青光眼相关的WDR36变体在酵母模型系统中编码功能缺陷。
Glaucoma-associated WDR36 variants encode functional defects in a yeast model system.
作者信息
Footz Tim K, Johnson Jill L, Dubois Stéphane, Boivin Nicolas, Raymond Vincent, Walter Michael A
机构信息
Department of Medical Genetics, University of Alberta, Edmonton, AB, Canada.
出版信息
Hum Mol Genet. 2009 Apr 1;18(7):1276-87. doi: 10.1093/hmg/ddp027. Epub 2009 Jan 15.
Primary open-angle glaucoma (POAG) is a leading cause of blindness worldwide. POAG is associated with a characteristic progression of changes to ocular morphology and degeneration at the optic nerve head with the loss of visual fields. Physical mapping efforts identified genomic loci in which to search for causative POAG gene mutations. WDR36, at locus GLC1G, was initially identified as a gene with a low frequency of non-synonymous sequence variations that were exclusive to adult-onset POAG patients. It has since been shown that rare WDR36 sequence variants are also present in the normal population at similarly low frequencies. The lack of a consistent genotype:phenotype correlation prompted us to investigate the functional consequences of WDR36 sequence variations. WDR36 is involved in rRNA processing, a critical step in ribosome biogenesis, and is very similar to yeast Utp21p which is a member of the small subunit (SSU) processome complex responsible for maturation of 18S rRNA. We, therefore, developed a yeast model system to test the functional and phenotypic consequences of POAG-associated sequence variants introduced into UTP21. Alone, the POAG variants did not produce any significant defects in cell viability or rRNA processing. However, when combined with disruption of STI1 (which synthetically interacts with UTP21), 5 of the 11 tested variants had increased or decreased cell viability which corresponded to reduced or elevated levels of pre-rRNA, respectively. These results demonstrate that, in the correct genetic background, WDR36 sequence variants can lead to an altered cellular phenotype, supporting the theory that WDR36 participates in polygenic forms of glaucoma.
原发性开角型青光眼(POAG)是全球失明的主要原因。POAG与眼部形态特征性变化的进展以及视神经乳头的退化和视野丧失有关。物理图谱研究确定了基因组位点,以便在其中寻找导致POAG的基因突变。位于GLC1G位点的WDR36最初被鉴定为一个基因,其非同义序列变异频率较低,且仅见于成年发病的POAG患者。此后发现,正常人群中也存在频率同样较低的罕见WDR36序列变异。由于缺乏一致的基因型与表型相关性,我们对WDR36序列变异的功能后果进行了研究。WDR36参与rRNA加工,这是核糖体生物合成中的关键步骤,并且与酵母Utp21p非常相似,后者是负责18S rRNA成熟的小亚基(SSU)加工体复合物的成员。因此,我们开发了一个酵母模型系统,以测试引入UTP21的POAG相关序列变异的功能和表型后果。单独来看,POAG变异在细胞活力或rRNA加工方面未产生任何显著缺陷。然而,当与STI1(与UTP21存在合成相互作用)的破坏相结合时,11个测试变异中的5个导致细胞活力增加或降低,分别对应于前体rRNA水平的降低或升高。这些结果表明,在正确的遗传背景下,WDR36序列变异可导致细胞表型改变,支持WDR36参与青光眼多基因形式的理论。