RT Journal Article
SR Electronic
T1 AlphaFold2-based fusion design deciphers crucial role of the E3 UFL1 N-terminal helix in E2 UFC1 binding and ufmylation
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 2022.09.15.508077
DO 10.1101/2022.09.15.508077
A1 Banerjee, Sayanika
A1 Varga, Julia K
A1 Kumar, Manoj
A1 Zoltsman, Guy
A1 Isupov, Michail N
A1 Rosenzweig, Rina
A1 Schueler-Furman, Ora
A1 Wiener, Reuven
YR 2022
UL http://biorxiv.org/content/early/2022/11/22/2022.09.15.508077.abstract
AB While protein modification by UFM1 (ufmylation) is highly appreciated as an important post-translational modification, little is known about the mechanisms of the enzymes responsible for this modification and in particular on the UFM1 E3 ligase, UFL1, that for its functionality has to form a complex with another protein DDRGK1 (UFBP1). Here we used AlphaFold2 to generate active, easily expressed, fusion proteins encompassing DDRGK1-UFL1. We then solved the crystal structure of this fusion, explaining the dependency of UFL1 on DDRGK1 to form a stable structure. In addition, we deciphered how UFL1, via its N-terminal helix, binds the E2, UFC1, and in turn, allows ufmylation. This mode of binding suggests a competition between E1 and E3 on E2 binding that is required for the proper transfer of UFM1 in the conjugation machinery.Competing Interest StatementThe authors have declared no competing interest.