Tian Jianan, Keller Mark P, Oler Angie T, Rabaglia Mary E, Schueler Kathryn L, Stapleton Donald S, Broman Aimee Teo, Zhao Wen, Kendziorski Christina, Yandell Brian S, Hagenbuch Bruno, Broman Karl W, Attie Alan D
Department of Statistics, University of Wisconsin, Madison, Wisconsin 53706.
Department of Biochemistry, University of Wisconsin, Madison, Wisconsin 53706.
Genetics. 2015 Nov;201(3):1253-62. doi: 10.1534/genetics.115.179432. Epub 2015 Sep 18.
We surveyed gene expression in six tissues in an F2 intercross between mouse strains C57BL/6J (abbreviated B6) and BTBR T(+) tf/J (abbreviated BTBR) made genetically obese with the Leptin(ob) mutation. We identified a number of expression quantitative trait loci (eQTL) affecting the expression of numerous genes distal to the locus, called trans-eQTL hotspots. Some of these trans-eQTL hotspots showed effects in multiple tissues, whereas some were specific to a single tissue. An unusually large number of transcripts (∼8% of genes) mapped in trans to a hotspot on chromosome 6, specifically in pancreatic islets. By considering the first two principal components of the expression of genes mapping to this region, we were able to convert the multivariate phenotype into a simple Mendelian trait. Fine mapping the locus by traditional methods reduced the QTL interval to a 298-kb region containing only three genes, including Slco1a6, one member of a large family of organic anion transporters. Direct genomic sequencing of all Slco1a6 exons identified a nonsynonymous coding SNP that converts a highly conserved proline residue at amino acid position 564 to serine. Molecular modeling suggests that Pro564 faces an aqueous pore within this 12-transmembrane domain-spanning protein. When transiently overexpressed in HEK293 cells, BTBR organic anion transporting polypeptide (OATP)1A6-mediated cellular uptake of the bile acid taurocholic acid (TCA) was enhanced compared to B6 OATP1A6. Our results suggest that genetic variation in Slco1a6 leads to altered transport of TCA (and potentially other bile acids) by pancreatic islets, resulting in broad gene regulation.
我们对携带瘦素(ob)突变而导致遗传性肥胖的小鼠品系C57BL/6J(简称B6)和BTBR T(+) tf/J(简称BTBR)之间的F2杂交后代的六种组织中的基因表达进行了研究。我们鉴定出了一些影响基因座远端众多基因表达的表达数量性状基因座(eQTL),即反式eQTL热点。其中一些反式eQTL热点在多种组织中显示出效应,而有些则仅在单一组织中具有特异性。异常大量的转录本(约占基因的8%)反式定位到6号染色体上的一个热点,特别是在胰岛中。通过考虑定位到该区域的基因表达的前两个主成分,我们能够将多变量表型转化为简单的孟德尔性状。用传统方法对该基因座进行精细定位,将QTL区间缩小到一个仅包含三个基因的298 kb区域,其中包括有机阴离子转运蛋白大家族的一个成员Slco1a6。对所有Slco1a6外显子进行直接基因组测序,鉴定出一个非同义编码单核苷酸多态性,该多态性将氨基酸位置564处高度保守的脯氨酸残基转化为丝氨酸。分子模型表明,Pro564位于这个跨12个跨膜结构域的蛋白质内的一个水相孔中。当在HEK293细胞中瞬时过表达时,与B6 OATP1A6相比,BTBR有机阴离子转运多肽(OATP)1A6介导的胆汁酸牛磺胆酸(TCA)的细胞摄取增强。我们的结果表明,Slco1a6中的遗传变异导致胰岛对TCA(以及可能的其他胆汁酸)的转运改变,从而导致广泛的基因调控。