PPM1D430, a Novel Alternative Splicing Variant of the Human PPM1D, can Dephosphorylate p53 and Exhibits Specific Tissue Expression
PPM1D430, a Novel Alternative Splicing Variant of the Human PPM1D, can Dephosphorylate p53 and Exhibits Specific Tissue Expression
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DOI：
10.1093/jb/mvn135
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发表时间：
2009-01-01
影响因子：
2.7
通讯作者：
Sakaguchi, Kazuyasu
中科院分区：
文献类型：
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作者：
Chuman, Yoshiro;Kurihashi, Wataru;Sakaguchi, Kazuyasu
PPM1D是一种PPM1类型的蛋白磷酸酶，在DNA损伤时被诱导。PPM1D缺陷小鼠在精子发生和淋巴细胞功能方面存在缺陷，但这些表型背后的机制尚不清楚。在我们目前的研究中，我们在mRNA和蛋白质水平上鉴定和鉴定了人PPM1D的一个选择性剪接变异体(标记为PPM1D430)。PPM1D430由已知PPM1D蛋白(PPM1D605)的常见420个残基组成，并含有PPM1D430特异的10个氨基酸。半定量逆转录聚合酶链式反应(RTPCR)分析表明，PPM1D430mRNA也以P53依赖的方式在遗传毒性应激中被诱导。体外磷酸酶分析和PPM1D430特异的RNA干扰分析进一步表明，PPM1D430在体外和体内都能使人P53的Ser15去磷酸化。另一方面，通过RT-PCR对该基因的表达谱分析表明，PPM1D430仅在睾丸和白细胞中表达，而PPM1D605普遍存在。此外，在某些条件下，PPM1D430与PPM1D605相比，表现出不同的亚细胞定位模式和蛋白质稳定性。因此，我们目前的发现表明，PPM1D430可能在免疫反应和/或精子发生中发挥特定的功能。
PPM1D is a PPM1 type protein phosphatase and is induced in response to DNA damage. PPM1D-deficient mice show defects in spermatogenesis and lymphoid cell functions but the mechanisms underlying these phenotypes remain unknown. In our current study, we identify and characterize an alternative splicing variant (denoted PPM1D430) of human PPM1D at both the mRNA and protein level. PPM1D430 comprises the common 420 residues of the known PPM1D protein (PPM1D605) and contains a stretch of PPM1D430-specific 10 amino acids. Semi-quantitative reverse transcriptionpolymerase chain reaction (RTPCR) analysis revealed that PPM1D430 mRNA is also induced in response to the genotoxic stress in a p53-dependent manner. In vitro phosphatase analysis and PPM1D430-specific RNA interference analysis further indicated that PPM1D430 can dephosphorylate Ser15 of human p53 both in vitro and in vivo. On the other hand, expression profiling of this gene by RTPCR analysis of a human tissue cDNA panel revealed that PPM1D430 is expressed exclusively in testes and in leucocytes whereas PPM1D605 is ubiquitous. In addition, PPM1D430 shows a different subcellular localization pattern and protein stability when compared with PPM1D605 under some conditions. Our current findings thus suggest that PPM1D430 may exert specific functions in immune response and/or spermatogenesis.