Exploration of the TRIM fold of MuRF1 using EPR reveals a canonical antiparallel structure and extended COS-box

Michael Stevens, Barbara Franke, Katarzyna A. Skorupka, David S. Cafiso, Owen Pornillos, Olga Mayans, David Norman (Lead / Corresponding author)

Research output: Contribution to journalArticle

1 Citation (Scopus)
53 Downloads (Pure)

Abstract

MuRF1 (TRIM63) is a RING-type E3 ubiquitin ligase with a predicted tripartite TRIM fold. TRIM proteins rely upon the correct placement of an N-terminal RING domain, with respect to C-terminal, specific substrate-binding domains. The TRIM domain organization is orchestrated by a central helical domain that forms an antiparallel coiled-coil motif and mediates the dimerization of the fold. MuRF1 has a reduced TRIM composition characterized by a lack of specific substrate binding domains, but contains in its helical domain a conserved sequence motif termed COS-box that has been speculated to fold independently into an α-hairpin. These characteristics had led to question whether MuRF1 adopts a canonical TRIM fold. Using a combination of electron paramagnetic resonance, on spin-labeled protein, and disulfide crosslinking, we show that TRIM63 follows the structural conservation of the TRIM dimerization domain, observed in other proteins. We also show that the COS-box motif folds back onto the dimerization coiled-coil motif, predictably forming a four-helical bundle at the center of the protein and emulating the architecture of canonical TRIMs.

Original languageEnglish
Pages (from-to)2900-2909
Number of pages10
JournalJournal of Molecular Biology
Volume431
Issue number15
Early online date22 May 2019
DOIs
Publication statusPublished - 12 Jul 2019

Fingerprint

Dimerization
Proteins
Ubiquitin-Protein Ligases
Conserved Sequence
Electron Spin Resonance Spectroscopy
Disulfides

Keywords

  • MuRF1
  • PELDOR
  • TRIM fold
  • coiled-coil
  • disulfide cross-linking

Cite this

Stevens, Michael ; Franke, Barbara ; Skorupka, Katarzyna A. ; Cafiso, David S. ; Pornillos, Owen ; Mayans, Olga ; Norman, David. / Exploration of the TRIM fold of MuRF1 using EPR reveals a canonical antiparallel structure and extended COS-box. In: Journal of Molecular Biology. 2019 ; Vol. 431, No. 15. pp. 2900-2909.
@article{c3f3ade0fb8743fc8f77715f2a5a81b3,
title = "Exploration of the TRIM fold of MuRF1 using EPR reveals a canonical antiparallel structure and extended COS-box",
abstract = "MuRF1 (TRIM63) is a RING-type E3 ubiquitin ligase with a predicted tripartite TRIM fold. TRIM proteins rely upon the correct placement of an N-terminal RING domain, with respect to C-terminal, specific substrate-binding domains. The TRIM domain organization is orchestrated by a central helical domain that forms an antiparallel coiled-coil motif and mediates the dimerization of the fold. MuRF1 has a reduced TRIM composition characterized by a lack of specific substrate binding domains, but contains in its helical domain a conserved sequence motif termed COS-box that has been speculated to fold independently into an α-hairpin. These characteristics had led to question whether MuRF1 adopts a canonical TRIM fold. Using a combination of electron paramagnetic resonance, on spin-labeled protein, and disulfide crosslinking, we show that TRIM63 follows the structural conservation of the TRIM dimerization domain, observed in other proteins. We also show that the COS-box motif folds back onto the dimerization coiled-coil motif, predictably forming a four-helical bundle at the center of the protein and emulating the architecture of canonical TRIMs.",
keywords = "MuRF1, PELDOR, TRIM fold, coiled-coil, disulfide cross-linking",
author = "Michael Stevens and Barbara Franke and Skorupka, {Katarzyna A.} and Cafiso, {David S.} and Owen Pornillos and Olga Mayans and David Norman",
note = "This research was supported by DFG SFB969, the Leducq Foundation (TNE-13CVD04), The Wellcome Trust 099149/Z/12/Z, U.S. National Institutes of Health grants R01-GM035215 and R01-GM112508, and the IMR CDT",
year = "2019",
month = "7",
day = "12",
doi = "10.1016/j.jmb.2019.05.025",
language = "English",
volume = "431",
pages = "2900--2909",
journal = "Journal of Molecular Biology",
issn = "0022-2836",
publisher = "Elsevier",
number = "15",

