Details

Autor(en) / Beteiligte

• Oppenheim, Tal
• Radzinski, Meytal
• Braitbard, Merav
• Brielle, Esther S.
• Yogev, Ohad
• Goldberger, Eliya
• Yesharim, Yarden
• Ravid, Tommer
• Schneidman-Duhovny, Dina
• Reichmann, Dana

Titel

The Cdc48 N-terminal domain has a molecular switch that mediates the Npl4-Ufd1-Cdc48 complex formation

Ist Teil von

• Structure (London), 2023-07, Vol.31 (7), p.764-779.e8

Ort / Verlag

United States: Elsevier Ltd

Erscheinungsjahr

2023

Quelle

MEDLINE

Beschreibungen/Notizen

• 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. [Display omitted] •XL-MS and integrative modeling reveals the architecture of the Npl4 and Ufd1 complex•Binding of the N-terminal domain of Cdc48 alters dynamics of Npl4-Ufd1•Cysteine 115 mediates binding between Cdc48-NTD and Npl4-Ufd1•Mutation of Cys115 to Serine alters Cdc48 activity in yeast cells The Cdc48-Npl4-Ufd1 complex is central for protein quality control in cells. Oppenheim, et al. provide insights into the architecture and dynamics of the ternary complex, using crosslinking mass spectrometry and integrative structural modeling. They reveal the importance of a binding interface cysteine residue on the structure and activity of the Cdc48-Npl4-Ufd1 complex.