Bhatia-Dey N, Adelstein R S, Dawid I B
Laboratory of Molecular Genetics, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD 20892.
Proc Natl Acad Sci U S A. 1993 Apr 1;90(7):2856-9. doi: 10.1073/pnas.90.7.2856.
The complete amino acid sequence of Xenopus laevis nonmuscle myosin heavy chain B (MHC-B) has been deduced from overlapping cDNA clones isolated from an XTC cell library. RNA blots of various developmental stages, adult tissues, and XTC cells detect a single transcript of 7.5 kb which is expressed at similar levels throughout development. MHC-B mRNA was detected in XTC cells, heart, lung, spleen, and brain, at lower levels in ovary, testis, pancreas, stomach, liver, and eye, but not in kidney and skeletal muscle. Protein expression in adult tissues, as detected by immunoblot analysis, correlates well with mRNA expression. In chickens and humans, a fraction of the mRNA encoding the MHC-B isoform was found previously to contain a 10-amino acid insert at amino acid 211 near the ATP-binding site. As reported elsewhere, in the chicken this insert-bearing isoform is nervous system-specific. The Xenopus sequence shows a 16-amino acid insertion at the same position; 7 of 16 residues are identical to those in the chicken and human insertion, and these identical residues include a consensus target sequence for cyclin-p34cdc2 kinase. In contrast to chicken, all frog tissues and embryonic stages tested contained the insert-bearing form, and no evidence for a non-insert-bearing MHC-B isoform was found in Xenopus.
从非洲爪蟾XTC细胞文库中分离出的重叠cDNA克隆已推导得到非洲爪蟾非肌肉肌球蛋白重链B(MHC - B)的完整氨基酸序列。对不同发育阶段、成体组织和XTC细胞进行RNA印迹分析,检测到一条7.5 kb的单一转录本,其在整个发育过程中表达水平相似。在XTC细胞、心脏、肺、脾脏和大脑中检测到MHC - B mRNA,在卵巢、睾丸、胰腺、胃、肝脏和眼睛中表达水平较低,但在肾脏和骨骼肌中未检测到。通过免疫印迹分析检测到的成体组织中的蛋白质表达与mRNA表达良好相关。在鸡和人类中,先前发现编码MHC - B同工型的一部分mRNA在靠近ATP结合位点的氨基酸211处含有一个10个氨基酸的插入序列。如其他地方所报道的,在鸡中,这种带有插入序列的同工型是神经系统特异性的。非洲爪蟾的序列在相同位置显示有一个16个氨基酸的插入;16个残基中有7个与鸡和人类插入序列中的残基相同,并且这些相同的残基包括细胞周期蛋白 - p34cdc2激酶的共有靶序列。与鸡不同,所检测的所有蛙类组织和胚胎阶段都含有带有插入序列的形式,并且在非洲爪蟾中未发现无插入序列的MHC - B同工型的证据。