A rare variant in the osteoarthritis-associated locus GDF5 is functional and reveals a site that can be manipulated to modulate GDF5 expression.
A rare variant in the osteoarthritis-associated locus GDF5 is functional and reveals a site that can be manipulated to modulate GDF5 expression.
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DOI：
10.1038/ejhg.2012.197
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发表时间：
2013-05
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影响因子：
--
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骨关节炎 (OA) 是一种以软骨损失为特征的多基因疾病，单核苷酸多态性 (SNP) rs143383 (C/T) 影响多个种族群体的 OA 易感性。 SNP 位于生长和分化因子 5 基因 (GDF5) 的 5'-UTR 内，OA 相关 T 等位基因介导 GDF5 表达减少。由于 GDF5 编码软骨合成代谢蛋白，这种表达减少可以解释为什么 rs143383 的 T 等位基因是 OA 危险因素。我们对 GDF5 的深度测序发现了一个 C/A 颠换，位置相对于该基因的转录起始位点为-41bp。该启动子变体预计会影响转录因子结合，因此可能会突出显示一个可用于操纵 GDF5 表达并减轻 rs143383 的 T 等位基因介导的有害影响的调节位点。在这里，我们描述了对−41 bp变体的功能评估。使用报告基因构建体，我们证明颠换导致基因表达增加到A等位基因能够补偿由rs143383的T等位基因介导的表达减少的程度。使用电泳迁移率变动分析，我们确定 YY1 是一种反式作用因子，它与 -41bp 变体的等位基因有差异性结合，与等位基因 A 的结合更强烈。YY1 的敲低导致 GDF5 表达显着减少，支持 YY1 作为 GDF5 激活剂。总之，我们证明了-41bp变体是有功能的，并且我们已经确定了GDF5的一个调节区域，可以用来克服由rs143383的T等位基因介导的OA遗传缺陷。
Osteoarthritis (OA) is a polygenic disease characterized by cartilage loss, with the single-nucleotide polymorphism (SNP) rs143383 (C/T) influencing OA susceptibility across a range of ethnic groups. The SNP resides within the 5′-UTR of the growth and differentiation factor 5 gene (GDF5), with the OA-associated T-allele mediating reduced GDF5 expression. As GDF5 codes for a cartilage anabolic protein, this reduced expression may explain why the T-allele of rs143383 is an OA risk factor. Our deep sequencing of GDF5 identified a C/A transversion located −41 bp relative to the gene's transcription start site. This promoter variant is predicted to affect transcription factor binding and it may therefore highlight a regulatory site that could be exploited to manipulate GDF5 expression and alleviate the detrimental effect mediated by the T-allele of rs143383. Here, we describe our functional assessment of the −41 bp variant. Using reporter constructs we demonstrated that the transversion leads to increased gene expression to such a degree that the A-allele is able to compensate for the reduced expression mediated by the T-allele of rs143383. Using electrophoretic mobility shift assays we identified YY1 as a trans-acting factor that differentially binds to the alleles of the −41 bp variant, with more avid binding to allele A. Knockdown of YY1 led to a significant reduction in GDF5 expression, supporting YY1 as a GDF5 activator. In conclusion, we demonstrated that the −41 bp variant is functional and we have identified a regulatory region of GDF5 that can be exploited to overcome the OA genetic deficit mediated by the T-allele of rs143383.
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DOI：
10.1126/science.1219240
发表时间：
2012-07-06
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Tennessen JA;Bigham AW;O'Connor TD;Fu W;Kenny EE;Gravel S;McGee S;Do R;Liu X;Jun G;Kang HM;Jordan D;Leal SM;Gabriel S;Rieder MJ;Abecasis G;Altshuler D;Nickerson DA;Boerwinkle E;Sunyaev S;Bustamante CD;Bamshad MJ;Akey JM;Broad GO;Seattle GO;NHLBI Exome Sequencing Project
通讯作者：
NHLBI Exome Sequencing Project
影响因子：
30.8
作者：
Thomas, JT;Kilpatrick, MW;Luyten, FP
通讯作者：
Luyten, FP
影响因子：
30.8
作者：
通讯作者：
--
影响因子：
3.5
作者：
Chapman, Kay;Takahashi, Atsushi;Ikegawa, Shiro
通讯作者：
Ikegawa, Shiro
影响因子：
--
作者：
Egli RJ;Southam L;Wilkins JM;Lorenzen I;Pombo-Suarez M;Gonzalez A;Carr A;Chapman K;Loughlin J
通讯作者：
Loughlin J