Kikuchi K, Muramatsu M, Hirayama Y, Watabe S
Laboratory of Aquatic Molecular Biology and Biotechnology, Graduate School of Agricultural and Life Sciences, The University of Tokyo, Bunkyo, Tokyo 113-8657, Japan.
Gene. 1999 Mar 4;228(1-2):189-96. doi: 10.1016/s0378-1119(99)00005-0.
We isolated partial coding sequences for 29 carp myosin heavy chain genes (MyoHCs) and determined the nucleotide sequences around the region encoding the loop 2 of the myosin molecule. The predicted amino acid sequences from the isolated genes all showed very high similarity to those of skeletal and cardiac muscles from higher vertebrates, but not to those of smooth and non-muscle counterparts. Among all clones isolated, carp MyoHC10, MyoHCI-1-3 and MyoHC30 showed exon-nucleotide sequences identical to those of cDNAs encoding the loop 2 region of the 10 degrees C-, intermediate- and 30 degrees C-type fast skeletal isoforms [Hirayama and Watabe, Euro. J. Biochem. 246 (1997) 380-387]. The loop 2 of 28 types of carp MyoHCs was encoded by two exons separated by an intron corresponding to that of the 16th in higher vertebrate MyoHCs, whilst this intron was not found in carp MyoHC30. Although carp MyoHC30 had a gene organization different from those of higher vertebrates and other carp MyoHCs, its predicted amino acid sequence for loop 2 showed the highest homology to those of higher vertebrates among carp MyoHCs. In the 28 carp MyoHCs containing the intron, a combination of different nucleotide sequences for the two resulted in 14 distinct series for the combined coding sequence. These different nucleotide sequences encoded nine distinct amino acid sequences. Phylogenetic analysis for the present loop 2 and light meromyosin previously reported for carp MyoHCs [Imai et al., J. Exp. Biol. 200 (1997) 27-34] revealed that carp MyoHCs have recently diverged and are more closely related to each other than to MyoHCs from other species.
我们分离出了29个鲤鱼肌球蛋白重链基因(MyoHCs)的部分编码序列,并确定了肌球蛋白分子环2编码区域周围的核苷酸序列。从分离出的基因预测的氨基酸序列与高等脊椎动物的骨骼肌和心肌的氨基酸序列都显示出非常高的相似性,但与平滑肌和非肌肉对应物的氨基酸序列不同。在所有分离出的克隆中,鲤鱼MyoHC10、MyoHCI-1-3和MyoHC30的外显子核苷酸序列与编码10℃型、中间型和30℃型快速骨骼肌同工型环2区域的cDNA的序列相同[平山和渡边,《欧洲生物化学杂志》246(1997)380 - 387]。28种鲤鱼MyoHCs的环2由两个外显子编码,这两个外显子被一个内含子隔开,该内含子与高等脊椎动物MyoHCs中第16个内含子相对应,而在鲤鱼MyoHC30中未发现这个内含子。尽管鲤鱼MyoHC30的基因结构与高等脊椎动物和其他鲤鱼MyoHCs不同,但其环2的预测氨基酸序列在鲤鱼MyoHCs中与高等脊椎动物的序列具有最高的同源性。在含有该内含子的28个鲤鱼MyoHCs中,两者不同的核苷酸序列组合产生了14个不同的组合编码序列系列。这些不同的核苷酸序列编码了9个不同的氨基酸序列。对目前的环2和先前报道的鲤鱼MyoHCs的轻酶解肌球蛋白进行系统发育分析[今井等人,《实验生物学杂志》200(1997)27 - 34]表明,鲤鱼MyoHCs最近发生了分化,它们彼此之间的关系比与其他物种的MyoHCs更为密切。