Phosphatidylinositol 5-phosphate biosynthesis is linked to PIKfyve and is involved in osmotic response pathway in mammalian cells

被引:101
作者
Sbrissa, D [1 ]
Ikonomov, OC [1 ]
Deeb, R [1 ]
Shisheva, A [1 ]
机构
[1] Wayne State Univ, Sch Med, Dept Physiol, Detroit, MI 48201 USA
关键词
D O I
10.1074/jbc.M207576200
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
The cellular functions, regulation and enzymology of phosphatidylinositol (PtdIns) 5-P, the newest addition to the family of phosphoinositides (PI), are still elusive. Whereas a kinase that uses PtdIns-5-P as an intracellular substrate has been assigned, a kinase that produces it remained to be identified. Here we report that PIKfyve, the enzyme found to synthesize PtdIns-5-P in vitro and PtdIns-3,5-P-2 in vitro and in vivo, is responsible for PtdIns-5-P production in a cellular context. Evidence is based on examination of two groups of cell types by two independent approaches. First, [P-32]orthophosphate-labeled cells (Sf9, 3T3-L1 fibroblasts, and 3T3-L1 adipocytes) that show a high pressure liquid chromatography (HPLC)-detectable peak of the PtdIns-5-P head group at basal conditions demonstrated a 20-50% increase in radioactive PtdIns-5-P amounts upon expression of PIK-fyve(WT). Second, cell types (HEK293), in which the basal levels of radioactive PtdIns-5-P were undetectable by HPLC head group analysis, demonstrated higher in vitro type II PIP kinase-directed conversion of the endogenous PtdIns-5-P pool into PtdIns-4,5-P-2, when induced to express PIKfyveWT. Conversely, a decrease by 60% in the conversion of PtdIns-5-P to PtdIns-4,5-P-2 was associated with induced expression of the dominant-negative kinase-deficient PIKfyve(K1831E) mutant in HEK293 cells. When 3T3-L1 fibroblasts and 3T3-L1 adipocytes were subjected to osmotic shock, levels of PtdIns-5-P measured by both approaches were found to decrease profoundly upon a hypo-osmotic stimulus. Together, these results identify PIKfyve as an enzyme responsible for PtdIns-5-P biosynthesis and indicate a role for PtdIns-5-P in osmotic response pathways in mammalian cells.
引用
收藏
页码:47276 / 47284
页数:9
相关论文
共 39 条
[1]   Phosphatidylinositol phosphate kinases, a multifaceted family of signaling enzymes [J].
Anderson, RA ;
Boronenkov, IV ;
Doughman, SD ;
Kunz, J ;
Loijens, JC .
JOURNAL OF BIOLOGICAL CHEMISTRY, 1999, 274 (15) :9907-9910
[2]   Osmotic stress-induced increase of phosphatidylinositol 3,5-bisphosphate requires Vac14p, an activator of the lipid kinase Fab1p [J].
Bonangelino, CJ ;
Nau, JJ ;
Duex, JE ;
Brinkman, M ;
Wurmser, AE ;
Gary, JD ;
Emr, SD ;
Weisman, LS .
JOURNAL OF CELL BIOLOGY, 2002, 156 (06) :1015-1028
[3]   A novel interaction between the juxtamembrane region of the p55 tumor necrosis factor receptor and phosphatidylinositol-4-phosphate 5-kinase [J].
Castellino, AM ;
Parker, GJ ;
Boronenkov, IV ;
Anderson, RA ;
Chao, MV .
JOURNAL OF BIOLOGICAL CHEMISTRY, 1997, 272 (09) :5861-5870
[4]  
Chen D, 1999, MOL CELL BIOL, V19, P4684
[5]   Osmotic shock stimulates GLUT4 translocation in 3T3L1 adipocytes by a novel tyrosine kinase pathways [J].
Chen, D ;
Elmendorf, JS ;
Olson, AL ;
Li, X ;
Earp, S ;
Pessin, JE .
JOURNAL OF BIOLOGICAL CHEMISTRY, 1997, 272 (43) :27401-27410
[6]   Inositol lipids are regulated during cell cycle progression in the nuclei of murine erythroleukaemia cells [J].
Clarke, JH ;
Letcher, AJ ;
D'Santos, CS ;
Halstead, JR ;
Irvine, RF ;
Divecha, N .
BIOCHEMICAL JOURNAL, 2001, 357 (03) :905-910
[7]   Phosphoinositides in membrane traffic [J].
Corvera, S ;
D'Arrigo, A ;
Stenmark, H .
CURRENT OPINION IN CELL BIOLOGY, 1999, 11 (04) :460-465
[8]   Direct targets of phosphoinositide 3-kinase products in membrane traffic and signal transduction [J].
Corvera, S ;
Czech, MP .
TRENDS IN CELL BIOLOGY, 1998, 8 (11) :442-446
[9]   Rapid accumulation of phosphatidylinositol 4,5-bisphosphate and inositol 1,4,5-trisphosphate correlates with calcium mobilization in salt-stressed Arabidopsis [J].
DeWald, DB ;
Torabinejad, J ;
Jones, CA ;
Shope, JC ;
Cangelosi, AR ;
Thompson, JE ;
Prestwich, GD ;
Hama, H .
PLANT PHYSIOLOGY, 2001, 126 (02) :759-769
[10]   Vac14 controls PtdIns(3,5)P2 synthesis and Fab1-dependent protein trafficking to the multivesicular body [J].
Dove, SK ;
McEwen, RK ;
Mayes, A ;
Hughes, DC ;
Beggs, JD ;
Michell, RH .
CURRENT BIOLOGY, 2002, 12 (11) :885-893