Characteristics of a Critical Calcium Transportation Regulator: Plasma Membrane Ca2+-ATPase Involved in Calcium Homeostasis from Hyriopsis cumingii (Lea)

A. Zhang, Zhiming Zhou
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Abstract

Plasma Membrane Calcium ATPase (PMCA) plays a critical role in transporting Ca2+ out of the cytosol across the plasma membrane. Here, a full-length cDNA sequence of plasma membrane Ca2+-ATPase gene was isolated from the gill of Hyriopsis cumingii (HcPMCA) by using SMART RACE technique. The entire cDNA was 5230 bp, including a 417-bp 5'-UTR, a 3588-bp ORF and a 1225-bp 3'-UTR, encoding a 1195-amino acid protein, and no putative signal peptide was predicted. Compared with PMCA homologs from seawater mollusks, HcPMCA had high similarity with them in both sequence and structure. Tissue-specific expression analysis revealed that HcPMCA mRNA was detected in all the sampled tissues, but was prominently expressed in the gill and mantle. When exposed to a serie of increasing Ca2+ that lasted for 7 days, the mRNA expression of HcPMCA in the mantle was slightly downregulated, but peaked at 60 mg/L. Moreover, the temporal expression of HcPMCA transcripts in the mantle after 60 mg/L Ca2+ exposure was shown to be bell-shaped, which was slightly downregulated at 24 h, but upregulated from 24 h to 48 h post-treatment, peaking at 48 h. The result of present study provides useful information for further studies on function and regulation mechanism of HcPMCA gene.
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一种关键钙转运调节因子:三角帆蚌(Lea)质膜Ca2+-ATP酶参与钙稳态的特性
质膜钙ATP酶(PMCA)在将Ca2+转运出胞质溶胶穿过质膜方面起着关键作用。本文利用SMART RACE技术从三角帆蚌(HcPMCA)鳃中分离到一个质膜Ca2+-ATP酶基因的全长cDNA序列。整个cDNA全长5230bp,包括417bp的5'-UTR、3588bp的ORF和1225bp的3'-UTR,编码1195个氨基酸的蛋白质,并且没有预测到推定的信号肽。与海水软体动物的PMCA同源物相比,HcPMCA在序列和结构上与它们具有高度相似性。组织特异性表达分析显示,HcPMCA mRNA在所有取样组织中都检测到,但在鳃和地幔中显著表达。当暴露于持续7天的一系列增加的Ca2+时,地幔中HcPMCA的mRNA表达略有下调,但在60mg/L时达到峰值。此外,60 mg/L Ca2+暴露后,HcPMCA转录物在地幔中的时间表达呈钟形,在24小时略有下调,但在处理后24小时至48小时上调,在48小时达到峰值。本研究的结果为进一步研究HcPMCA基因的功能和调节机制提供了有用的信息。
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