Thermally induced and developmentally regulated expression of a small heat shock protein in Trichinella spiralis
Thermally induced and developmentally regulated expression of a small heat shock protein in Trichinella spiralis
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DOI：
10.1007/s00436-007-0462-6
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发表时间：
2007-02
影响因子：
2
通讯作者：
Z. Wu;I. Nagano;T. Boonmars;Y. Takahashi
中科院分区：
文献类型：
--
作者：
Z. Wu;I. Nagano;T. Boonmars;Y. Takahashi
本文克隆并表达了旋毛虫热休克蛋白（Ts-sHsp）的cDNA。该cDNA编码一个165个氨基酸的预测蛋白，在α晶体蛋白结构域与其他生物的各种小分子热休克蛋白具有高度的序列同源性。Westernblot分析表明，抗Ts-sHsp重组抗体识别成虫和幼虫迁移的蛋白质，分子量约为19 kDa。原位定位研究表明，该蛋白大量存在于肌幼虫的体壁肌细胞、皮下组织、肌细胞和食道中。Ts-sHsp重组蛋白具有分子伴侣活性，可抑制柠檬酸合酶的热诱导聚集。该sHsp在T.螺旋，但在不同的水平。在成熟肌幼虫（感染性幼虫）中观察到高水平，远高于在成虫、新生幼虫或未成熟肌幼虫中观察到的水平。sHsp基因的表达是热诱导的，因此对冷（0°C）和热休克（43°C）胁迫都有响应;然而，以不同的模式。低温胁迫后3 ~ 72 h，Ts-sHsp的表达量逐渐增加，高温胁迫后3 h，Ts-sHsp的表达量最高，随后下降。这些结果表明，这种小分子热休克蛋白可能在旋毛虫幼虫对化学和物理胁迫的耐受中起作用，从而提高旋毛虫幼虫的存活能力。
A cDNA encoding a small heat shock protein ofTrichinella spiralis, Ts-sHsp, was cloned and expressed and is herein characterized. This cDNA encoded a predicted protein of 165 amino acids, which had a high sequence identity in α crystallin domain with various small heat shock proteins of other organisms. A Western blot analysis indicated that anti-Ts-sHsp recombinant antibody recognized the protein of adults and larvae migrating at about 19 kDa. An in situ localization study showed the protein to be abundantly present in the body wall muscle cells, hypodermis, stichocytes, and esophagus of muscle larvae. The Ts-sHsp recombinant protein possessed chaperone activity to suppress the thermally-induced aggregation of citrate synthase. This sHsp was expressed at various developmental stages ofT. spiralis, but at different levels. A high level was observed in mature muscle larvae (infective larvae), which was much higher than the levels seen in adults, newborn larvae, or immature muscle larvae. The expression of the sHsp gene was thermal inducible, thus responding to both cold (0°C) and heat shock (43°C) stress; however, at different patterns. The expression of Ts-sHsp increased gradually from 3 to 72 h after cold stress, while the expression was elevated to its highest after 3 h heat stress and then decreased. These results suggest that this small heat shock protein likely plays a role in the tolerance to both chemical and physical stresses, thereby enhancing the survival ability ofTrichinellamuscle larvae.