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首页> 外文期刊>Proceedings of the National Academy of Sciences of the United States of America >Molecular cloning of TRPC3a, an N-terminally extended, store-operated variant of the human C3 transient receptor potential channel
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Molecular cloning of TRPC3a, an N-terminally extended, store-operated variant of the human C3 transient receptor potential channel

机译：TRPC3a的分子克隆，它是人C3瞬时受体电位通道的N端延伸的，由存储操纵的变异体

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摘要

AK032317 is the GenBank accession no. of a full-length RIKEN mouse cDNA. It encodes a putative variant of the C3-type TRPC (transient receptor potential channel) that differs from the previously cloned murine TRPC3 cDNA in that it has a 5′ extension stemming from inclusion of an additional exon (exon 0). The extended cDNA adds 62 aa to the sequence of the murine TRPC3. Here, we report the cloning of a cDNA encoding the human homologue of this extended TRPC3 having a highly homologous 73-aa N-terminal extension, referred to as hTRPC3a. A query of the GenBank genomic database predicts the existence of a similar gene product also in rats. Transient expression of the longer TRPC3a in human embryonic kidney (HEK) cells showed that it mediates Ca~(2+) entry in response to stimulation of the Gq-phospholipase C β pathway, which is similar to that mediated by the shorter hTRPC3. However, after isolation of HEK cells expressing hTRPC3 in stable form, TRPC3a gave rise to Ca~(2+)-entry channels that are not only activated by the Gq-phospholipase C β pathway (receptor-activated Ca entry) but also by thapsigargin triggered store depletion. In conjunction with findings from our and other laboratories that TRPC1, TRPC2, TRPC4, TRPC5, and TRPC7, can each mediate store-depletion-activated Ca~(2+) entry in mammalian cells, our findings with hTRC3a support our previous proposal that TRPCs form capacitative Ca-entry channels.
机译：AK032317是GenBank的登录号。 RIKEN小鼠全长cDNA的构建。它编码一个C3型TRPC（瞬时受体电位通道）的推定变体，该变体与先前克隆的鼠TRPC3 cDNA不同，因为它具有5'延伸，这是由于包含了一个额外的外显子（外显子0）。延伸的cDNA向鼠类TRPC3的序列增加了62个氨基酸。在这里，我们报告了一个cDNA的克隆，该cDNA编码具有高度同源的73-aa N端延伸（称为hTRPC3a）的这种延伸的TRPC3的人类同源物。 GenBank基因组数据库的查询可预测大鼠中也存在相似的基因产物。较长的TRPC3a在人胚肾脏（HEK）细胞中的瞬时表达表明，它响应Gq-磷脂酶Cβ途径的刺激而介导Ca〜（2+）进入，这与较短的hTRPC3介导的相似。然而，在分离以稳定形式表达hTRPC3的HEK细胞后，TRPC3a产生了Ca〜（2+）进入通道，该通道不仅被Gq-磷脂酶Cβ途径（受体激活的Ca进入）激活，而且也被毒胡萝卜素激活。触发商店枯竭。结合我们和其他实验室的发现，即TRPC1，TRPC2，TRPC4，TRPC5和TRPC7均可以介导哺乳动物细胞中由存储耗尽激活的Ca〜（2+）进入，我们在hTRC3a中的发现支持了我们先前关于TRPC的提议形成电容性进入通道。

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