USP34 regulates tooth root morphogenesis by stabilizing NFIC

Abstract

Tooth root morphogenesis involves two biological processes, root elongation and dentinogenesis, which are guaranteed by downgrowth of Hertwig’s epithelial root sheath (HERS) and normal odontoblast differentiation. Ubiquitin-dependent protein degradation has been reported to precisely regulate various physiological processes, while its role in tooth development is still elusive. Here we show ubiquitin-specific protease 34 (USP34) plays a pivotal role in root formation. Deletion of Usp34 in dental mesenchymal cells leads to short root anomaly, characterized by truncated roots and thin root dentin. The USP34-deficient dental pulp cells (DPCs) exhibit decreased odontogenic differentiation with downregulation of nuclear factor I/C (NFIC). Overexpression of NFIC partially restores the impaired odontogenic potential of DPCs. These findings indicate that USP34-dependent deubiquitination is critical for root morphogenesis by stabilizing NFIC.

DNA demethylase ALKBH1 promotes adipogenic differentiation via regulation of HIF-1 signaling

“…We separated the protein by using 8% or 10% SDS–polyacrylamide gels and transferred it to PVDF membranes by a wet transfer apparatus (Bio-Rad). The membranes were blotted with 5% milk or bovine serum albumin (BSA) for 1 h and then incubated with primary antibodies at 4 °C overnight ( 70 ). The following antibodies were used: rabbit anti-ALKBH1 (1:2000, ab128895, Abcam), rabbit anti-HIF-1α (1:1000, ER1802-41, Huabio, Boston), rabbit anti-GYS1 (1:1000, ET1611-59, Huabio), rabbit anti-VEGF (1:2000, ET1604-28, Huabio), rabbit anti-GLUT1 (1:2000, ET1601-10, Huabio), rabbit anti-LDHA (1:2000, ET1608-57, Huabio), rabbit anti-CEBPA (1:2000, ET1612-46, Huabio), rabbit anti-FABP4 (1:2000, ET1703-98, Huabio), mouse anti-α-tubulin (1:2,000, sc-32293; Santa Cruz Biotechnology).…”

“…We separated the protein by using 8% or 10% SDS-polyacrylamide gels and transferred it to PVDF membranes by a wet transfer apparatus (Bio-Rad). The membranes were blotted with 5% milk or bovine serum albumin (BSA) for 1 h and then incubated with primary antibodies at 4℃ overnight [70]. The following antibodies were used: rabbit anti-ALKBH1 (1:2000, ab128895, Abcam, Cambridge, MA), rabbit anti-HIF-1α…”

NEDD4 E3 Ligases: Functions and Mechanisms in Bone and Tooth

“…Deubiquitination can reverse the process of ubiquitination, by which deubiquitinases remove ubiquitin from ubiquitin-modified proteins [ 141 ]. Conditional deletion of USP34 in the dental mesenchyme cells disturbed deubiquitination-mediated NFIC stability, thus frustrating the odontogenic potential of DPCs, and a short root anomaly was observed in USP34 deletion mice [ 142 ]. USP49 was newly identified as a deubiquitinating enzyme of two transcription factors responsible for hypodontia and oligodontia, paired box gene 9 (PAX9) and Msh Homeobox 1 (MSX1), to stabilize their protein levels.…”

Ubiquitination and Deubiquitination in Oral Disease

“…The numerous possible causes for tooth sensitivity include worn tooth enamel and exposed tooth root surface, among others. Shuang Jiang et al reported the USP34-deficient dental pulp cells (DPCs) exhibit decreased odontogenic differentiation with downregulation of nuclear factor I/C (NFIC) and Overexpression of NFIC partially restores the impaired odontogenic potential of DPCs [ 107 ]. They reported that ubiquitin-specific protease 34 (USP34) plays a pivotal role in tooth root formation; findings indicate that USP34-dependent deubiquitination is critical for root morphogenesis by stabilizing NFIC [ 107 ].…”