Bartoloni L, Blouin J L, Maiti A K, Sainsbury A, Rossier C, Gehrig C, She J X, Marron M P, Lander E S, Meeks M, Chung E, Armengot M, Jorissen M, Scott H S, Delozier-Blanchet C D, Gardiner R M, Antonarakis S E
Division of Medical Genetics, University of Geneva Medical School and, Geneva, Switzerland.
Genomics. 2001 Feb 15;72(1):21-33. doi: 10.1006/geno.2000.6462.
Dyneins are multisubunit protein complexes that couple ATPase activity with conformational changes. They are involved in the cytoplasmatic movement of organelles (cytoplasmic dyneins) and the bending of cilia and flagella (axonemal dyneins). Here we present the first complete cDNA and genomic sequences of a human axonemal dynein beta heavy chain gene, DNAH9, which maps to 17p12. The 14-kb-long cDNA is divided into 69 exons spread over 390 kb. The cDNA sequence of DNAH9 was determined using a combination of methods including 5' rapid amplification of cDNA ends, RT-PCR, and cDNA library screening. RT-PCR using nasal epithelium and testis RNA revealed several alternatively spliced transcripts. The genomic structure was determined using three overlapping BACs sequenced by the Whitehead Institute/MIT Center for Genome Research. The predicted protein, of 4486 amino acids, is highly homologous to sea urchin axonemal beta heavy chain dyneins (67% identity). It consists of an N-terminal stem and a globular C-terminus containing the four P-loops that constitute the motor domain. Lack of proper ciliary and flagellar movement characterizes primary ciliary dyskinesia (PCD), a genetically heterogeneous autosomal recessive disorder with respiratory tract infections, bronchiectasis, male subfertility, and, in 50% of cases, situs inversus (Kartagener syndrome, KS). Dyneins are excellent candidate genes for PCD and KS because in over 50% of cases the ultrastructural defects of cilia are related to the dynein complex. Genotype analysis was performed in 31 PCD families with two or more affected siblings using a highly informative dinucleotide polymorphism located in intron 26 of DNAH9. Two families with concordant inheritance of DNAH9 alleles in affected individuals were observed. A mutation search was performed in these two "candidate families," but only polymorphic variants were found. In the absence of pathogenic mutations, the DNAH9 gene has been excluded as being responsible for autosomal recessive PCD in these families.
动力蛋白是多亚基蛋白质复合物，它将ATP酶活性与构象变化联系起来。它们参与细胞器的细胞质运动（细胞质动力蛋白）以及纤毛和鞭毛的弯曲（轴丝动力蛋白）。在此，我们展示了人类轴丝动力蛋白β重链基因DNAH9的首个完整cDNA和基因组序列，该基因定位于17p12。这个14kb长的cDNA被分为69个外显子，分布在390kb的区域。DNAH9的cDNA序列是通过多种方法确定的，包括5' cDNA末端快速扩增、逆转录聚合酶链反应（RT-PCR）和cDNA文库筛选。使用鼻上皮和睾丸RNA进行的RT-PCR揭示了几种可变剪接转录本。基因组结构是通过对由怀特黑德研究所/麻省理工学院基因组研究中心测序的三个重叠细菌人工染色体（BAC）来确定的。预测的蛋白质有4486个氨基酸，与海胆轴丝β重链动力蛋白高度同源（同一性为67%）。它由一个N端茎和一个球状C端组成，C端包含构成运动结构域的四个P环。原发性纤毛运动障碍（PCD）的特征是缺乏正常的纤毛和鞭毛运动，这是一种遗传异质性常染色体隐性疾病，伴有呼吸道感染、支气管扩张、男性生育力低下，并且在50%的病例中存在内脏反位（卡塔格内综合征，KS）。动力蛋白是PCD和KS的优秀候选基因，因为在超过50%的病例中，纤毛的超微结构缺陷与动力蛋白复合物有关。使用位于DNAH9第26内含子中的一个信息丰富的二核苷酸多态性，对31个有两个或更多患病同胞的PCD家族进行了基因型分析。观察到两个家族中患病个体的DNAH9等位基因呈一致遗传。在这两个“候选家族”中进行了突变搜索，但只发现了多态性变体。在没有致病突变的情况下，DNAH9基因已被排除是这些家族中常染色体隐性PCD的病因。
