Proximity Labeling Identifies BIRC6 as a Ubiquitin Ligase for KRAS4A
- Description
KRAS is a frequently mutated oncogene in human cancers and a critical driver of high-mortality cancers such as lung, colon, and pancreatic adenocarcinoma. Recent advances in drug design have led to the development of multiple strategies to inactivate KRAS. Therefore, this study performed proximity labeling to map the differential interactomes of the KRAS splice variants, KRAS4A and KRAS4B. Among the KRAS4A-specific interactors is BIRC6, a large member of the inhibitor of apoptosis proteins. The interaction between KRAS4A and BIRC6 was validated by co-immunoprecipitation. GFP-tagged RAS proteins were expressed along with 3×FLAG-BIRC6 in HEK293 cells and immunoprecipitated. To establish the preference for KRAS4A over KRAS4B in ubiquitination by BIRC6 in intact cells and to avoid non-specific bands caused by the exogenous expression of 8×His-Ub, 6×His-KRAS4A-12V or 6×His-KRAS4B-12V was expressed in HEK293 cells with or without GFP-BIRC6. This dataset includes raw proteomic data for the proximity labeling screen and the identification of ubiquitin modifications of 6×His-KRAS4A.
Access
- Restrictions
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Free to All
- Instructions
- Raw proteomic data are available on Mass Spectrometry Interactive Virtual Environment (MassIVE) repository.
- Grant Support
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Ramon Areces Foundation/Ramon Areces Foundation