Molecular basis for arginine C-terminal degron recognition by Cul2FEM1 E3 ligase

Xinyan Chen; Shanhui Liao; Yaara Makaros; Qiong Guo; Zhongliang Zhu; Rina Krizelman; Karin Dahan; Xiaoming Tu; Xuebiao Yao; Itay Koren; Chao Xu

doi:10.1038/s41589-020-00704-3

Molecular basis for arginine C-terminal degron recognition by Cul2^FEM1 E3 ligase

Nat Chem Biol. 2021 Mar;17(3):254-262. doi: 10.1038/s41589-020-00704-3. Epub 2021 Jan 4.

Authors

Xinyan Chen^#¹, Shanhui Liao^#¹, Yaara Makaros^#², Qiong Guo¹, Zhongliang Zhu¹, Rina Krizelman², Karin Dahan², Xiaoming Tu¹, Xuebiao Yao¹, Itay Koren³, Chao Xu⁴

Affiliations

¹ MOE Key Laboratory for Membraneless Organelles and Cellular Dynamics, Hefei National Laboratory for Physical Sciences at the Microscale, School of Life Sciences, University of Science and Technology of China, Hefei, China.
² The Mina and Everard Goodman Faculty of Life Sciences, Bar-Ilan University, Ramat-Gan, Israel.
³ The Mina and Everard Goodman Faculty of Life Sciences, Bar-Ilan University, Ramat-Gan, Israel. itay.koren@biu.ac.il.
⁴ MOE Key Laboratory for Membraneless Organelles and Cellular Dynamics, Hefei National Laboratory for Physical Sciences at the Microscale, School of Life Sciences, University of Science and Technology of China, Hefei, China. xuchaor@ustc.edu.cn.

^# Contributed equally.

PMID: 33398168
DOI: 10.1038/s41589-020-00704-3

Abstract

Degrons are elements within protein substrates that mediate the interaction with specific degradation machineries to control proteolysis. Recently, a few classes of C-terminal degrons (C-degrons) that are recognized by dedicated cullin-RING ligases (CRLs) have been identified. Specifically, CRL2 using the related substrate adapters FEM1A/B/C was found to recognize C degrons ending with arginine (Arg/C-degron). Here, we uncover the molecular mechanism of Arg/C-degron recognition by solving a subset of structures of FEM1 proteins in complex with Arg/C-degron-bearing substrates. Our structural research, complemented by binding assays and global protein stability (GPS) analyses, demonstrates that FEM1A/C and FEM1B selectively target distinct classes of Arg/C-degrons. Overall, our study not only sheds light on the molecular mechanism underlying Arg/C-degron recognition for precise control of substrate turnover, but also provides valuable information for development of chemical probes for selectively regulating proteostasis.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Amino Acid Sequence
Arginine / chemistry*
Arginine / metabolism
Binding Sites
Carrier Proteins / chemistry*
Carrier Proteins / genetics
Carrier Proteins / metabolism
Cell Cycle Proteins / chemistry*
Cell Cycle Proteins / genetics
Cell Cycle Proteins / metabolism
Cloning, Molecular
Crystallography, X-Ray
Escherichia coli / genetics
Escherichia coli / metabolism
Gene Expression
Genetic Vectors / chemistry
Genetic Vectors / metabolism
HEK293 Cells
Humans
Models, Molecular
Proteasome Endopeptidase Complex / metabolism*
Protein Binding
Protein Conformation, alpha-Helical
Protein Conformation, beta-Strand
Protein Interaction Domains and Motifs
Proteolysis
Recombinant Proteins / chemistry
Recombinant Proteins / genetics
Recombinant Proteins / metabolism
Sequence Alignment
Sequence Homology, Amino Acid
Substrate Specificity
Ubiquitin-Protein Ligase Complexes / chemistry*
Ubiquitin-Protein Ligase Complexes / genetics
Ubiquitin-Protein Ligase Complexes / metabolism

Substances

Carrier Proteins
Cell Cycle Proteins
FEM1A protein, human
FEM1B protein, human
Recombinant Proteins
Arginine
Fem1c protein, human
Ubiquitin-Protein Ligase Complexes
Proteasome Endopeptidase Complex