Miyatani S, Winkles J A, Sargent T D, Dawid I B
J Cell Biol. 1986 Nov;103(5):1957-65. doi: 10.1083/jcb.103.5.1957.
This report describes the isolation and characterization of genomic and cDNA clones which define a subfamily of type I keratins in Xenopus laevis whose expression is restricted to embryonic and larval stages. The XK81 subfamily, named after the prototype cDNA clone DG81, contains four members arranged in two pairs of closely homologous loci; they were named 81A1, A2, B1, and B2. Genomic clones were obtained representing all of these regions. The A1 gene has been completely sequenced together with approximately 1 kb of flanking sequences at each end; this gene corresponds to the previously reported cDNA clone 8128 (Jonas, E., T. D. Sargent, and I. B. Dawid, 1985, Proc. Natl. Acad. Sci. USA, 82:5413-5417). The B2 gene is represented by a partial cDNA clone, DG118. Upstream sequences and about half of the coding regions have been sequenced for the B1 and B2 genes, whereas the A2 locus has been identified on the basis of hybridization data and could be a gene or pseudogene. Genomic Southern blotting indicates that all members of the subfamily have been isolated. The keratin proteins encoded by the B1 and B2 genes are 96% homologous in the central rod domain, whereas A/B gene homology in this region is 81%. During development mRNAs derived from A and B genes accumulate coordinately during gastrula and neurula stages; in the tadpole, 81A mRNA decays rapidly, whereas 81B mRNA shows a second abundance peak, persists for most of tadpole life, and decays by metamorphosis. RNAs derived from the XK81 keratin subfamily are undetectable in the adult, where different type I keratin genes are expressed.
本报告描述了基因组和cDNA克隆的分离与鉴定,这些克隆定义了非洲爪蟾I型角蛋白的一个亚家族,其表达仅限于胚胎和幼虫阶段。以原型cDNA克隆DG81命名的XK81亚家族包含四个成员,排列成两对紧密同源的基因座;它们被命名为81A1、A2、B1和B2。获得了代表所有这些区域的基因组克隆。A1基因已完全测序,两端各有约1 kb的侧翼序列;该基因对应于先前报道的cDNA克隆8128(乔纳斯,E.,T. D. 萨金特,和I. B. 戴维德,1985,《美国国家科学院院刊》,82:5413 - 5417)。B2基因由一个部分cDNA克隆DG118代表。B1和B2基因的上游序列和大约一半的编码区已测序,而A2基因座是根据杂交数据鉴定的,可能是一个基因或假基因。基因组Southern印迹分析表明该亚家族的所有成员均已分离。B1和B2基因编码的角蛋白在中央杆状结构域的同源性为96%,而该区域A/B基因的同源性为81%。在发育过程中,A和B基因的mRNA在原肠胚和神经胚阶段协同积累;在蝌蚪中,81A mRNA迅速降解,而81B mRNA出现第二个丰度峰值,在蝌蚪的大部分生命期持续存在,并在变态时降解。在成体中检测不到来自XK81角蛋白亚家族的RNA,成体中表达的是不同的I型角蛋白基因。