TY - JOUR
T1 - The Cdc48 N-terminal domain has a molecular switch that mediates the Npl4-Ufd1-Cdc48 complex formation
AU - Oppenheim, Tal
AU - Radzinski, Meytal
AU - Braitbard, Merav
AU - Brielle, Esther S.
AU - Yogev, Ohad
AU - Goldberger, Eliya
AU - Yesharim, Yarden
AU - Ravid, Tommer
AU - Schneidman-Duhovny, Dina
AU - Reichmann, Dana
N1 - Publisher Copyright:
© 2023 Elsevier Ltd
PY - 2023/7/6
Y1 - 2023/7/6
N2 - Cdc48 (VCP/p97) is a major AAA-ATPase involved in protein quality control, along with its canonical cofactors Ufd1 and Npl4 (UN). Here, we present novel structural insights into the interactions within the Cdc48-Npl4-Ufd1 ternary complex. Using integrative modeling, we combine subunit structures with crosslinking mass spectrometry (XL-MS) to map the interaction between Npl4 and Ufd1, alone and in complex with Cdc48. We describe the stabilization of the UN assembly upon binding with the N-terminal-domain (NTD) of Cdc48 and identify a highly conserved cysteine, C115, at the Cdc48-Npl4-binding interface which is central to the stability of the Cdc48-Npl4-Ufd1 complex. Mutation of Cys115 to serine disrupts the interaction between Cdc48-NTD and Npl4-Ufd1 and leads to a moderate decrease in cellular growth and protein quality control in yeast. Our results provide structural insight into the architecture of the Cdc48-Npl4-Ufd1 complex as well as its in vivo implications.
AB - Cdc48 (VCP/p97) is a major AAA-ATPase involved in protein quality control, along with its canonical cofactors Ufd1 and Npl4 (UN). Here, we present novel structural insights into the interactions within the Cdc48-Npl4-Ufd1 ternary complex. Using integrative modeling, we combine subunit structures with crosslinking mass spectrometry (XL-MS) to map the interaction between Npl4 and Ufd1, alone and in complex with Cdc48. We describe the stabilization of the UN assembly upon binding with the N-terminal-domain (NTD) of Cdc48 and identify a highly conserved cysteine, C115, at the Cdc48-Npl4-binding interface which is central to the stability of the Cdc48-Npl4-Ufd1 complex. Mutation of Cys115 to serine disrupts the interaction between Cdc48-NTD and Npl4-Ufd1 and leads to a moderate decrease in cellular growth and protein quality control in yeast. Our results provide structural insight into the architecture of the Cdc48-Npl4-Ufd1 complex as well as its in vivo implications.
KW - CDC48
KW - CL-MS
KW - NPL4
KW - UFD1
KW - crosslinking coupled with MS (XL-MS)
KW - integrative modeling
KW - protein quality control
KW - structural mass spectrometry
UR - http://www.scopus.com/inward/record.url?scp=85163932903&partnerID=8YFLogxK
U2 - 10.1016/j.str.2023.05.014
DO - 10.1016/j.str.2023.05.014
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C2 - 37311459
AN - SCOPUS:85163932903
SN - 0969-2126
VL - 31
SP - 764-779.e8
JO - Structure
JF - Structure
IS - 7
ER -