K63-Ubiquitylation and TRAF6 Pathways Regulate Mammalian P-Body Formation and mRNA Decapping.
Signals and posttranslational modifications regulating the decapping step in mRNA degradation pathways are poorly defined. In this study we reveal the importance of K63-linked ubiquitylation for the assembly of decapping factors, P-body formation, and constitutive decay of instable mRNAs encoding mediators of inflammation by various experimental approaches. K63-branched ubiquitin chains also regulate IL-1-inducible phosphorylation of the P-body component DCP1a. The E3 ligase TRAF6 binds to DCP1a and indirectly regulates DCP1a phosphorylation, expression of decapping factors, and gene-specific mRNA decay. Mutation of six C-terminal lysines of DCP1a suppresses decapping activity and impairs the interaction with the mRNA decay factors DCP2, EDC4, and XRN1, but not EDC3, thus remodeling P-body architecture. The usage of ubiquitin chains for the proper assembly and function of the decay-competent mammalian decapping complex suggests an additional layer of control to allow a coordinated function of decapping activities and mRNA metabolism in higher eukaryotes.
Rudolf-Buchheim-Institute of Pharmacology, Justus-Liebig-University Giessen, 35392 Giessen, Germany.
Ulas
U
Tenekeci
Rudolf-Buchheim-Institute of Pharmacology, Justus-Liebig-University Giessen, 35392 Giessen, Germany.
Michael
M
Poppe
Rudolf-Buchheim-Institute of Pharmacology, Justus-Liebig-University Giessen, 35392 Giessen, Germany.
Knut
K
Beuerlein
Rudolf-Buchheim-Institute of Pharmacology, Justus-Liebig-University Giessen, 35392 Giessen, Germany.
Christin
C
Buro
Rudolf-Buchheim-Institute of Pharmacology, Justus-Liebig-University Giessen, 35392 Giessen, Germany.
Helmut
H
Müller
Rudolf-Buchheim-Institute of Pharmacology, Justus-Liebig-University Giessen, 35392 Giessen, Germany.
Hendrik
H
Weiser
Rudolf-Buchheim-Institute of Pharmacology, Justus-Liebig-University Giessen, 35392 Giessen, Germany.
Daniela
D
Kettner-Buhrow
Rudolf-Buchheim-Institute of Pharmacology, Justus-Liebig-University Giessen, 35392 Giessen, Germany.
Katharina
K
Porada
Rudolf-Buchheim-Institute of Pharmacology, Justus-Liebig-University Giessen, 35392 Giessen, Germany.
Doris
D
Newel
Department of Molecular Biology, Howard Hughes Medical Institute, University of Texas Southwestern Medical Center, Dallas, TX 75390-9148, USA.
Ming
M
Xu
Department of Molecular Biology, Howard Hughes Medical Institute, University of Texas Southwestern Medical Center, Dallas, TX 75390-9148, USA.
Zhijian J
ZJ
Chen
Institute of Biochemistry, Justus-Liebig-University Giessen, 35392 Giessen, Germany.
Julia
J
Busch
Institute of Biochemistry, Justus-Liebig-University Giessen, 35392 Giessen, Germany.
M Lienhard
ML
Schmitz
Rudolf-Buchheim-Institute of Pharmacology, Justus-Liebig-University Giessen, 35392 Giessen, Germany. Electronic address: michael.kracht@pharma.med.uni-giessen.de.
Michael
M
Kracht
D000818:Animals; D045744:Cell Line, Tumor; D004722:Endoribonucleases / Q000235:genetics / Q000378:metabolism*; D005095:Exoribonucleases / Q000378:metabolism; D057809:HEK293 Cells; D006801:Humans; D053582:Interleukin-1alpha / Q000494:pharmacology; D047908:Intracellular Signaling Peptides and Proteins; D008239:Lysine / Q000378:metabolism*; D051379:Mice; D008869:Microtubule-Associated Proteins / Q000378:metabolism; D009154:Mutation; D010766:Phosphorylation; D011485:Protein Binding; D054730:Protein Interaction Domains and Motifs; D011506:Proteins / Q000378:metabolism; D012315:RNA Caps / Q000235:genetics / Q000378:metabolism*; D020871:RNA Stability / Q000187:drug effects; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism*; D017472:Receptors, Interleukin-1 / Q000819:agonists / Q000235:genetics / Q000378:metabolism; D048029:TNF Receptor-Associated Factor 6 / Q000235:genetics / Q000378:metabolism*; D013997:Time Factors; D015534:Trans-Activators / Q000235:genetics / Q000378:metabolism*; D014162:Transfection; D054875:Ubiquitination / Q000187:drug effects
C507914:EDC4 protein, human; C508584:IL1A protein, human; D053582:Interleukin-1alpha; D047908:Intracellular Signaling Peptides and Proteins; D008869:Microtubule-Associated Proteins; D011506:Proteins; D012315:RNA Caps; D012333:RNA, Messenger; D017472:Receptors, Interleukin-1; D048029:TNF Receptor-Associated Factor 6; C000604754:Tifab protein, human; D015534:Trans-Activators; D004722:Endoribonucleases; D005095:Exoribonucleases; C470512:XRN1 protein, human; C466279:DCP1A protein, human; C467723:DCP2 protein, human; D008239:Lysine
DCP1a; IL-1; K63R ubiquitin; P-body; TRAF6; ubiquitin