Jensvold Zena D, Christenson Anna E, Flood Julia R, Lewis Peter W
Department of Biomolecular Chemistry, School of Medicine and Public Health, University of Wisconsin, Madison, WI 53715, USA.
bioRxiv. 2024 Feb 1:2023.12.28.573526. doi: 10.1101/2023.12.28.573526.
The Human Silencing Hub (HuSH) complex is composed of TASOR, MPP8, and PPHLN1 subunits and serves as a conserved protein complex responsible for silencing transposable elements in vertebrate animals. Despite its importance, the regulatory mechanisms and recruitment dynamics governing this complex remain poorly understood. In this study, we have identified a second HuSH complex, termed HuSH2, centered around TASOR2, a paralog of the core TASOR protein in HuSH. Our findings indicate that every subunit in both HuSH and HuSH2 has an important role in achieving precise genomic localization to distinct, non-overlapping genomic loci. We utilized in silico protein structure prediction to simulate the interactions between MPP8 and both TASOR paralogs. Drawing on the insights gained from these predictions, we implemented amino acid substitutions that interfered with the binding of MPP8 to each HuSH complex. Leveraging these MPP8 transgenes and other constructs, we identified an important role played by the relative quantities of HuSH complexes in controlling the activity of LINE-1 elements. Furthermore, our results suggest that dynamic changes in TASOR and TASOR2 expression enable cells to finely tune the extent of HuSH-mediated silencing. Our study provides insights into the intricate interplay between HuSH complexes, illuminating their important role in the regulation of retrotransposon silencing.
人类沉默中心(HuSH)复合体由TASOR、MPP8和PPHLN1亚基组成,是一种保守的蛋白质复合体,负责沉默脊椎动物中的转座元件。尽管其很重要,但调控该复合体的机制和募集动态仍知之甚少。在本研究中,我们鉴定出了第二种HuSH复合体,称为HuSH2,它以TASOR2为核心,TASOR2是HuSH中核心TASOR蛋白的旁系同源物。我们的研究结果表明,HuSH和HuSH2中的每个亚基在实现精确基因组定位到不同的、不重叠的基因组位点方面都发挥着重要作用。我们利用计算机蛋白质结构预测来模拟MPP8与两种TASOR旁系同源物之间的相互作用。基于这些预测获得的见解,我们进行了氨基酸替换,干扰了MPP8与每个HuSH复合体的结合。利用这些MPP8转基因和其他构建体,我们确定了HuSH复合体的相对数量在控制LINE-1元件活性方面发挥的重要作用。此外,我们的结果表明,TASOR和TASOR2表达的动态变化使细胞能够精细调节HuSH介导的沉默程度。我们的研究深入了解了HuSH复合体之间的复杂相互作用,阐明了它们在逆转录转座子沉默调控中的重要作用。