Martínez de Arrieta C, Morte B, Coloma A, Bernal J
Instituto de Investigaciones Biomédicas Alberto Sols, Consejo Superior de Investigaciones Cientificas, Universidad Autonoma de Madrid, Spain.
Endocrinology. 1999 Jan;140(1):335-43. doi: 10.1210/endo.140.1.6461.
NRGN is the human homolog of the neuron-specific rat RC3/neurogranin gene. This gene encodes a postsynaptic 78-amino acid protein kinase substrate that binds calmodulin in the absence of calcium, and that has been implicated in dendritic spine formation and synaptic plasticity. In the rat brain RC3 is under thyroid hormone control in specific neuronal subsets in both developing and adult animals. To evaluate whether the human gene is also a target of thyroid hormone we have searched for T3-responsive elements in NRGN cloned genomic fragments spanning the whole gene. Labeled DNA fragments were incubated with T3 receptors (T3R) and 9-cis-retinoic acid receptors and immunoprecipitated using an anti T3R antibody. A receptor-binding site was localized in the first intron, 3000 bp downstream from the origin of transcription. Footprinting analysis revealed the sequence GGATTAAATGAGGTAA, closely related to the consensus T3-responsive element of the direct repeat (DR4) type. This sequence binds the T3R-9-cis-retinoic acid receptors heterodimers, but not T3R monomers or homodimers, and is able to confer regulation by T3R and T3 when fused upstream of the NRGN or thymidine kinase promoters. The data reported in this work suggest that NRGN is a direct target of thyroid hormone in human brain, and that control of expression of this gene could underlay many of the consequences ofhypothyroidism on mental states during development as well as in adult subjects.
NRGN是神经元特异性大鼠RC3/神经颗粒素基因的人类同源物。该基因编码一种突触后78个氨基酸的蛋白激酶底物,在没有钙的情况下与钙调蛋白结合,并且与树突棘形成和突触可塑性有关。在大鼠脑中,RC3在发育中和成年动物的特定神经元亚群中受甲状腺激素控制。为了评估人类基因是否也是甲状腺激素的靶标,我们在跨越整个基因的NRGN克隆基因组片段中寻找T3反应元件。将标记的DNA片段与T3受体(T3R)和9-顺式视黄酸受体一起孵育,并用抗T3R抗体进行免疫沉淀。一个受体结合位点定位于第一个内含子,转录起始点下游3000 bp处。足迹分析揭示了序列GGATTAAATGAGGTAA,与直接重复(DR4)型的共有T3反应元件密切相关。该序列结合T3R-9-顺式视黄酸受体异二聚体,但不结合T3R单体或同二聚体,并且当在NRGN或胸苷激酶启动子上游融合时能够赋予T3R和T3调节作用。这项工作中报道的数据表明,NRGN是人类大脑中甲状腺激素的直接靶标,并且该基因表达的调控可能是甲状腺功能减退在发育过程以及成年个体中对精神状态产生许多影响的基础。