Structural Basis for Recognition of Diubiquitins by NEMO

Yu Chih Lo, Su Chang Lin, Carla C. Rospigliosi, Dietrich B. Conze, Chuan Jin Wu, Jonathan D. Ashwell, David Eliezer, Hao Wu

Research output: Contribution to journalArticle

180 Citations (Scopus)

Abstract

NEMO is the regulatory subunit of the IκB kinase (IKK) in NF-κB activation, and its CC2-LZ region interacts with Lys63 (K63)-linked polyubiquitin to recruit IKK to receptor signaling complexes. In vitro, CC2-LZ also interacts with tandem diubiquitin. Here we report the crystal structure of CC2-LZ with two dimeric coiled coils representing CC2 and LZ, respectively. Surprisingly, mutagenesis and nuclear magnetic resonance experiments reveal that the binding sites for diubiquitins at LZ are composites of both chains and that each ubiquitin in diubiquitins interacts with symmetrical NEMO asymmetrically. For tandem diubiquitin, the first ubiquitin uses the conserved hydrophobic patch and the C-terminal tail, while the second ubiquitin uses an adjacent surface patch. For K63-linked diubiquitin, the proximal ubiquitin uses its conserved hydrophobic patch, while the distal ubiquitin mostly employs the C-terminal arm including the K63 linkage residue. These studies uncover the energetics and geometry for mutual recognition of NEMO and diubiquitins.

Original languageEnglish
Pages (from-to)602-615
Number of pages14
JournalMolecular Cell
Volume33
Issue number5
DOIs
Publication statusPublished - 2009 Mar 13

Fingerprint

Ubiquitin
Polyubiquitin
Mutagenesis
Phosphotransferases
Magnetic Resonance Spectroscopy
Binding Sites

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Cell Biology

Cite this

Lo, Y. C., Lin, S. C., Rospigliosi, C. C., Conze, D. B., Wu, C. J., Ashwell, J. D., ... Wu, H. (2009). Structural Basis for Recognition of Diubiquitins by NEMO. Molecular Cell, 33(5), 602-615. https://doi.org/10.1016/j.molcel.2009.01.012
Lo, Yu Chih ; Lin, Su Chang ; Rospigliosi, Carla C. ; Conze, Dietrich B. ; Wu, Chuan Jin ; Ashwell, Jonathan D. ; Eliezer, David ; Wu, Hao. / Structural Basis for Recognition of Diubiquitins by NEMO. In: Molecular Cell. 2009 ; Vol. 33, No. 5. pp. 602-615.
@article{691d74e812c146f5ae4a1f41410171a5,
title = "Structural Basis for Recognition of Diubiquitins by NEMO",
abstract = "NEMO is the regulatory subunit of the IκB kinase (IKK) in NF-κB activation, and its CC2-LZ region interacts with Lys63 (K63)-linked polyubiquitin to recruit IKK to receptor signaling complexes. In vitro, CC2-LZ also interacts with tandem diubiquitin. Here we report the crystal structure of CC2-LZ with two dimeric coiled coils representing CC2 and LZ, respectively. Surprisingly, mutagenesis and nuclear magnetic resonance experiments reveal that the binding sites for diubiquitins at LZ are composites of both chains and that each ubiquitin in diubiquitins interacts with symmetrical NEMO asymmetrically. For tandem diubiquitin, the first ubiquitin uses the conserved hydrophobic patch and the C-terminal tail, while the second ubiquitin uses an adjacent surface patch. For K63-linked diubiquitin, the proximal ubiquitin uses its conserved hydrophobic patch, while the distal ubiquitin mostly employs the C-terminal arm including the K63 linkage residue. These studies uncover the energetics and geometry for mutual recognition of NEMO and diubiquitins.",
author = "Lo, {Yu Chih} and Lin, {Su Chang} and Rospigliosi, {Carla C.} and Conze, {Dietrich B.} and Wu, {Chuan Jin} and Ashwell, {Jonathan D.} and David Eliezer and Hao Wu",
year = "2009",
month = "3",
day = "13",
doi = "10.1016/j.molcel.2009.01.012",
language = "English",
volume = "33",
pages = "602--615",
journal = "Molecular Cell",
issn = "1097-2765",
publisher = "Cell Press",
number = "5",

