title: Muscle-specific RING finger 1 is a bona fide ubiquitin ligase that degrades cardiac troponin I.
abstract: Muscle-specific RING finger protein 1 (MuRF1) is a sarcomere-associated protein that is restricted to cardiac and skeletal muscle. In skeletal muscle, MuRF1 is up-regulated by conditions that provoke atrophy, but its function in the heart is not known. The presence of a RING finger in MuRF1 raises the possibility that it is a component of the ubiquitin-proteasome system of protein degradation. We performed a yeast two-hybrid screen to search for interaction partners of MuRF1 in the heart that might be targets of its putative ubiquitin ligase activity. This screen identified troponin I as a MuRF1 partner protein. MuRF1 and troponin I were found to associate both in vitro and in vivo in cultured cardiomyocytes. MuRF1 reduced steady-state troponin I levels when coexpressed in COS-7 cells and increased degradation of endogenous troponin I protein in cardiomyocytes. The degradation of troponin I in cardiomyocytes was associated with the accumulation of ubiquitylated intermediates of troponin I and was proteasome-dependent. In vitro, MuRF1 functioned as a ubiquitin ligase to catalyze ubiquitylation of troponin I through a RING finger-dependent mechanism. In isolated cardiomyocytes, MuRF1 reduced indices of contractility. In cardiomyocytes, these processes may determine the balance between hypertrophic and antihypertrophic signals and the regulation of myocyte contractile responses in the setting of heart failure.
authors.affiliation: Carolina Cardiovascular Biology Center, University of North Carolina, Chapel Hill, NC 27599-7126, USA.
authors.forename: Vishram
authors.initials: V
authors.lastname: Kedar
authors.forename: Holly
authors.initials: H
authors.lastname: McDonough
authors.forename: Ranjana
authors.initials: R
authors.lastname: Arya
authors.forename: Hui-Hua
authors.initials: HH
authors.lastname: Li
authors.forename: Howard A
authors.initials: HA
authors.lastname: Rockman
authors.forename: Cam
authors.initials: C
authors.lastname: Patterson
mesh_terms: D000818:Animals; D000831:Animals, Newborn; D001284:Atrophy; D019556:COS Cells; D002384:Catalysis; D002478:Cells, Cultured; D019084:Fluorescent Antibody Technique, Indirect; D015723:Gene Library; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006321:Heart / Q000502:physiology; D006352:Heart Ventricles / Q000166:cytology; D006801:Humans; D006984:Hypertrophy / Q000473:pathology; D009124:Muscle Proteins; D009206:Myocardium / Q000378:metabolism; D010957:Plasmids / Q000378:metabolism; D046988:Proteasome Endopeptidase Complex / Q000378:metabolism; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D051381:Rats; D013997:Time Factors; D000070584:Tripartite Motif Proteins; D019210:Troponin I / Q000737:chemistry / Q000378:metabolism*; D020798:Two-Hybrid System Techniques; D025801:Ubiquitin / Q000378:metabolism; D044767:Ubiquitin-Protein Ligases / Q000737:chemistry* / Q000378:metabolism / Q000502:physiology*; D015854:Up-Regulation
chemical_list: D009124:Muscle Proteins; D000070584:Tripartite Motif Proteins; D019210:Troponin I; D025801:Ubiquitin; D049452:Green Fluorescent Proteins; C440908:TRIM63 protein, human; C499086:Trim63 protein, rat; D044767:Ubiquitin-Protein Ligases; D046988:Proteasome Endopeptidase Complex