K33-Linked Polyubiquitination of Coronin 7 by Cul3-KLHL20 Ubiquitin E3 Ligase Regulates Protein Trafficking.
Ubiquitin chains are formed as structurally distinct polymers via different linkages, and several chain types including K33-linkage remain uncharacterized. Here, we describe a role for K33-polyubiquitination in protein trafficking. We show that the Cullin 3 (Cul3) substrate adaptor KLHL20 is localized to the trans-Golgi network (TGN) and is important for post-Golgi trafficking by promoting the biogenesis of TGN-derived transport carriers. The Cul3-KLHL20 ubiquitin E3 ligase catalyzes a nondegradable, K33-linked polyubiquitination on coronin 7 (Crn7), which facilitates Crn7 targeting to TGN through a ubiquitin-dependent interaction with Eps15. Blockage of K33-chain formation, Crn7 ubiquitination, or disruption of Crn7-Eps15 interaction impairs TGN-pool F-actin assembly, a process essential for generating transport carriers. Enforced targeting of Crn7 to TGN bypasses the requirement of K33-ubiquitination for TGN-pool F-actin assembly and post-Golgi trafficking. Our study reveals a role of KLHL20-mediated K33-ubiquitination of Crn7 in post-Golgi transport and identifies a cellular recognition mechanism for this ubiquitin chain type.
Institute of Biological Chemistry, Academia Sinica, Taipei 115, Taiwan; Institute of Biochemical Sciences, College of Life Science, National Taiwan University, Taipei 106, Taiwan.
Wei-Chien
WC
Yuan
Institute of Biological Chemistry, Academia Sinica, Taipei 115, Taiwan.
Yu-Ru
YR
Lee
Institute of Biological Chemistry, Academia Sinica, Taipei 115, Taiwan.
Shu-Yu
SY
Lin
Institute of Biological Chemistry, Academia Sinica, Taipei 115, Taiwan.
Li-Ying
LY
Chang
Institute of Biological Chemistry, Academia Sinica, Taipei 115, Taiwan.
Yen Pei
YP
Tan
Institute of Biological Chemistry, Academia Sinica, Taipei 115, Taiwan.
Chin-Chun
CC
Hung
Institute of Biochemistry and Molecular Biology, National Yang Ming University, Taipei 112, Taiwan.
Jean-Cheng
JC
Kuo
Institute of Biological Chemistry, Academia Sinica, Taipei 115, Taiwan.
Cheng-Hsin
CH
Liu
Institute of Biological Chemistry, Academia Sinica, Taipei 115, Taiwan.
Mei-Yao
MY
Lin
Department of Molecular Biology, Howard Hughes Medical Institute, UT Southwestern Medical Center at Dallas, Dallas, TX 75390, USA.
Ming
M
Xu
Department of Molecular Biology, Howard Hughes Medical Institute, UT Southwestern Medical Center at Dallas, Dallas, TX 75390, USA.
Zhijian J
ZJ
Chen
Institute of Biological Chemistry, Academia Sinica, Taipei 115, Taiwan; Institute of Biochemical Sciences, College of Life Science, National Taiwan University, Taipei 106, Taiwan. Electronic address: rhchen@gate.sinica.edu.tw.
Ruey-Hwa
RH
Chen
D000199:Actins / Q000235:genetics / Q000378:metabolism; D048868:Adaptor Proteins, Signal Transducing; D033942:Adaptor Proteins, Vesicular Transport / Q000235:genetics / Q000378:metabolism*; D000818:Animals; D019556:COS Cells; D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism*; D002460:Cell Line; D002522:Chlorocebus aethiops; D044842:Cullin Proteins / Q000235:genetics / Q000378:metabolism*; D006056:Golgi Apparatus / Q000378:metabolism; D057809:HEK293 Cells; D006367:HeLa Cells; D006801:Humans; D008239:Lysine / Q000378:metabolism; D008840:Microfilament Proteins / Q000235:genetics / Q000378:metabolism*; D021381:Protein Transport; D044767:Ubiquitin-Protein Ligases / Q000235:genetics / Q000378:metabolism*; D054875:Ubiquitination; D021601:trans-Golgi Network / Q000378:metabolism
D000199:Actins; D048868:Adaptor Proteins, Signal Transducing; D033942:Adaptor Proteins, Vesicular Transport; D002352:Carrier Proteins; D044842:Cullin Proteins; C550011:KLHL20 protein, human; D008840:Microfilament Proteins; C072061:coronin proteins; D044767:Ubiquitin-Protein Ligases; D008239:Lysine