Molecular Basis for CPC-Sgo1 Interaction: Implications for Centromere Localisation and Function of the CPCMolecular Basis for CPC-Sgo1 Interaction: Implications for Centromere Localisation and Function of the CPC

ABSTRACT: Sgo1 makes multipartite interactions with CPC subunits Previous studies have suggested that the Sgo1-CPC (Chromosomal Passenger Complex) interaction is mediated via the N-terminal coiled-coil of Sgo1 and Borealin. However, our structural data revealed that the very N-terminus of Sgo1 can interact with the BIR domain of Survivin. Together, these studies suggest that multi-partite interactions between Sgo1 and different CPC subunits could facilitate CPC-Sgo1 complex formation. To gain further structural insights, we performed chemical cross-linking of the CPCISB10-280-Sgo11-415 complex using a zero-length cross-linker, 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC), followed by mass spectrometry analysis. Cross-linking-mass spectrometry (CLMS) data showed that: (1) consistent with our previous observations, the N-terminal region of Sgo1 (amino acids 1-34) makes extensive contacts with Survivin BIR domain (amino acids 18-89); (2) the N-terminal coiled-coil of Sgo1 (amino acids 10-120) interacts with the CPC triple helical bundle; (3) consistent with previous findings, the N-terminal coiled-coil also contacts the Borealin dimerisation domain; and (4) the Sgo1 region beyond the N-terminal coil-coil region, which is predicted to be unstructured, contacts both Survivin and Borealin with most contacts confined to the Sgo1 central region spanning amino acids (aa) 180-300. Thus, our cross-linking results suggests that Sgo1 interacts with CPC, mainly via two regions, the N-terminal coiled-coil domain and the unstructured central region.

INSTRUMENT(S): Orbitrap Fusion Lumos

ORGANISM(S): Homo Sapiens (human)

SUBMITTER: Juan Zou  

LAB HEAD: Juri Rappsilber

PROVIDER: PXD028433 | Pride | 2022-06-29

REPOSITORIES: Pride