The kinase GSK-3 alters the repertoire of RNA binding proteins present on lipid metabolism transcripts leading to altered expression in the C. elegans nervous system

Tissue-specific regulation of gene expression is essential for multicellular organisms, and RNA binding proteins play central roles in these molecular processes. To determine how the Caenorhabditis elegans RNA binding protein ADR-1 regulates tissue-specific gene expression, we profiled the binding targets of ADR-1 in neural cells and assessed the effects of ADR-1 RNA binding on neural gene expression. We identified a cohort of neural transcripts that facilitate lipid synthesis and are directly regulated by ADR-1 RNA binding. To identify cellular factors that influence ADR-1 binding, a forward genetic screen was performed, revealing that the serine/threonine protein kinase, glycogen synthase kinase-3 (GSK-3), inhibits ADR-1 binding to the cohort. Further investigation revealed that RNA binding protein VIG-1, promotes ADR-1 binding to the cohort in a GSK-3-dependent manner. Together, we reveal that interplay between kinases and RNA binding proteins regulates expression of lipid metabolism genes within neural cells, potentially impacting stress resistance and longevity. High throughput sequencing of neural ADR-1 immunoprecipitated samples and lysates in the presence and absence of adr-2; high throughput sequencing of neural cells isolated from adr-2(-) and adr-2(-);dsRBD1 mutant animals

基因表达的组织特异性调控对于多细胞生物而言至关重要，RNA结合蛋白(RNA binding protein)在这类分子进程中扮演核心角色。为明确秀丽隐杆线虫（Caenorhabditis elegans）RNA结合蛋白ADR-1如何调控组织特异性基因表达，我们对神经细胞内ADR-1的结合靶标开展了谱学分析，并评估了ADR-1的RNA结合活性对神经基因表达的影响。我们鉴定出一组可促进脂质合成且受ADR-1 RNA结合直接调控的神经转录本。为筛选影响ADR-1结合活性的细胞因子，我们实施了正向遗传筛选，结果发现丝氨酸/苏氨酸蛋白激酶糖原合成激酶3（glycogen synthase kinase-3, GSK-3）会抑制ADR-1与该组转录本的结合。进一步研究显示，RNA结合蛋白VIG-1可通过依赖GSK-3的方式促进ADR-1与该组转录本的结合。综上，我们揭示了蛋白激酶与RNA结合蛋白之间的相互作用可调控神经细胞内脂质代谢基因的表达，该调控过程或可影响生物的抗逆能力与寿命。对存在adr-2与缺失adr-2条件下的神经ADR-1免疫沉淀样本及裂解液进行高通量测序；对从adr-2(-)及adr-2(-);dsRBD1突变体动物中分离得到的神经细胞开展高通量测序。