Stereo-specific substrate recognition by phosphatidylinositol phosphate kinases is swapped by changing a single amino acid residue

被引:63
作者
Kunz, J [1 ]
Fuelling, A [1 ]
Kolbe, L [1 ]
Anderson, RA [1 ]
机构
[1] Univ Wisconsin, Med Sch, Dept Pharmacol, Madison, WI 53706 USA
关键词
D O I
10.1074/jbc.M110775200
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
Type I and type II phosphatidylinositol phosphate (PIP) kinases generate the lipid second messenger phosphatidylinositol (PtdIns) 4,5-bisphosphate and thus play fundamental roles in the regulation of many cellular processes. Although the two kinase families are highly homologous, they phosphorylate distinct substrates and are functionally non-redundant. Type I PIP kinases phosphorylate PtdIns 4-phosphate at the D-5 hydroxyl group and are consequently PtdIns 4-phosphate 5-kinases. By contrast, type II PIP kinases are PtdIns 5-phosphate 4-kinases that phosphorylate PtdIns 5-phosphate at the D-4 position. Type I PIP kinases, in addition, also phosphorylate other phosphoinositides in vitro and in vivo and thus have the potential to generate multiple lipid second messengers. To understand how these enzymes differentiate between stereoisomeric substrates, we used a site-directed mutagenesis approach. We show that a single amino acid substitution in the activation loop, A381E in IIbeta and the corresponding mutation E362A in Ibeta, is sufficient to swap substrate specificity between these PIP kinases. In addition to its role in substrate specificity, the type I activation loop is also key in subcellular targeting. The Ibeta(E362A) mutant and other mutants with reduced PtdIns 4-phosphate binding affinity were largely cytosolic when expressed in mammalian cells in contrast to wild-type Ibeta which targets to the plasma membrane. These results clearly establish the role of the activation loop in determining both signaling specificity and plasma membrane targeting of type I PIP kinases.
引用
收藏
页码:5611 / 5619
页数:9
相关论文
共 46 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]   Coupled inositide phosphorylation and phospholipase D activation initiates clathrin-coat assembly on lysosomes [J].
Arneson, LS ;
Kunz, J ;
Anderson, RA ;
Traub, LM .
JOURNAL OF BIOLOGICAL CHEMISTRY, 1999, 274 (25) :17794-17805
[3]   Bifurcation of lipid and protein kinase signals of PI3Kγ to the protein kinases PKB and MAPK [J].
Bondeva, T ;
Pirola, L ;
Bulgarelli-Leva, G ;
Rubio, I ;
Wetzker, R ;
Wymann, MP .
SCIENCE, 1998, 282 (5387) :293-296
[4]   Phosphoinositide signaling pathways in nuclei are associated with nuclear speckles containing pre-mRNA processing factors [J].
Boronenkov, IV ;
Loijens, JC ;
Umeda, M ;
Anderson, RA .
MOLECULAR BIOLOGY OF THE CELL, 1998, 9 (12) :3547-3560
[5]   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
[6]   Nuclear targeting of the β isoform of Type II phosphatidylinositol phosphate kinase (phosphatidylinositol 5-phosphate 4-kinase) by its α-helix 7 [J].
Ciruela, A ;
Hinchliffe, KA ;
Divecha, N ;
Irvine, RF .
BIOCHEMICAL JOURNAL, 2000, 346 (pt 3) :587-591
[7]   PIP2 and PIP3: Complex roles at the cell surface [J].
Czech, MP .
CELL, 2000, 100 (06) :603-606
[8]   MSS4, a phosphatidylinositol-4-phosphate 5-kinase required for organization of the actin cytoskeleton in Saccharomyces cerevisiae [J].
Desrivières, S ;
Cooke, FT ;
Parker, PJ ;
Hall, MN .
JOURNAL OF BIOLOGICAL CHEMISTRY, 1998, 273 (25) :15787-15793
[9]   Phosphoinositide kinases [J].
Fruman, DA ;
Meyers, RE ;
Cantley, LC .
ANNUAL REVIEW OF BIOCHEMISTRY, 1998, 67 :481-507
[10]   Phosphoinositide binding domains: Embracing 3-phosphate [J].
Fruman, DA ;
Rameh, LE ;
Cantley, LC .
CELL, 1999, 97 (07) :817-820