (27).…”

“…These results suggest that GOLPH3 is able to bind to a specific region within the cytoplasmic domains of sialyltransferases. In fact, GOLPH3 recognized PI4P via a positively charged binding pocket on the hydrophobic face of the protein (22,29) for localization in the trans-Golgi. Vps74p oligomer is reported to be required for Golgi localization of glycosyltransferases in yeast cells (16).…”

“…Vps74p oligomer is reported to be required for Golgi localization of glycosyltransferases in yeast cells (16). Therefore, we next sought to determine whether a lack of PI4P binding ability (R171A/R174A or W81A/R90A) (29) or the disruption of its oligomerization (⌬190 -201) (16) rescued the aberrant sialylation observed in GOLPH3 knockdown cells. As expected, these GOLPH3 mutants could not rescue the sialylation levels of ␤1 integrin, suggesting the GOLPH3/sialyltransferase/PI4P participated in the regulation of ␣2,6-sialylations in a coordinated fashion (Fig.…”

Lentivirus Mediated GOLPH3 shRNA Inhibits Growth and Metastasis of Esophageal Squamous Cancer

“…GOLPH3 was initially detected as a phosphorylated protein localized to the Golgi apparatus (Dippold et al, 2009;Scott and Chin 2010). Recently, the cell biological, biochemical and functional analyses have shown that GOLPH3 can increase cell transformation and tumor growth by enhancing the activity of mTORC (Dippold et al, 2009;Wood et al, 2009;Scott and Chin 2010;Graham and Burd, 2011). Moreover, the preliminary First Affiliated Hospital, Henan University of Science and Technology,Luoyang,Henan,China *For correspondence: wangqiang_henan@163.com Qiang Wang* , Xian Wang, Can-Bin Zhang researches shown that GOLPH3 is relative with poor prognosis of some tumors, including gastric cancer (Hu et al, 2013) and esophageal squamous cell carcinoma (Wang et al, 2012).…”