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DEC1 Modulates the Circadian Phase of Clock Gene Expression

机译：DEC1调节时钟基因表达的昼夜节律

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DEC1 suppresses CLOCK/BMAL1-enhanced promoter activity, but its role in the circadian system of mammals remains unclear. Here we examined the effect of Dec1 overexpression or deficiency on circadian gene expression triggered with 50% serum. Overexpression of Dec1 delayed the phase of clock genes such as Dec1, Dec2, Per1, and Dbp that contain E boxes in their regulatory regions, whereas it had little effect on the circadian phase of Per2 and Cry1 carrying CACGTT E′ boxes. In contrast, Dec1 deficiency advanced the phase of the E-box-containing clock genes but not that of the E′-box-containing clock genes. Accordingly, DEC1 showed strong binding and transrepression on the E box, but not on the E′ box, in chromatin immunoprecipitation, electrophoretic mobility shift, and luciferase reporter assays. Dec1?/? mice showed behavioral rhythms with slightly but significantly longer circadian periods under conditions of constant darkness and faster reentrainment to a 6-h phase-advanced shift of a light-dark cycle. Knockdown of Dec2 with small interfering RNA advanced the phase of Dec1 and Dbp expression, and double knockdown of Dec1 and Dec2 had much stronger effects on the expression of the E-box-containing clock genes. These findings suggest that DEC1, along with DEC2, plays a role in the finer regulation and robustness of the molecular clock.
机译：DEC1抑制CLOCK / BMAL1增强的启动子活性，但尚不清楚其在哺乳动物昼夜节律系统中的作用。在这里，我们研究了 Dec1 的过表达或缺乏对50％血清触发的昼夜节律基因表达的影响。 Dec1 的过表达延迟了时钟基因的相位，例如 Dec1 ， Dec2 ， Per1 和 Dbp 在其调控区域内包含E框，而对带有CACGTT E'框的 Per2 和 Cry1 的昼夜节律影响不大。相比之下， Dec1 缺陷使含E-box的时钟基因的相位提前，但不使含E'-box的时钟基因的相位提前。因此，在染色质免疫沉淀，电泳迁移率变化和萤光素酶报告基因检测中，DEC1在E盒上显示出很强的结合力，但在E'盒上却没有。 Dec1 ？/？小鼠表现出行为节律，在恒定的黑暗条件下，其昼夜节律周期略长，但明显更长，并且能够更快地将其重新夹带到6h相位提前的光-移相中。黑暗周期。用小干扰RNA敲除 Dec2 会促进 Dec1 和 Dbp 表达的阶段，而 Dec1 和 Dec2 对含有E-box的时钟基因的表达具有更强的影响。这些发现表明，DEC1和DEC2在分子时钟的更精细调节和鲁棒性中起作用。

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