Direct interaction and determination of binding domains among peroxisomal import factors in Arabidopsis thaliana
Direct interaction and determination of binding domains among peroxisomal import factors in Arabidopsis thaliana
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DOI：
10.1093/pcp/pcf057
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发表时间：
2002-04-01
影响因子：
4.9
通讯作者：
Nishimura, M
中科院分区：
文献类型：
--
作者：
Nito, K;Hayashi, M;Nishimura, M
我们分析了人Pex14p、Pex5p和Pex7p的拟南芥同源基因的作用，这些Pex14p、Pex5p和Pex7p是过氧体蛋白输入机制的中心组件。免疫印迹分析表明，AtPex14p和AtPex5p存在于拟南芥的大部分器官中，这表明这些因子在植物过氧化物体的主要蛋白质输入途径中发挥作用。双杂交分析表明，AtPex14p与AtPex5p相互作用，但不与AtPex7p相互作用。此外，AtPex7p与AtPex5p结合，表明在拟南芥中，pts2途径依赖于PTS1途径。进一步分析表明，AtPex5p的K-231-(450)D和M-1-V-230的9个WXXXF/Y重复序列分别与AtPex14p的I-58-L-65和R-78-R-97氨基酸以及AtPex7p的C-末端氨基酸Y-266-S-317结合。由于AtPex5p与AtPex14p和AtPex7p的结合域不重叠，AtPex14p、AtPex5p和AtPex7p可能形成它们的复合体，在过氧化体蛋白输入中协同发挥作用。
We analyzed the role of Arabidopsis orthologues of human Pex14p, Pex5p and Pex7p that are central components of peroxisomal protein import machinery. Immunoblot analysis showed that AtPex14p and AtPex5p were present in most organs In Arabidopsis, suggesting that these factors play a role in the main protein import pathways for plant peroxisomes. Two-hybrid analysis showed that AtPex14p interacted with AtPex5p, but not with AtPex7p. In addition, AtPex7p was bound to AtPex5p, indicating that the PTS2 pathway depends on the PTS1 pathway in Arabidopsis. Further analysis showed that the nine WXXXF/Y repeats in the amino acids K-231-(450) D and M-1-V-230 of AtPex5p are bound to two N-terminal domains, amino acids I-58-L-65 and R-78-R-97 of AtPex14p and the C-terminal amino acids Y-266-S-317 of AtPex7p, respectively. Since the binding domains of AtPex5p to AtPex14p and AtPex7p do not overlap, AtPex14p, AtPex5p and AtPex7p might form their complex and function cooperatively in peroxisomal protein import.