Polymorphism in the PER3 Promoter Associates with Diurnal Preference and Delayed Sleep Phase Disorder
Polymorphism in the PER3 Promoter Associates with Diurnal Preference and Delayed Sleep Phase Disorder
复制标题
DOI：
10.1093/sleep/33.5.695
复制
发表时间：
2010-05-01
期刊：
影响因子：
5.6
通讯作者：
von Schantz, Malcolm
中科院分区：
文献类型：
--
作者：
Archer, Simon N.;Carpen, Jayshan D.;von Schantz, Malcolm
研究目的：筛选PER 3启动子的多态性，并调查这些多态性与昼夜偏好，睡眠期延迟障碍/综合征（DSPD/DSPS），和它们对报告基因expression.Design的影响的表型协会：种间比较被用来定义的PER 3启动子的转录起始位点和核苷酸位置-874之间的区域的近似程度。使用PCR和直接测序在DNA池中筛选该区域，这也用于筛选来自个体参与者的DNA。将不同的启动子等位基因克隆到荧光素酶表达载体中并创建缺失文库。启动子激活用化学发光法测定。设置：N/AP患者或参与者：DNA样本来自具有确定的昼夜偏好的志愿者（3 x 80，选自1，590例患者）和DSPD患者（n = 23）。干预：N/A测量和结果：我们验证了3个单核苷酸多态性（G-320 T，C-319 A，G-294 A），并发现了一个新的可变数目串联重复序列（VNTR）多态性（-318 1/2 VNTR）。DSPD中-320T和-319A等位基因的频率高于早晨型（P = 0.042）和晚上型（P = 0.006和0.033）。等位基因组合TA 2G在DSPD中比早晨型（P = 0.033）或晚上型（P = 0.002）更普遍。由TA 2G组合驱动的荧光素酶表达大于更常见的GC 2A（P < 0.05）和更罕见的TA 1G（P < 0.001）组合。缺失报告构建体鉴定了两个增强子区域（-703至-605和-283至-80）。结论：PER 3启动子的多态性可能影响其表达，导致观察到的PER 3功能的潜在差异。
Study Objectives: To screen the PER3 promoter for polymorphisms and investigate the phenotypic associations of these polymorphisms with diurnal preference, delayed sleep phase disorder/syndrome (DSPD/DSPS), and their effects on reporter gene expression.Design: Interspecific comparison was used to define the approximate extent of the PER3 promoter as the region between the transcriptional start site and nucleotide position -874. This region was screened in DNA pools using PCR and direct sequencing, which was also used to screen DNA from individual participants. The different promoter alleles were cloned into a luciferase expression vector and a deletion library created. Promoter activation was measured by chemiluminescence.Setting: N/APatients or Participants: DNA samples were obtained from volunteers with defined diurnal preference (3 x 80, selected from a pool of 1,590), and DSPD patients (n = 23).Interventions: N/AMeasurements and Results: We verified three single nucleotide polymorphisms (G -320T, C -319A, G -294A), and found a novel variable number tandem repeat (VNTR) polymorphism (-318 1/2 VNTR). The -320T and -319A alleles occurred more frequently in DSPD compared to morning (P = 0.042 for each) or evening types (P = 0.006 and 0.033). The allele combination TA2G was more prevalent in DSPD compared to morning (P = 0.033) or evening types (P = 0.002). Luciferase expression driven by the TA2G combination was greater than for the more common GC2A (P < 0.05) and the rarer TA1G (P < 0.001) combinations. Deletion reporter constructs identified two enhancer regions (-703 to -605, and -283 to -80).Conclusions: Polymorphisms in the PER3 promoter could affect its expression, leading to potential differences in the observed functions of PER3.