Neuromuscular and Regenerative Medicine Unit, School of Medical Sciences, The University of New South Wales, Sydney, New South Wales, Australia.
PLoS One. 2012;7(11):e49283. doi: 10.1371/journal.pone.0049283. Epub 2012 Nov 8.
GTF2IRD1 is one of the genes implicated in Williams-Beuren syndrome, a disease caused by haploinsufficiency of certain dosage-sensitive genes within a hemizygous microdeletion of chromosome 7. GTF2IRD1 is a prime candidate for some of the major features of the disease, presumably caused by abnormally reduced abundance of this putative transcriptional repressor protein. GTF2IRD1 has been shown to interact with the E3 SUMO ligase PIASxβ, but the significance of this relationship is largely unexplored. Here, we demonstrate that GTF2IRD1 can be SUMOylated by the SUMO E2 ligase UBC9 and the level of SUMOylation is enhanced by PIASxβ. A major SUMOylation site was mapped to lysine 495 within a conserved SUMO consensus motif. SUMOylation of GTF2IRD1 alters the affinity of the protein for binding partners that contain SUMO-interacting motifs, including a novel family member of the HDAC repressor complex, ZMYM5, and PIASxβ itself. In addition, we show that GTF2IRD1 is targeted for ubiquitination and proteasomal degradation. Cross regulation by SUMOylation modulates this process, thus potentially regulating the level of GTF2IRD1 protein in the cell. These findings, concerning post-translational control over the activity and stability of GTF2IRD1, together with previous work showing how GTF2IRD1 directly regulates its own transcription levels suggest an evolutionary requirement for fine control over GTF2IRD1 activity in the cell.
GTF2IRD1 是威廉姆斯-比伦综合征(Williams-Beuren syndrome)相关的基因之一,这种疾病是由于 7 号染色体半合子微缺失导致某些剂量敏感基因的杂合不足引起的。GTF2IRD1 是该疾病一些主要特征的主要候选基因,可能是由于这种假定的转录抑制蛋白的异常减少所致。已经表明 GTF2IRD1 可以与 E3 SUMO 连接酶 PIASxβ 相互作用,但这种关系的意义在很大程度上尚未得到探索。在这里,我们证明 GTF2IRD1 可以被 SUMO E2 连接酶 UBC9 进行 SUMO 化,并且 PIASxβ 增强了 SUMO 化的水平。一个主要的 SUMO 化位点被映射到保守的 SUMO 共有基序内的赖氨酸 495。GTF2IRD1 的 SUMO 化改变了该蛋白与包含 SUMO 相互作用基序的结合伴侣的亲和力,包括 HDAC 抑制剂复合物的一个新家族成员 ZMYM5 和 PIASxβ 本身。此外,我们表明 GTF2IRD1 是泛素化和蛋白酶体降解的靶标。SUMO 化的交叉调节调节了这个过程,从而可能调节细胞中 GTF2IRD1 蛋白的水平。这些发现涉及对 GTF2IRD1 的活性和稳定性的翻译后控制,以及之前的工作表明 GTF2IRD1 如何直接调节其自身的转录水平,这表明在细胞中对 GTF2IRD1 活性进行精细控制是进化所必需的。