ALKBH3 partner ASCC3 mediates P-body formation and selective clearance of MMS-induced 1-methyladenosine and 3-methylcytosine from mRNA

Journal of Translational Medicine

Hilde O. Erlandsen¹  Vuk Palibrk¹  Magnar Bjørås¹  Bjørnar Sporsheim²  Per Arne Aas³  Renana Rabe³  Tobias S. Iveland³  Kristian Lied Wollen³  Lars Hagen⁴  Davi M. Fonseca⁴  Geir Slupphaug⁴  Animesh Sharma⁴  Cathrine B. Vågbø⁴  Nima Mosammaparast⁵ 

[1] Department of Clinical and Molecular Medicine, Norwegian University of Science and Technology, NTNU, 7491, Trondheim, Norway;Department of Clinical and Molecular Medicine, Norwegian University of Science and Technology, NTNU, 7491, Trondheim, Norway;CMIC Cellular & Molecular Imaging Core Facility, Norwegian University of Science and Technology, NTNU, and the Central Norway Regional Health Authority Norway, Trondheim, Norway;Department of Clinical and Molecular Medicine, Norwegian University of Science and Technology, NTNU, 7491, Trondheim, Norway;Clinic of Laboratory Medicine, St. Olavs Hospital, Trondheim, Norway;Department of Clinical and Molecular Medicine, Norwegian University of Science and Technology, NTNU, 7491, Trondheim, Norway;Clinic of Laboratory Medicine, St. Olavs Hospital, Trondheim, Norway;PROMEC Core Facility for Proteomics and Modomics, Norwegian University of Science and Technology, NTNU, and the Central Norway Regional Health Authority Norway, Trondheim, Norway;Department of Pathology and Immunology, Division of Laboratory and Genomic Medicine, Washington University School of Medicine, 63110, St Louis, MO, USA;

关键词: Alkylating agents;    ALKBH3;    ASCC3;    Epitranscriptome;    1-Methyladenosine;    3-Methylcytosine;    7-Methylguanosine;    No-go decay;    P-bodies;    Ribosome quality control;   

DOI  :  10.1186/s12967-021-02948-6

来源: Springer

【 摘 要 】

BackgroundReversible enzymatic methylation of mammalian mRNA is widespread and serves crucial regulatory functions, but little is known to what degree chemical alkylators mediate overlapping modifications and whether cells distinguish aberrant from canonical methylations.MethodsHere we use quantitative mass spectrometry to determine the fate of chemically induced methylbases in the mRNA of human cells. Concomitant alteration in the mRNA binding proteome was analyzed by SILAC mass spectrometry.ResultsMMS induced prominent direct mRNA methylations that were chemically identical to endogenous methylbases. Transient loss of 40S ribosomal proteins from isolated mRNA suggests that aberrant methylbases mediate arrested translational initiation and potentially also no-go decay of the affected mRNA. Four proteins (ASCC3, YTHDC2, TRIM25 and GEMIN5) displayed increased mRNA binding after MMS treatment. ASCC3 is a binding partner of the DNA/RNA demethylase ALKBH3 and was recently shown to promote disassembly of collided ribosomes as part of the ribosome quality control (RQC) trigger complex. We find that ASCC3-deficient cells display delayed removal of MMS-induced 1-methyladenosine (m1A) and 3-methylcytosine (m3C) from mRNA and impaired formation of MMS-induced P-bodies.ConclusionsOur findings conform to a model in which ASCC3-mediated disassembly of collided ribosomes allows demethylation of aberrant m1A and m3C by ALKBH3. Our findings constitute first evidence of selective sanitation of aberrant mRNA methylbases over their endogenous counterparts and warrant further studies on RNA-mediated effects of chemical alkylators commonly used in the clinic.

【 授权许可】

CC BY