Identification of strain-specific variants of mouse Adamts13 gene encoding von Willebrand factor-cleaving protease
Identification of strain-specific variants of mouse Adamts13 gene encoding von Willebrand factor-cleaving protease
复制标题
DOI：
10.1074/jbc.m314184200
复制
发表时间：
2004-07-16
影响因子：
4.8
通讯作者：
Miyata, T
中科院分区：
文献类型：
--
作者：
Banno, F;Kaminaka, K;Miyata, T
人ADAMTS 13是最近发现的一种编码血管性血友病因子裂解蛋白酶的基因。这种酶的先天性和后天性缺陷均可引起血栓性血小板减少性紫癜。hADAMTS 13由1，427个氨基酸残基组成，并且由包括血小板反应蛋白1型基序和CUB结构域的多个结构域组成。为了分析这些结构域在体内的功能作用，我们确定了小鼠直系同源物mADAMTS 13的cDNA序列。出乎意料的是，两种形式的小鼠Adamts 13基因被分离出来，其不同之处在于插入了包括提前终止密码子的脑池内A颗粒（IAP）反转录转座子。在BALB/c、C3 H/He、C57 BL/6和DBA/2株中发现IAP插入，而在129/Sv株中未发现IAP插入。远交ICR品系具有IAP-自由或IAP-插入等位基因或两者。不含IAP的Adamts 13编码mADAMTS 13 L，这是一种具有与hADAMTS 13相同结构域的1，426个氨基酸残基的蛋白质。相反，IAP插入的Adamts 13编码C末端截短的酶mADAMTS 13 S，其仅由1，037个氨基酸残基组成，并且缺乏C末端的两个1型血小板反应蛋白基序和两个CUB结构域。逆转录-PCR和北方印迹分析也证实了菌株特异性。重组mADAMTS 13 L和mADAMTS 13 S在体外均表现出血管性血友病因子切割活性。小鼠ADAMTS 13的自然变异应允许测定其C-末端结构域在体内迄今未知的功能。
Human ADAMTS13 was recently identified as a gene encoding von Willebrand factor-cleaving protease, hADAMTS13. Both congenital and acquired defects in this enzyme can cause thrombotic thrombocytopenic purpura. hADAMTS13 consists of 1,427 amino acid residues and is composed of multiple structural domains including thrombospondin type 1 motifs and CUB domains. To analyze the functional roles of these domains in vivo, we determined the cDNA sequence of the mouse ortholog, mADAMTS13. Unexpectedly, two forms of the mouse Adamts13 gene were isolated that differed in the insertion of an intracisternal A particle (IAP) retrotransposon including a premature stop codon. The IAP insertion was found in BALB/c, C3H/He, C57BL/6, and DBA/2 strains but not in the 129/Sv strain. The outbred ICR strain had either the IAP-free or IAP-inserted allele or both. IAP-free Adamts13 encoded mADAMTS13L, a protein of 1,426 amino acid residues with the same domain organization as hADAMTS13. In contrast, IAP-inserted Adamts13 encoded a C-terminally truncated enzyme, mADAMTS13S, that is comprised of only 1,037 amino acid residues and lacking the C-terminal two thrombospondin type 1 motifs and two CUB domains. Strain specificity was also confirmed by reverse transcription-PCR and Northern blot analyses. Both recombinant mADAMTS13L and mADAMTS13S exhibited von Willebrand factor cleaving activities in vitro. The natural variation in mouse ADAMTS13 should allow for the determination of hitherto unknown functions of its C-terminal domains in vivo.