TWISTED DWARF1, a unique plasma membrane-anchored immunophilin-like protein, interacts with Arabidopsis multidrug resistance-like transporters AtPGP1 and AtPGP19
TWISTED DWARF1, a unique plasma membrane-anchored immunophilin-like protein, interacts with Arabidopsis multidrug resistance-like transporters AtPGP1 and AtPGP19
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DOI：
10.1091/mbc.e02-10-0698
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发表时间：
2003-10-01
影响因子：
3.3
通讯作者：
Schulz, B
中科院分区：
文献类型：
--
作者：
Geisler, M;Kolukisaoglu, HÜ;Schulz, B
拟南芥FKBP样亲免素TWISTED DWARF 1（TWDI）基因的零突变导致多效性表型，其特征在于细胞伸长减少和所有植物器官的定向生长。异源表达的TWD 1不表现出顺式-反式-肽基脯氨酰异构酶（PPI酶）活性，并且不与酵母FKBP 12突变体互补，这表明TWD 1通过蛋白质-蛋白质相互作用间接起作用。酵母双杂交蛋白质相互作用筛选与TWD 1确定的cDNA序列编码的C-末端结构域的拟南芥多药耐药样ABC转运蛋白AtPGP 1。在体外验证了这种相互作用。蛋白质相互作用结构域的映射显示，AtPGP 1令人惊讶地结合到N-末端的TWD 1窝藏的顺式-反式肽基脯氨酰异构酶样结构域，而不是tetratrico肽重复结构域，这已被证明介导蛋白质-蛋白质相互作用。与所有其他FKBP不同，TWD 1被证明是一种整合的膜蛋白，与其相互作用的伴侣AtPGP 1在质膜上共定位。TWD 1还与AtPGP 19（AtMDR 1）相互作用，AtPGP 19是AtPGP 1最接近的同源物。单基因突变twd 1 -1和双atpgp 1 -1/atpgp 19 -1（atmdr 1 -1）突变体表现出相似的表型，包括上位性生长，花序大小减少，极性生长素运输减少，这表明一个功能TWD 1-AtPGP 1/AtPGP 19复合体是必要的适当的植物发育。
Null-mutations of the Arabidopsis FKBP-like immunophilin TWISTED DWARF1 (TWDI) gene cause a pleiotropic phenotype characterized by reduction of cell elongation and disorientated growth of all plant organs. Heterologously expressed TWD1 does not exhibit cis-trans-peptidylprolyl isomerase (PPIase) activity and does not complement yeast FKBP12 mutants, suggesting that TWD1 acts indirectly via protein-protein interaction. Yeast two-hybrid protein interaction screens with TWD1 identified cDNA sequences that encode the C-terminal domain of Arabidopsis multidrug-resistance-like ABC transporter AtPGP1. This interaction was verified in vitro. Mapping of protein interaction domains shows that AtPGP1 surprisingly binds to the N-terminus of TWD1 harboring the cis-trans peptidyl-prolyl isomerase-like domain and not to the tetratrico-peptide repeat domain, which has been shown to mediate protein-protein interaction. Unlike all other FKBPs, TWD1 is shown to be an integral membrane protein that colocalizes with its interacting partner AtPGP1 on the plasma membrane. TWD1 also interacts with AtPGP19 (AtMDR1), the closest homologue of AtPGP1. The single gene mutation twd1-1 and double atpgp1-1/atpgp19-1 (atmdr1-1) mutants exhibit similar phenotypes including epinastic growth, reduced inflorescence size, and reduced polar auxin transport, suggesting that a functional TWD1-AtPGP1/AtPGP19 complex is required for proper plant development.