Involvement of TRPV3 and TRPM8 ion channel proteins in induction of mammalian cold-inducible proteins
Involvement of TRPV3 and TRPM8 ion channel proteins in induction of mammalian cold-inducible proteins
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DOI：
10.1016/j.bbrc.2017.11.136
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发表时间：
2018-01-01
影响因子：
3.1
通讯作者：
Nishiyama, Hiroyuki
中科院分区：
文献类型：
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作者：
Fujita, Takanori;Liu, Yu;Nishiyama, Hiroyuki
冷诱导RNA结合蛋白（CIRP）、RNA结合基序蛋白3（RBM 3）和富含丝氨酸和精氨酸的剪接因子5（SRSF 5）是在哺乳动物细胞中响应于适度低温和多种细胞应激而转录上调的RNA结合蛋白。这些冷诱导蛋白（CIP）的诱导依赖于瞬时受体电位（TRP）V4通道蛋白，但似乎不依赖于其离子通道活性。我们在此报告，除了TRPV 4，TRPV 3和TRPM 8是诱导CIP所必需的。我们从TRPV 4基因敲除（KO）小鼠的肺中建立了细胞系，并通过蛋白质印迹分析观察了它们中CIP的诱导。TRPV 4拮抗剂RN 1734抑制野生型小鼠细胞的诱导，但不抑制TRPV 4-KO细胞。TRPV 3通道阻断剂5408271和TRPM 8通道阻断剂AMTB以及针对TRPV 3和TRPM 8的siRNA抑制小鼠TRPV 4-KO细胞和人U-2 OS细胞中的CIP诱导。TRPV 3通道激动剂2-APB诱导CIP表达，但樟脑不诱导。TRPM 8通道激动剂WS-12也没有。这些结果表明，TRPV 4、TRPV 3和TRPM 8蛋白，而不是它们的离子通道活性，对于在32 ℃诱导CIP是必需的。鉴定在37 ℃和32 ℃下与这些TRP通道差异相互作用的蛋白质将有助于阐明低温诱导CIP的潜在机制。(C)2017作者由爱思唯尔公司出版
Cold-inducible RNA-binding protein (CIRP), RNA-binding motif protein 3 (RBM3) and serine and arginine rich splicing factor 5 (SRSF5) are RNA-binding proteins that are transcriptionally upregulated in response to moderately low temperatures and a variety of cellular stresses in mammalian cells. Induction of these cold-inducible proteins (CIPs) is dependent on transient receptor potential (TRP) V4 channel protein, but seems independent of its ion channel activity. We herein report that in addition to TRPV4, TRPV3 and TRPM8 are necessary for the induction of CIPs. We established cell lines from the lung of TRPV4-knockout (KO) mouse, and observed induction of CIPs in them by western blot analysis. A TRPV4 antagonist RN1734 suppressed the induction in wild-type mouse cells, but not in TRPV4-KO cells. A TRPV3 channel blocker 5408271 and a TRPM8 channel blocker AMTB as well as siRNAs against TRPV3 and TRPM8 suppressed the CIP induction in mouse TRPV4-KO cells and human U-2 OS cells. A TRPV3 channel agonist 2-APB induced CIP expression, but camphor did not. Neither did a TRPM8 channel agonist WS-12. These results suggest that TRPV4, TRPV3 and TRPM8 proteins, but not their ion channel activities are necessary for the induction of CIPs at 32 degrees C. Identification of proteins that differentially interact with these TRP channels at 37 degrees C and 32 degrees C would help elucidate the underlying mechanisms of CIP induction by hypothermia. (C) 2017 The Authors. Published by Elsevier Inc.