}

Exploration of the TRIM fold of MuRF1 using EPR reveals a canonical antiparallel structure and extended COS-box. / Stevens, Michael; Franke, Barbara ; Skorupka, Katarzyna A.; Cafiso, David S. ; Pornillos, Owen ; Mayans, Olga ; Norman, David (Lead / Corresponding author).

In: Journal of Molecular Biology, Vol. 431, No. 15, 12.07.2019, p. 2900-2909.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Exploration of the TRIM fold of MuRF1 using EPR reveals a canonical antiparallel structure and extended COS-box

AU - Stevens, Michael

AU - Franke, Barbara

AU - Skorupka, Katarzyna A.

AU - Cafiso, David S.

AU - Pornillos, Owen

AU - Mayans, Olga

AU - Norman, David

N1 - This research was supported by DFG SFB969, the Leducq Foundation (TNE-13CVD04), The Wellcome Trust 099149/Z/12/Z, U.S. National Institutes of Health grants R01-GM035215 and R01-GM112508, and the IMR CDT

PY - 2019/7/12

Y1 - 2019/7/12

N2 - MuRF1 (TRIM63) is a RING-type E3 ubiquitin ligase with a predicted tripartite TRIM fold. TRIM proteins rely upon the correct placement of an N-terminal RING domain, with respect to C-terminal, specific substrate-binding domains. The TRIM domain organization is orchestrated by a central helical domain that forms an antiparallel coiled-coil motif and mediates the dimerization of the fold. MuRF1 has a reduced TRIM composition characterized by a lack of specific substrate binding domains, but contains in its helical domain a conserved sequence motif termed COS-box that has been speculated to fold independently into an α-hairpin. These characteristics had led to question whether MuRF1 adopts a canonical TRIM fold. Using a combination of electron paramagnetic resonance, on spin-labeled protein, and disulfide crosslinking, we show that TRIM63 follows the structural conservation of the TRIM dimerization domain, observed in other proteins. We also show that the COS-box motif folds back onto the dimerization coiled-coil motif, predictably forming a four-helical bundle at the center of the protein and emulating the architecture of canonical TRIMs.

AB - MuRF1 (TRIM63) is a RING-type E3 ubiquitin ligase with a predicted tripartite TRIM fold. TRIM proteins rely upon the correct placement of an N-terminal RING domain, with respect to C-terminal, specific substrate-binding domains. The TRIM domain organization is orchestrated by a central helical domain that forms an antiparallel coiled-coil motif and mediates the dimerization of the fold. MuRF1 has a reduced TRIM composition characterized by a lack of specific substrate binding domains, but contains in its helical domain a conserved sequence motif termed COS-box that has been speculated to fold independently into an α-hairpin. These characteristics had led to question whether MuRF1 adopts a canonical TRIM fold. Using a combination of electron paramagnetic resonance, on spin-labeled protein, and disulfide crosslinking, we show that TRIM63 follows the structural conservation of the TRIM dimerization domain, observed in other proteins. We also show that the COS-box motif folds back onto the dimerization coiled-coil motif, predictably forming a four-helical bundle at the center of the protein and emulating the architecture of canonical TRIMs.

KW - MuRF1

KW - PELDOR

KW - TRIM fold

KW - coiled-coil

KW - disulfide cross-linking

UR - http://www.scopus.com/inward/record.url?scp=85066798666&partnerID=8YFLogxK

U2 - 10.1016/j.jmb.2019.05.025

DO - 10.1016/j.jmb.2019.05.025

M3 - Article

C2 - 31125568

VL - 431

SP - 2900

EP - 2909

JO - Journal of Molecular Biology

JF - Journal of Molecular Biology

SN - 0022-2836

IS - 15

ER -