Isolation of several human axonemal dynein heavy chain genes: genomic structure of the catalytic site, phylogenetic analysis and chromosomal assignment

Abstract

Dynein heavy chains (DHCs) are the main components of multisubunit motor ATPase complexes called dyneins. Axonemal dyneins provide the driving force for ciliary and flagellar motility. Recent molecular studies demonstrated that multiple DHC isoforms are produced by separate genes. We describe the isolation of five human axonemal DHC genes. Analysis of the human genomic clones revealed the existence of intronic sequences that were used to demonstrate that human axonemal DHC genes are located on different chromosomes. The cloned human DHC sequences were integrated into an evolutionary approach based on phylogenetic analysis. Tissue expression studies showed that these human axonemal DHCs are expressed in testis and/or trachea, two tissues with axonemal structures that can be altered in primary ciliary dyskinesia, making DHC genes strong candidates in the genesis of these human diseases.

Identification, tissue specific expression, and chromosomal localisation of several human dynein heavy chain genes

“…Specific upstream primers were designed, pooled and used with DNAHpool-4R in an attempt to amplify dynein fragments for PAC clones. Upstream specific primers are: RT-DNAH1F; DNAH-2F, 14 RT-DNAH3F, RT-DNAH5F, DNAH-6Fbis, RT-DNAH7F, RT-DNAH9F, RT-DNAH10F, RT-DNAH11F, RT-DNAH12F, 16 RT-DNAH14F. 14 Three PAC clones amplified a 276 bp fragment that after cloning and sequencing of several clones corresponded to (HSA)DNAH3.…”

“…6 Numerous distinct dynein heavy chain genes have been isolated from Paramecium, 7 Chlamydomonas, 8,9 sea urchin, 10 rat, 11,12 mouse, 13 and human. [13][14][15][16] However, comparing the dynein heavy chain gene family in well characterised species such as sea urchin suggests that either mammals have less axonemal dynein heavy genes or that some mammal genes are still to be discovered.…”

Alterations of the spindle checkpoint pathway in clinicopathologically aggressive CpG island methylator phenotype clear cell renal cell carcinomas

“…Seventy‐six genes showed a genetic aberration incidence of 10% or more in CIMP‐positive RCCs ( n  = 19), whereas only four genes did so in CIMP‐negative RCCs ( n  = 87) (Tables 2 and 3). Genes encoding microtubule‐associated proteins, such as DNAH2, DNAH5, DNAH10 ,31 RP1 32 and HAUS8 ,33, 34 those involved in histone modification, such as NCOA1 ,35 those involved in cell adhesion, such as CELSR1, CELSR2 ,36 CTNND1 ,37 LAMC2 38 and TJP1 ,39 and tumor‐related genes such as BAP1 40 and ATM ,41 were frequently mutated in CIMP‐positive RCCs (Table 3). As shown in Tables 2 and 3, 235 genetic aberrations (173 non‐synonymous single‐nucleotide mutations and 62 indels) revealed by whole‐exome analysis in the initial cohort were all successfully verified by Sanger sequencing.…”

“…Aberrations of genes involved in cell adhesion, such as CELSR1, CELSR2 ,36 CTNND1 ,37 LAMC2 38 and TJP1 ,39 may affect the invasiveness and metastatic potential of CIMP‐positive RCCs (CIMP‐positive RCCs show invasive growth and distant metastasis more frequently than CIMP‐negative RCCs13). Moreover, genetic aberrations of microtubule‐associated proteins, such as DNAH2, DNAH5, DNAH10 ,31 RP1 32 and HAUS8 ,33, 34 may be correlated with dysregulation of the spindle checkpoint in CIMP‐positive RCCs. DNAH2, DNAH5 and DNAH10 encode the heavy chains of axonal dynein 31.…”

“…Moreover, genetic aberrations of microtubule‐associated proteins, such as DNAH2, DNAH5, DNAH10 ,31 RP1 32 and HAUS8 ,33, 34 may be correlated with dysregulation of the spindle checkpoint in CIMP‐positive RCCs. DNAH2, DNAH5 and DNAH10 encode the heavy chains of axonal dynein 31. In mice, mutations of the axonal dynein gene Left‐right dynein (LRD) result in abnormal segregation of sister chromatids,49 suggesting that axonal dynein may be involved in the movement of chromosomes and positioning of the mitotic spindles for cell division.…”