Serial Capture Affinity Purification and Integrated Structural Modeling of the H3K4me3 Binding and DNA Damage Related...

Area of Science:

• Molecular Biology
• Epigenetics
• Proteomics

Background:

• WD repeat domain 76 (WDR76) is a multifunctional protein with a complex interaction network.
• Dissecting specific WDR76 protein complexes is crucial for understanding its cellular functions.
• The Serial Capture Affinity Purification (SCAP) method allows for the isolation of specific protein complexes using dual baits.

Approach:

• Applied the SCAP method to isolate and analyze the WDR76:SPIN1 complex.
• Utilized crosslinking mass spectrometry to determine the structural model of the complex.
• Conducted interaction network analysis of copurifying proteins.

Key Points:

• The WDR76:SPIN1 complex was successfully isolated and characterized.
• SPIN1 recognizes the H3K4me3 epigenetic mark while interacting with WDR76.
• The WDR76:SPIN1 complex interacts with core histones.

Conclusions:

• An integrated structural model revealed SPIN1's dual recognition of H3K4me3 and WDR76.
• The WDR76:SPIN1 complex is potentially involved in the DNA damage response.
• This study elucidates a specific subpopulation of WDR76 and its functional implications.