The normal parkin sequence.
The normal parkin sequence.
复制标题
正常的停车顺序。
DOI：
10.1002/mds.24050
复制
发表时间：
2012
期刊：
影响因子：
--
通讯作者：
Feng,Jian
中科院分区：
文献类型：
--
作者：
Ren,Yong;Liu,Xiaojun;Lesage,Suzanne;Cai,Miao;Pu,Jiali;Zhang,Baorong;Brice,Alexis;Feng,Jian
The recruitment of parkin to mitochondria in response to mitochondrial membrane depolarization1 is an important aspect of parkin biology that has attracted extensive research2. We found that parkin with the sequence (AB009973. 1) reported in the original discovery of the gene3 was not recruited to mitochondria following CCCP treatment, while parkin with the current Genbank Reference Sequence (NM_004562. 2) was robustly recruited to mitochondria under the same condition. The only difference between the two sequences is nucleotide 768, which is C in AB009973. 1 (encoding for proline at amino acid position 223) and T in NM_004562. 2 (for serine at 223). As shown in Fig. 1, Hela cells were transfected with FLAG-tagged parkin (with either P or S at amino acid 223). Cells were treated without or with CCCP (10 μM) for 3 hr and immunostained for FLAG and Tom20. CCCP induced a strong mitochondrial recruitment of FLAG-parkin (223S), but no significant recruitment of FLAG-parkin (223P). This was confirmed using GFP-parkin with either P or S at position 223 (Fig. 1C). Similar results were also obtained in COS7 or HEK293 cells using FLAG-, Myc-, HA-or GFP-tagged human parkin with S or P at amino acid 223 (data not shown).To test which version of parkin sequence is correct, we analyzed genomic DNA of 2102 individuals from Europe, China and the United States. None of them had C at this position; all were T. Briefly, genomic DNA from 231 normal subjects and 294 Parkinson’s disease patients in China, as well as genomic DNA from 19 normal subjects and 18 Parkinson’s disease patients in the United States were amplified by PCR using two primers in the introns flanking exon 6 of parkin (CTTGTCCAAAGAGATTGTTTACTGTGG and GGCTCGTGTGGCAGAACAATATTGGG). The PCR product (318bp) should contain a single BsrI site (CCAGT) if AB009973. 1 is correct. None of these samples could be cut by
