Uncovering the mechanisms of MuRF1-induced ubiquitylation and revealing similarities with MuRF2 and MuRF3

• DOI: 10.1016/j.bbrep.2023.101636
• Publication Date: 2024-01-06T00:14:56Z
• Authors: Samuel O. Lord; Peter W.J. Dawson; Jitpisute Chuntho...; Jimi Ng; Leslie M. Baehr; David C. Hughes; Pooja Sridhar; Timothy Knowles; Sue C. Bodine; Yu-Chiang Lai
• Keywords: Muscle Atrophy; 0301 basic medicine; Autoubiquitylation; Ubiquitin; QH301-705.5; RING E3 Ligase; QD415-436; Biochemistry; 03 medical and health sciences; In vitro; Ubiquitin Conjugating Enzyme (UBE2); Biology (General); Research Article

MuRF1 (Muscle-specific RING finger protein 1; gene name TRIM63) is a ubiquitin E3 ligase, associated with the progression of muscle atrophy. As a RING (Really Interesting New Gene) type E3 ligase, its unique activity of ubiquitylation is driven by a specific interaction with a UBE2 (ubiquitin conjugating enzyme). Our understanding of MuRF1 function remains unclear as candidate UBE2s have not been fully elucidated. In the present study, we screened human ubiquitin dependent UBE2s in vitro and found that MuRF1 engages in ubiquitylation with UBE2D, UBE2E, UBE2N/V families and UBE2W. MuRF1 can cause mono-ubiquitylation, K48- and K63-linked polyubiquitin chains in a UBE2 dependent manner. Moreover, we identified a two-step UBE2 dependent mechanism whereby MuRF1 is monoubiquitylated by UBE2W which acts as an anchor for UBE2N/V to generate polyubiquitin chains. With the in vitro ubiquitylation assay, we also found that MuRF2 and MuRF3 not only share the same UBE2 partners as MuRF1 but can also directly ubiquitylate the same substrates: Titin (A168-A170), Desmin, and MYLPF (Myosin Light Chain, Phosphorylatable, Fast Skeletal Muscle; also called Myosin Light Regulatory Chain 2). In summary, our work presents new insights into the mechanisms that underpin MuRF1 activity and reveals overlap in MuRF-induced ubiquitylation which could explain their partial redundancy in vivo.