Structural study of UFL1-UFC1 interaction uncovers the role of UFL1 N-terminal helix in ufmylation

Sayanika Banerjee; Julia K. Varga; Manoj Kumar; Guy Zoltsman; Shahar Rotem-Bamberger; Einav Cohen-Kfir; Michail N. Isupov; Rina Rosenzweig; Ora Schueler-Furman; Reuven Wiener

doi:10.15252/embr.202356920

Structural study of UFL1-UFC1 interaction uncovers the role of UFL1 N-terminal helix in ufmylation

Sayanika Banerjee, Julia K. Varga, Manoj Kumar, Guy Zoltsman, Shahar Rotem-Bamberger, Einav Cohen-Kfir, Michail N. Isupov, Rina Rosenzweig, Ora Schueler-Furman^*, Reuven Wiener^*

^*Corresponding author for this work

Department of Chemical and Structural Biology

Research output: Contribution to journal › Article › peer-review

5 Citations (Scopus)

Abstract

Ufmylation plays a crucial role in various cellular processes including DNA damage response, protein translation, and ER homeostasis. To date, little is known about how the enzymes responsible for ufmylation coordinate their action. Here, we study the details of UFL1 (E3) activity, its binding to UFC1 (E2), and its relation to UBA5 (E1), using a combination of structural modeling, X-ray crystallography, NMR, and biochemical assays. Guided by Alphafold2 models, we generate an active UFL1 fusion construct that includes its partner DDRGK1 and solve the crystal structure of this critical interaction. This fusion construct also unveiled the importance of the UFL1 N-terminal helix for binding to UFC1. The binding site suggested by our UFL1-UFC1 model reveals a conserved interface, and competition between UFL1 and UBA5 for binding to UFC1. This competition changes in the favor of UFL1 following UFM1 charging of UFC1. Altogether, our study reveals a novel, terminal helix-mediated regulatory mechanism, which coordinates the cascade of E1-E2-E3-mediated transfer of UFM1 to its substrate and provides new leads to target this modification.

Original language	English
Article number	e56920
Journal	EMBO Reports
Volume	24
Issue number	12
DOIs	https://doi.org/10.15252/embr.202356920
Publication status	Published Online - 21 Nov 2023

Funding

This work was supported, in whole or in part, by the Israel Science Foundation, founded by the Israel Academy of Science and Humanities (grant number 491/2021 to RW and grant number 301/2021 to OS‐F) and by the Israel Cancer Research Fund (award ID 21‐113‐PG to RW). JKV is supported by a Marie Sklodowska‐Curie European Training Network Grant #860517 (Ubimotif). We thank the staff of ESRF beamline ID30‐A for the help with data collection.

All Science Journal Classification (ASJC) codes

Biochemistry
Molecular Biology
Genetics

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title = "Structural study of UFL1-UFC1 interaction uncovers the role of UFL1 N-terminal helix in ufmylation",

abstract = "Ufmylation plays a crucial role in various cellular processes including DNA damage response, protein translation, and ER homeostasis. To date, little is known about how the enzymes responsible for ufmylation coordinate their action. Here, we study the details of UFL1 (E3) activity, its binding to UFC1 (E2), and its relation to UBA5 (E1), using a combination of structural modeling, X-ray crystallography, NMR, and biochemical assays. Guided by Alphafold2 models, we generate an active UFL1 fusion construct that includes its partner DDRGK1 and solve the crystal structure of this critical interaction. This fusion construct also unveiled the importance of the UFL1 N-terminal helix for binding to UFC1. The binding site suggested by our UFL1-UFC1 model reveals a conserved interface, and competition between UFL1 and UBA5 for binding to UFC1. This competition changes in the favor of UFL1 following UFM1 charging of UFC1. Altogether, our study reveals a novel, terminal helix-mediated regulatory mechanism, which coordinates the cascade of E1-E2-E3-mediated transfer of UFM1 to its substrate and provides new leads to target this modification.",

author = "Sayanika Banerjee and Varga, {Julia K.} and Manoj Kumar and Guy Zoltsman and Shahar Rotem-Bamberger and Einav Cohen-Kfir and Isupov, {Michail N.} and Rina Rosenzweig and Ora Schueler-Furman and Reuven Wiener",

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AU - Banerjee, Sayanika

AU - Varga, Julia K.

AU - Kumar, Manoj

AU - Zoltsman, Guy

AU - Rotem-Bamberger, Shahar

AU - Cohen-Kfir, Einav

AU - Isupov, Michail N.

AU - Rosenzweig, Rina

AU - Schueler-Furman, Ora

AU - Wiener, Reuven

PY - 2023/11/21

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Structural study of UFL1-UFC1 interaction uncovers the role of UFL1 N-terminal helix in ufmylation

Abstract

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