Department of Neurology, University of Michigan, Ann Arbor, MI, 48109-2200, USA.
Graduate Program in Cellular and Molecular Biology, University of Michigan, Ann Arbor, MI, 48109-2200, USA.
Cell Mol Life Sci. 2022 Mar 5;79(3):176. doi: 10.1007/s00018-022-04170-z.
The brain-expressed ubiquilins (UBQLNs) 1, 2 and 4 are a family of ubiquitin adaptor proteins that participate broadly in protein quality control (PQC) pathways, including the ubiquitin proteasome system (UPS). One family member, UBQLN2, has been implicated in numerous neurodegenerative diseases including ALS/FTD. UBQLN2 typically resides in the cytoplasm but in disease can translocate to the nucleus, as in Huntington's disease where it promotes the clearance of mutant Huntingtin. How UBQLN2 translocates to the nucleus and clears aberrant nuclear proteins, however, is not well understood. In a mass spectrometry screen to discover UBQLN2 interactors, we identified a family of small (13 kDa), highly homologous uncharacterized proteins, RTL8, and confirmed the interaction between UBQLN2 and RTL8 both in vitro using recombinant proteins and in vivo using mouse brain tissue. Under endogenous and overexpressed conditions, RTL8 localizes to nucleoli. When co-expressed with UBQLN2, RTL8 promotes nuclear translocation of UBQLN2. RTL8 also facilitates UBQLN2's nuclear translocation during heat shock. UBQLN2 and RTL8 colocalize within ubiquitin-enriched subnuclear structures containing PQC components. The robust effect of RTL8 on the nuclear translocation and subnuclear localization of UBQLN2 does not extend to the other brain-expressed ubiquilins, UBQLN1 and UBQLN4. Moreover, compared to UBQLN1 and UBQLN4, UBQLN2 preferentially stabilizes RTL8 levels in human cell lines and in mouse brain, supporting functional heterogeneity among UBQLNs. As a novel UBQLN2 interactor that recruits UBQLN2 to specific nuclear compartments, RTL8 may regulate UBQLN2 function in nuclear protein quality control.
脑表达的泛素结合蛋白(UBQLNs)1、2 和 4 是一类泛素衔接蛋白,广泛参与蛋白质量控制(PQC)途径,包括泛素蛋白酶体系统(UPS)。一个家族成员 UBQLN2 与包括 ALS/FTD 在内的多种神经退行性疾病有关。UBQLN2 通常位于细胞质中,但在疾病中可以易位到细胞核,如在亨廷顿病中,它促进突变亨廷顿蛋白的清除。然而,UBQLN2 如何易位到细胞核并清除异常核蛋白尚不清楚。在一项用于发现 UBQLN2 相互作用蛋白的质谱筛选中,我们鉴定了一组小(13 kDa)、高度同源的未鉴定蛋白 RTL8,并使用重组蛋白在体外和使用小鼠脑组织在体内证实了 UBQLN2 与 RTL8 之间的相互作用。在内源性和过表达条件下,RTL8 定位于核仁。当与 UBQLN2 共表达时,RTL8 促进 UBQLN2 的核易位。RTL8 还在热休克期间促进 UBQLN2 的核易位。UBQLN2 和 RTL8 在含有 PQC 成分的富含泛素的亚核结构内共定位。RTL8 对 UBQLN2 的核易位和亚核定位的强烈影响不适用于其他脑表达的泛素结合蛋白 UBQLN1 和 UBQLN4。此外,与 UBQLN1 和 UBQLN4 相比,UBQLN2 更倾向于在人细胞系和小鼠脑中稳定 RTL8 水平,支持 UBQLNs 之间的功能异质性。作为一种新的 UBQLN2 相互作用蛋白,它将 UBQLN2 招募到特定的核区室,RTL8 可能调节 UBQLN2 在核蛋白质量控制中的功能。