Department of Cell and Developmental Biology, University of Colorado School of Medicine, Aurora, CO 80045-2537.
Linda Crnic Institute for Down Syndrome, University of Colorado School of Medicine, Aurora, CO 80045-2537.
Mol Biol Cell. 2022 Jul 1;33(8). doi: 10.1091/mbc.E21-10-0517-T. Epub 2022 Apr 27.
Trisomy 21, the source of Down syndrome, causes a 0.5-fold protein increase of the chromosome 21-resident gene Pericentrin (PCNT) and reduces primary cilia formation and signaling. We investigate how PCNT imbalances disrupt cilia. Using isogenic RPE-1 cells with increased chromosome 21 dosage, we find PCNT accumulates around the centrosome as a cluster of enlarged cytoplasmic puncta that localize along microtubules (MTs) and at MT ends. Cytoplasmic PCNT puncta impact the density, stability, and localization of the MT trafficking network required for primary cilia. The PCNT puncta appear to sequester cargo peripheral to centrosomes in what we call pericentrosomal crowding. The centriolar satellite proteins PCM1, CEP131, and CEP290, important for ciliogenesis, accumulate at enlarged PCNT puncta in trisomy 21 cells. Reducing PCNT when chromosome 21 ploidy is elevated is sufficient to decrease PCNT puncta and pericentrosomal crowding, reestablish a normal density of MTs around the centrosome, and restore ciliogenesis to wild-type levels. A transient reduction in MTs also decreases pericentrosomal crowding and partially rescues ciliogenesis in trisomy 21 cells, indicating that increased PCNT leads to defects in the MT network deleterious to normal centriolar satellite distribution. We propose that chromosome 21 aneuploidy disrupts MT-dependent intracellular trafficking required for primary cilia.
21 三体,唐氏综合征的根源,导致染色体 21 上驻留基因 Pericentrin(PCNT)的蛋白水平增加 0.5 倍,并减少初级纤毛的形成和信号转导。我们研究了 PCNT 失衡如何破坏纤毛。使用具有增加的 21 号染色体剂量的同基因 RPE-1 细胞,我们发现 PCNT 作为扩大的细胞质点状聚集体在中心体周围聚集,这些点状聚集体沿着微管(MTs)和 MT 末端定位。细胞质 PCNT 点状聚集体影响初级纤毛所需的 MT 运输网络的密度、稳定性和定位。这些 PCNT 点状聚集体似乎将位于中心体周围的货物隔离在我们所谓的中心体周围拥挤中。中心粒卫星蛋白 PCM1、CEP131 和 CEP290 对于纤毛发生很重要,在 21 三体细胞中在扩大的 PCNT 点状聚集体中积累。当 21 号染色体倍性升高时,减少 PCNT 足以减少 PCNT 点状聚集体和中心体周围拥挤,重建中心体周围 MT 的正常密度,并将纤毛发生恢复到野生型水平。MT 的短暂减少也会减少中心体周围的拥挤,并部分挽救 21 三体细胞中的纤毛发生,表明增加的 PCNT 导致 MT 网络缺陷,对正常中心粒卫星分布有害。我们提出,21 号染色体非整倍体破坏了初级纤毛所需的 MT 依赖性细胞内运输。