Dual RING E3 Architectures Regulate Multiubiquitination and Ubiquitin Chain Elongation by APC/C.
Protein ubiquitination involves E1, E2, and E3 trienzyme cascades. E2 and RING E3 enzymes often collaborate to first prime a substrate with a single ubiquitin (UB) and then achieve different forms of polyubiquitination: multiubiquitination of several sites and elongation of linkage-specific UB chains. Here, cryo-EM and biochemistry show that the human E3 anaphase-promoting complex/cyclosome (APC/C) and its two partner E2s, UBE2C (aka UBCH10) and UBE2S, adopt specialized catalytic architectures for these two distinct forms of polyubiquitination. The APC/C RING constrains UBE2C proximal to a substrate and simultaneously binds a substrate-linked UB to drive processive multiubiquitination. Alternatively, during UB chain elongation, the RING does not bind UBE2S but rather lures an evolving substrate-linked UB to UBE2S positioned through a cullin interaction to generate a Lys11-linked chain. Our findings define mechanisms of APC/C regulation, and establish principles by which specialized E3-E2-substrate-UB architectures control different forms of polyubiquitination.
Department of Structural Biology, St. Jude Children's Research Hospital, Memphis, TN 38105, USA.
Nicholas G
NG
Brown
Department of Structural Biology, St. Jude Children's Research Hospital, Memphis, TN 38105, USA; Howard Hughes Medical Institute, Memphis, TN 38105, USA.
Ryan
R
VanderLinden
Department of Structural Biology, St. Jude Children's Research Hospital, Memphis, TN 38105, USA.
Edmond R
ER
Watson
Research Institute of Molecular Pathology (IMP), Vienna Biocenter (VBC), 1030 Vienna, Austria.
Florian
F
Weissmann
Department of Cell Biology, Harvard Medical School, Boston, MA 02115, USA.
Alban
A
Ordureau
Department of Structural Biology, St. Jude Children's Research Hospital, Memphis, TN 38105, USA.
Kuen-Phon
KP
Wu
Donnelly Centre for Cellular and Biomolecular Research, University of Toronto, 160 College Street, Toronto, Ontario M5S3E1, Canada.
Wei
W
Zhang
Department of Structural Biology, St. Jude Children's Research Hospital, Memphis, TN 38105, USA.
Shanshan
S
Yu
Department of Structural Biology, St. Jude Children's Research Hospital, Memphis, TN 38105, USA.
Peter Y
PY
Mercredi
Department of Biochemistry and Biophysics and Lineberger Comprehensive Cancer Center, University of North Carolina, Chapel Hill, North Carolina 27599, USA.
Joseph S
JS
Harrison
Research Institute of Molecular Pathology (IMP), Vienna Biocenter (VBC), 1030 Vienna, Austria.
Iain F
IF
Davidson
Research Institute of Molecular Pathology (IMP), Vienna Biocenter (VBC), 1030 Vienna, Austria.
Renping
R
Qiao
Department of Systems Biology, Harvard Medical School, Boston, MA, 02115, USA.
Ying
Y
Lu
Max Planck Institute for Biophysical Chemistry, 37077 Göttingen, Germany.
Prakash
P
Dube
Department of Structural Biology, St. Jude Children's Research Hospital, Memphis, TN 38105, USA.
Michael R
MR
Brunner
Department of Structural Biology, St. Jude Children's Research Hospital, Memphis, TN 38105, USA.
Christy R R
CRR
Grace
Department of Structural Biology, St. Jude Children's Research Hospital, Memphis, TN 38105, USA.
Darcie J
DJ
Miller
Max Planck Institute for Biophysical Chemistry, 37077 Göttingen, Germany.
David
D
Haselbach
Research Institute of Molecular Pathology (IMP), Vienna Biocenter (VBC), 1030 Vienna, Austria.
Marc A
MA
Jarvis
Department of Structural Biology, St. Jude Children's Research Hospital, Memphis, TN 38105, USA.
Masaya
M
Yamaguchi
Department of Structural Biology, St. Jude Children's Research Hospital, Memphis, TN 38105, USA.
David
D
Yanishevski
Research Institute of Molecular Pathology (IMP), Vienna Biocenter (VBC), 1030 Vienna, Austria.
Georg
G
Petzold
Donnelly Centre for Cellular and Biomolecular Research, University of Toronto, 160 College Street, Toronto, Ontario M5S3E1, Canada.
Sachdev S
SS
Sidhu
Department of Biochemistry and Biophysics and Lineberger Comprehensive Cancer Center, University of North Carolina, Chapel Hill, North Carolina 27599, USA.
Brian
B
Kuhlman
Department of Systems Biology, Harvard Medical School, Boston, MA, 02115, USA.
Marc W
MW
Kirschner
Department of Cell Biology, Harvard Medical School, Boston, MA 02115, USA.
J Wade
JW
Harper
Research Institute of Molecular Pathology (IMP), Vienna Biocenter (VBC), 1030 Vienna, Austria. Electronic address: jan-michael.peters@imp.ac.at.
Jan-Michael
JM
Peters
Max Planck Institute for Biophysical Chemistry, 37077 Göttingen, Germany. Electronic address: hstark1@gwdg.de.
Holger
H
Stark
Department of Structural Biology, St. Jude Children's Research Hospital, Memphis, TN 38105, USA; Howard Hughes Medical Institute, Memphis, TN 38105, USA. Electronic address: brenda.schulman@stjude.org.
Brenda A
BA
Schulman
D000595:Amino Acid Sequence; D064173:Anaphase-Promoting Complex-Cyclosome / Q000737:chemistry* / Q000378:metabolism*; D055162:Biocatalysis; D020285:Cryoelectron Microscopy; D006801:Humans; D008958:Models, Molecular; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry; D013329:Structure-Activity Relationship; D025801:Ubiquitin / Q000378:metabolism*; D044763:Ubiquitin-Conjugating Enzymes / Q000378:metabolism*; D054875:Ubiquitination
D029701:Saccharomyces cerevisiae Proteins; D025801:Ubiquitin; C105156:UBE2C protein, human; D044763:Ubiquitin-Conjugating Enzymes; D064173:Anaphase-Promoting Complex-Cyclosome