The highly charged region of plant β-type phosphatidylinositol 4-kinase is involved in membrane targeting and phospholipid binding

被引:17
作者
Lou, Y
Ma, H
Lin, WH
Chu, ZQ
Mueller-Roeber, B
Xu, ZH
Xue, HW
机构
[1] Chinese Acad Sci, Shanghai Inst Biol Chem, Inst Plant Physiol & Ecol, Natl Key Lab Plant Mol Genet, Shanghai 200032, Peoples R China
[2] Max Planck Inst Mol Plant Physiol Plant Mol Physi, Partner Grp, MPIMP, Shanghai 200032, Peoples R China
[3] Univ Potsdam, Inst Biochem & Biol, D-14476 Golm, Germany
关键词
GFP; membrane targeting; PI4K; PPC (Plant PI4Ks Charged)region; signal transduction; yeast;
D O I
10.1007/s11103-005-5548-x
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
In Arabidopsis thaliana and Oryza sativa, two types of PI 4-kinase (PI4Ks) have been isolated and functionally characterized. The alpha-type PI4Ks (similar to 220 kDa) contain a PH domain, which is lacking in beta-type PI4Ks (similar to 120 kDa). beta-Type PI4Ks, exemplified by Arabidopsis AtPI4K beta and rice OsPI4K2, contain a highly charged repetitive segment designated PPC (Plant PI4K Charged) region, which is an unique domain only found in plant beta-type PI4Ks at present. The PPC region has a length of similar to 300 amino acids and harboring 11 (AtPI4K beta) and 14 (OsPI4K2) repeats, respectively, of a 20-aa motif. Studies employing a modified yeast-based "Sequence of Membrane-Targeting Detection'' system demonstrate that the PPC(OsPI4K2) region, as well as the former 8 and latter 6 repetitive motifs within the PPC region, are able to target fusion proteins to the plasma membrane. Further detection on the transiently expressed GFP fusion proteins in onion epidermal cells showed that the PPC(OsPI4K2) region alone, as well as the region containing repetitive motifs 1-8, was able to direct GFP to the plasma membrane, while the regions containing less repetitive motifs, i.e. 6, 4, 2 or single motif(s) led to predominantly intracellular localization. Agrobacterium-mediated transient expression of PPC-GFP fusion protein further confirms the membrane-targeting capacities of PPC region. In addition, the predominant plasma membrane localization of AtPI4Kb was mediated by the PPC region. Recombinant PPC peptide, expressed in E. coli, strongly binds phosphatidic acid, PI and PI4P, but not phosphatidylcholine, PI5P, or PI(4,5) P-2 in vitro, providing insights into potential mechanisms for regulating sub-cellular localization and lipid binding for the plant beta-type PI4Ks.
引用
收藏
页码:729 / 746
页数:18
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