TY - JOUR
T1 - Comparative Haploid Genetic Screens Reveal Divergent Pathways in the Biogenesis and Trafficking of Glycophosphatidylinositol-Anchored Proteins
AU - Davis, Eric M.
AU - Kim, Jihye
AU - Menasche, Bridget L.
AU - Sheppard, Jacob
AU - Liu, Xuedong
AU - Tan, Aik Choon
AU - Shen, Jingshi
N1 - Funding Information:
We thank Dr. Thijn Brummelkamp for providing HAP1 cells and gene-trap plasmids, Dr. Ramanujan Hegde for providing PrP-related plasmids, Yan Ouyang for technical assistance, Yuming Han for assistance with flow cytometry, Dr. Jolien Tyler for assistance with confocal imaging, and Dr. Joe Heimiller for discussions on bioinformatics. We are grateful to Drs. Peter Orlean, Erik Snapp, and Thomas Rutkowski for insightful comments on the manuscript. We thank the Functional Genomics Facility at the University of Colorado Cancer Center for providing the expression plasmids of SEC62 and SEC63. This work was supported by the NIH grant GM102217 (J.S.), a Pew Scholar Award (J.S.), and a seed grant from the Cancer League of Colorado (A.-C.T.).
Publisher Copyright:
© 2015 The Authors.
PY - 2015/6/23
Y1 - 2015/6/23
N2 - Glycophosphatidylinositol-anchored proteins (GPI-APs) play essential roles in physiology, but their biogenesis and trafficking have not been systematically characterized. Here, we took advantage of the recently available haploid genetics approach to dissect GPI-AP pathways in human cells using prion protein (PrP) and CD59 as model molecules. Our screens recovered a large number of common and unexpectedly specialized factors in the GPI-AP pathways. PIGN, PGAP2, and PIGF, which encode GPI anchor-modifying enzymes, were selectively isolated in the CD59 screen, suggesting that GPI anchor composition significantly influences the biogenesis of GPI-APs in a substrate-dependent manner. SEC62 and SEC63, which encode components of the ER-targeting machinery, were selectively recovered in the PrP screen, indicating that they do not constitute a universal route for the biogenesis of mammalian GPI-APs. Together, these comparative haploid genetic screens demonstrate that, despite their similarity in overall architecture and subcellular localization, GPI-APs follow markedly distinct biosynthetic and trafficking pathways. Using comparative haploid genetic screens in human cells, Davis et al. find that GPI-anchored proteins follow markedly distinct biosynthetic and trafficking pathways in spite of their similarity in overall architecture and subcellular localization.
AB - Glycophosphatidylinositol-anchored proteins (GPI-APs) play essential roles in physiology, but their biogenesis and trafficking have not been systematically characterized. Here, we took advantage of the recently available haploid genetics approach to dissect GPI-AP pathways in human cells using prion protein (PrP) and CD59 as model molecules. Our screens recovered a large number of common and unexpectedly specialized factors in the GPI-AP pathways. PIGN, PGAP2, and PIGF, which encode GPI anchor-modifying enzymes, were selectively isolated in the CD59 screen, suggesting that GPI anchor composition significantly influences the biogenesis of GPI-APs in a substrate-dependent manner. SEC62 and SEC63, which encode components of the ER-targeting machinery, were selectively recovered in the PrP screen, indicating that they do not constitute a universal route for the biogenesis of mammalian GPI-APs. Together, these comparative haploid genetic screens demonstrate that, despite their similarity in overall architecture and subcellular localization, GPI-APs follow markedly distinct biosynthetic and trafficking pathways. Using comparative haploid genetic screens in human cells, Davis et al. find that GPI-anchored proteins follow markedly distinct biosynthetic and trafficking pathways in spite of their similarity in overall architecture and subcellular localization.
UR - http://www.scopus.com/inward/record.url?scp=84937634471&partnerID=8YFLogxK
U2 - 10.1016/j.celrep.2015.05.026
DO - 10.1016/j.celrep.2015.05.026
M3 - Article
C2 - 26074080
AN - SCOPUS:84937634471
SN - 2211-1247
VL - 11
SP - 1727
EP - 1736
JO - Cell Reports
JF - Cell Reports
IS - 11
ER -