UFMylation Modulates OFIP Stability and Centrosomal Localization

IF 2.6 4区 医学 Q2 MEDICAL LABORATORY TECHNOLOGY
Mulin Yang, Zihe Zhao, Jie Di, Dan Dong, Dengwen Li, Jie Ran
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引用次数: 0

Abstract

Background

OFIP, also known as KIAA0753, is a centrosomal and pericentriolar satellite protein implicated in ciliogenesis, centriolar duplication, and microtubule stability. In humans, genetic mutations affecting OFIP have been implicated in the pathogenesis of Oral-Facial-Digital (OFD) Syndrome and Joubert Syndrome. Ubiquitin-fold Modifier 1 (UFM1), the most recently identified ubiquitin-like protein, is covalently transferred to its substrates, in a process known as UFMylation. This modification has recently emerged as a key regulator of various biological processes by altering their stability, activity, or localization.

Methods

The interaction between UFL1 and OFIP, as well as the UFMylation of OFIP, were assessed through immunoprecipitation and immunoblotting analyses. The mRNA levels of OFIP were examined using reverse transcription quantitative PCR (RT-qPCR). Immunofluorescence microscopy was employed to examine the localization and distribution patterns of OFIP.

Results

Our findings demonstrate that UFL1 interacts with OFIP both in vivo and in vitro. We also found that OFIP undergoes UFMylation, and UFL1 promotes the OFIP UFMylation. Mechanistic studies demonstrate that OFIP UFMylation inhibits its protein stability and maintains its proper centrosomal localization. However, the efficacy of these regulatory mechanisms varies significantly between different cell types, being notably pronounced in HeLa cells but markedly reduced in RPE1 cells.

Conclusions

OFIP is identified as a novel substrate for UFMylation. UFL1-mediated OFIP UFMylation is essential for its stability and centrosomal localization in HeLa cells. However, these effects are not observed in RPE1 cells, highlighting cell type-specific heterogeneity in the role of OFIP UFMylation.

Abstract Image

ufmyation调节OFIP稳定性和中心体定位。
背景:OFIP,也被称为KIAA0753,是一种中心体和中心粒周围卫星蛋白,与纤毛发生、中心粒复制和微管稳定性有关。在人类中,影响OFIP的基因突变与口腔-面部-指(OFD)综合征和Joubert综合征的发病机制有关。泛素折叠修饰因子1 (UFM1)是最近发现的泛素样蛋白,在一个被称为ufmyation的过程中共价转移到其底物上。这种修饰最近通过改变生物过程的稳定性、活性或定位而成为各种生物过程的关键调节因子。方法:采用免疫沉淀法和免疫印迹法检测UFL1与OFIP的相互作用以及OFIP的ufmyation。采用反转录定量PCR (RT-qPCR)检测OFIP mRNA水平。采用免疫荧光显微镜检查OFIP的定位和分布模式。结果:我们的研究结果表明,UFL1在体内和体外都与OFIP相互作用。我们还发现OFIP经历了ufmyation,而UFL1促进了OFIP ufmyation。机制研究表明,OFIP ufmyation抑制其蛋白稳定性并维持其中心体的适当定位。然而,这些调节机制的功效在不同的细胞类型之间差异显著,在HeLa细胞中显著,而在RPE1细胞中显著降低。结论:OFIP被鉴定为ufmyation的一种新的底物。在HeLa细胞中,ufl1介导的OFIP ufmyation对于其稳定性和中心体定位至关重要。然而,在RPE1细胞中没有观察到这些作用,这突出了OFIP UFMylation作用的细胞类型特异性异质性。
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来源期刊
Journal of Clinical Laboratory Analysis
Journal of Clinical Laboratory Analysis 医学-医学实验技术
CiteScore
5.60
自引率
7.40%
发文量
584
审稿时长
6-12 weeks
期刊介绍: Journal of Clinical Laboratory Analysis publishes original articles on newly developing modes of technology and laboratory assays, with emphasis on their application in current and future clinical laboratory testing. This includes reports from the following fields: immunochemistry and toxicology, hematology and hematopathology, immunopathology, molecular diagnostics, microbiology, genetic testing, immunohematology, and clinical chemistry.
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