Shalev Moran, Aviram Rona, Adamovich Yaarit, Kraut-Cohen Judith, Shamia Tal, Ben-Dor Shifra, Golik Marina, Asher Gad
Department of Biological Chemistry, Weizmann Institute of Science, Rehovot 76100, Israel.
Biological Services, Weizmann Institute of Science, Rehovot 76100, Israel.
Nucleic Acids Res. 2014 Oct 29;42(19):11879-90. doi: 10.1093/nar/gku873. Epub 2014 Sep 26.
The circadian core clock circuitry relies on interlocked transcription-translation feedback loops that largely count on multiple protein interactions. The molecular mechanisms implicated in the assembly of these protein complexes are relatively unknown. Our bioinformatics analysis of short linear motifs, implicated in protein interactions, reveals an enrichment of the Pro-X-Asp-Leu-Ser (PXDLS) motif within circadian transcripts. We show that the PXDLS motif can bind to BMAL1/CLOCK and disrupt circadian oscillations in a cell-autonomous manner. Remarkably, the motif is evolutionary conserved in the core clock protein REV-ERBα, and additional proteins implicated in the clock's function (NRIP1, CBP). In this conjuncture, we uncover a novel cross talk between the two principal core clock feedback loops and show that BMAL/CLOCK and REV-ERBα interact and that the PXDLS motif of REV-ERBα participates in their binding. Furthermore, we demonstrate that the PXDLS motifs of NRIP1 and CBP are involved in circadian rhythmicity. Our findings suggest that the PXDLS motif plays an important role in circadian rhythmicity through regulation of protein interactions within the clock circuitry and that short linear motifs can be employed to modulate circadian oscillations.
昼夜节律核心时钟电路依赖于相互锁定的转录-翻译反馈环,这在很大程度上依赖于多种蛋白质相互作用。参与这些蛋白质复合物组装的分子机制相对未知。我们对涉及蛋白质相互作用的短线性基序进行的生物信息学分析表明,昼夜节律转录本中富含Pro-X-Asp-Leu-Ser(PXDLS)基序。我们发现PXDLS基序可以与BMAL1/CLOCK结合,并以细胞自主方式破坏昼夜节律振荡。值得注意的是,该基序在核心时钟蛋白REV-ERBα以及其他与时钟功能相关的蛋白质(NRIP1、CBP)中具有进化保守性。在此情况下,我们揭示了两个主要核心时钟反馈环之间的一种新型相互作用,并表明BMAL/CLOCK与REV-ERBα相互作用,且REV-ERBα的PXDLS基序参与它们的结合。此外,我们证明NRIP1和CBP的PXDLS基序参与昼夜节律性。我们的研究结果表明,PXDLS基序通过调节时钟电路内的蛋白质相互作用在昼夜节律性中发挥重要作用,并且短线性基序可用于调节昼夜节律振荡。