Zhu Jingui, Sun Yongqiao, Zhao Fa-Qing, Yu Jun, Craig Roger, Hu Songnian
Key laboratory of Genome Sciences and Information, Beijing Institute of Genomics, Chinese Academy of Sciences, Beijing, PR China.
BMC Genomics. 2009 Mar 19;10:117. doi: 10.1186/1471-2164-10-117.
Tarantula has been used as a model system for studying skeletal muscle structure and function, yet data on the genes expressed in tarantula muscle are lacking.
We constructed a cDNA library from Aphonopelma sp. (Tarantula) skeletal muscle and got 2507 high-quality 5'ESTs (expressed sequence tags) from randomly picked clones. EST analysis showed 305 unigenes, among which 81 had more than 2 ESTs. Twenty abundant unigenes had matches to skeletal muscle-related genes including actin, myosin, tropomyosin, troponin-I, T and C, paramyosin, muscle LIM protein, muscle protein 20, a-actinin and tandem Ig/Fn motifs (found in giant sarcomere-related proteins). Matches to myosin light chain kinase and calponin were also identified. These results support the existence of both actin-linked and myosin-linked regulation in tarantula skeletal muscle. We have predicted full-length as well as partial cDNA sequences both experimentally and computationally for myosin heavy and light chains, actin, tropomyosin, and troponin-I, T and C, and have deduced the putative peptides. A preliminary analysis of the structural and functional properties was also carried out. Sequence similarities suggested multiple isoforms of most myofibrillar proteins, supporting the generality of multiple isoforms known from previous muscle sequence studies. This may be related to a mix of muscle fiber types.
The present study serves as a basis for defining the transcriptome of tarantula skeletal muscle, for future in vitro expression of tarantula proteins, and for interpreting structural and functional observations in this model species.
狼蛛已被用作研究骨骼肌结构和功能的模型系统,但关于狼蛛肌肉中表达基因的数据尚缺。
我们构建了来自墨西哥金背狼蛛(Aphonopelma sp.)骨骼肌的cDNA文库,并从随机挑选的克隆中获得了2507个高质量的5'端表达序列标签(EST)。EST分析显示有305个单基因,其中81个有超过2个EST。20个丰富的单基因与骨骼肌相关基因匹配,包括肌动蛋白、肌球蛋白、原肌球蛋白、肌钙蛋白I、T和C、副肌球蛋白、肌肉LIM蛋白、肌肉蛋白20、α - 辅肌动蛋白以及串联免疫球蛋白/纤连蛋白基序(在巨型肌节相关蛋白中发现)。还鉴定出了与肌球蛋白轻链激酶和钙调蛋白的匹配。这些结果支持狼蛛骨骼肌中存在肌动蛋白相关调节和肌球蛋白相关调节。我们通过实验和计算预测了肌球蛋白重链和轻链、肌动蛋白、原肌球蛋白以及肌钙蛋白I、T和C的全长及部分cDNA序列,并推导了假定的肽段。还对其结构和功能特性进行了初步分析。序列相似性表明大多数肌原纤维蛋白存在多种异构体,这支持了先前肌肉序列研究中已知的多种异构体的普遍性。这可能与肌肉纤维类型的混合有关。
本研究为定义狼蛛骨骼肌的转录组、未来狼蛛蛋白的体外表达以及解释该模型物种的结构和功能观察结果奠定了基础。