}

Lo, YC, Lin, SC, Rospigliosi, CC, Conze, DB, Wu, CJ, Ashwell, JD, Eliezer, D & Wu, H 2009, 'Structural Basis for Recognition of Diubiquitins by NEMO', Molecular Cell, vol. 33, no. 5, pp. 602-615. https://doi.org/10.1016/j.molcel.2009.01.012

Structural Basis for Recognition of Diubiquitins by NEMO. / Lo, Yu Chih; Lin, Su Chang; Rospigliosi, Carla C.; Conze, Dietrich B.; Wu, Chuan Jin; Ashwell, Jonathan D.; Eliezer, David; Wu, Hao.

In: Molecular Cell, Vol. 33, No. 5, 13.03.2009, p. 602-615.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Structural Basis for Recognition of Diubiquitins by NEMO

AU - Lo, Yu Chih

AU - Lin, Su Chang

AU - Rospigliosi, Carla C.

AU - Conze, Dietrich B.

AU - Wu, Chuan Jin

AU - Ashwell, Jonathan D.

AU - Eliezer, David

AU - Wu, Hao

PY - 2009/3/13

Y1 - 2009/3/13

N2 - NEMO is the regulatory subunit of the IκB kinase (IKK) in NF-κB activation, and its CC2-LZ region interacts with Lys63 (K63)-linked polyubiquitin to recruit IKK to receptor signaling complexes. In vitro, CC2-LZ also interacts with tandem diubiquitin. Here we report the crystal structure of CC2-LZ with two dimeric coiled coils representing CC2 and LZ, respectively. Surprisingly, mutagenesis and nuclear magnetic resonance experiments reveal that the binding sites for diubiquitins at LZ are composites of both chains and that each ubiquitin in diubiquitins interacts with symmetrical NEMO asymmetrically. For tandem diubiquitin, the first ubiquitin uses the conserved hydrophobic patch and the C-terminal tail, while the second ubiquitin uses an adjacent surface patch. For K63-linked diubiquitin, the proximal ubiquitin uses its conserved hydrophobic patch, while the distal ubiquitin mostly employs the C-terminal arm including the K63 linkage residue. These studies uncover the energetics and geometry for mutual recognition of NEMO and diubiquitins.

AB - NEMO is the regulatory subunit of the IκB kinase (IKK) in NF-κB activation, and its CC2-LZ region interacts with Lys63 (K63)-linked polyubiquitin to recruit IKK to receptor signaling complexes. In vitro, CC2-LZ also interacts with tandem diubiquitin. Here we report the crystal structure of CC2-LZ with two dimeric coiled coils representing CC2 and LZ, respectively. Surprisingly, mutagenesis and nuclear magnetic resonance experiments reveal that the binding sites for diubiquitins at LZ are composites of both chains and that each ubiquitin in diubiquitins interacts with symmetrical NEMO asymmetrically. For tandem diubiquitin, the first ubiquitin uses the conserved hydrophobic patch and the C-terminal tail, while the second ubiquitin uses an adjacent surface patch. For K63-linked diubiquitin, the proximal ubiquitin uses its conserved hydrophobic patch, while the distal ubiquitin mostly employs the C-terminal arm including the K63 linkage residue. These studies uncover the energetics and geometry for mutual recognition of NEMO and diubiquitins.

UR - http://www.scopus.com/inward/record.url?scp=61649103747&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=61649103747&partnerID=8YFLogxK

U2 - 10.1016/j.molcel.2009.01.012

DO - 10.1016/j.molcel.2009.01.012

M3 - Article

C2 - 19185524

AN - SCOPUS:61649103747

VL - 33

SP - 602

EP - 615

JO - Molecular Cell

JF - Molecular Cell

SN - 1097-2765

IS - 5

ER -

Lo YC, Lin SC, Rospigliosi CC, Conze DB, Wu CJ, Ashwell JD et al. Structural Basis for Recognition of Diubiquitins by NEMO. Molecular Cell. 2009 Mar 13;33(5):602-615. https://doi.org/10.1016/j.molcel.2009.01.012