Garber A T, Retief J D, Dixon G H
Department of Medical Biochemistry, University of Calgary, Alberta, Canada.
EMBO J. 1989 Jun;8(6):1727-34. doi: 10.1002/j.1460-2075.1989.tb03565.x.
A Mg2+-dependent ATPase activity has been purified from trout sperm axonemes which has properties characteristic of a dynein ATPase. A polyclonal antiserum prepared against the dynein heavy chains has been used to isolate dynein heavy chain (DYHC) cDNAs from a trout testis lambda gt11 cDNA expression library. beta-galactosidase fusion proteins produced in lambda gt11 by these trout cDNAs cross-reacted with a heterologous anti-sea urchin dynein antiserum. Northern blot analyses demonstrated that the RNA transcripts detected have sizes (7.5 - 12 kb) consistent with those expected for the dynein heavy chains. All the DYHC cDNAs encode portions of a highly unusual DNA coding sequence comprised of 21 bp direct repeats. The predicted open reading frame of this repeat is Ile/Leu-His-Val-Ile-Gln-Tyr-Ser and is characteristic of an extensive alpha-helical coiled-coil domain. The presence of an in-frame translation termination codon indicates that this domain is located at the carboxyl-terminus of the DYHC. Southern blot analyses demonstrated a low, if not single, copy number for this gene and conservation of this domain in other vertebrates. DYHC transcripts reach their highest level in testis, but are also abundant in brain tissue.
已从虹鳟鱼精子轴丝中纯化出一种依赖镁离子的ATP酶活性，其具有动力蛋白ATP酶的特性。针对动力蛋白重链制备的多克隆抗血清已用于从虹鳟鱼睾丸λgt11 cDNA表达文库中分离动力蛋白重链（DYHC）cDNA。这些虹鳟鱼cDNA在λgt11中产生的β-半乳糖苷酶融合蛋白与异源抗海胆动力蛋白抗血清发生交叉反应。Northern印迹分析表明，检测到的RNA转录本大小（7.5 - 12 kb）与动力蛋白重链预期大小一致。所有DYHC cDNA编码由21 bp直接重复序列组成的高度不寻常的DNA编码序列的部分。该重复序列的预测开放阅读框为Ile/Leu-His-Val-Ile-Gln-Tyr-Ser，是广泛的α-螺旋卷曲螺旋结构域的特征。框内翻译终止密码子的存在表明该结构域位于DYHC的羧基末端。Southern印迹分析表明该基因的拷贝数较低，甚至可能是单拷贝，且该结构域在其他脊椎动物中保守。DYHC转录本在睾丸中达到最高水平，但在脑组织中也很丰富。
