Journal Article

Proteogenomic analysis reveals cytoplasmic sequestration of RUNX1 by the acute myeloid leukemia–initiating CBFB::MYH11 oncofusion protein

Erdmann-Gilmore, Petra, Day, Ryan B., Miller, Christopher A., Katerndahl, Casey D.S., Sprung, Robert W., Ramakrishnan, Sai Mukund, Ley, Timothy J., Hickman, Julia A., Townsend, R. Reid, Xu, Ziheng, Ferraro, Francesca, Mi, Yiling

2024

Overview

Several canonical translocations produce oncofusion genes that can initiate acute myeloid leukemia (AML). Although each translocation is associated with unique features, the mechanisms responsible remain unclear. While proteins interacting with each oncofusion are known to be relevant for how they act, these interactions have not yet been systematically defined. To address this issue in an unbiased fashion, we fused a promiscuous biotin ligase (TurboID) in-frame with 3 favorable-risk AML oncofusion cDNAs (PML::RARA, RUNX1::RUNX1T1, and CBFB::MYH11) and identified their interacting proteins in primary murine hematopoietic cells. The PML::RARA- and RUNX1::RUNX1T1-TurboID fusion proteins labeled common and unique nuclear repressor complexes, implying their nuclear localization. However, CBFB::MYH11-TurboID-interacting proteins were largely cytoplasmic, probably because of an interaction of the MYH11 domain with several cytoplasmic myosin-related proteins. Using a variety of methods, we showed that the CBFB domain of CBFB::MYH11 sequesters RUNX1 in cytoplasmic aggregates; these findings were confirmed in primary human AML cells. Paradoxically, CBFB::MYH11 expression was associated with increased RUNX1/2 expression, suggesting the presence of a sensor for reduced functional RUNX1 protein, and a feedback loop that may attempt to compensate by increasing RUNX1/2 transcription. These findings may have broad implications for AML pathogenesis.

• Publisher: American Society for Clinical Investigation
• Subject: Acute myeloid leukemia / Animals / Antibodies / B cells / Binding sites / Biotin / Core Binding Factor Alpha 2 Subunit - genetics / Core Binding Factor beta Subunit / DNA methylation / Genes / Genetic aspects / Hematopoietic stem cells / Humans / Labeling / Leukemia / Leukemia, Myeloid, Acute - pathology / Ligases / Localization / Mass spectrometry / Mice / Muscle proteins / Myosin / Myosin Heavy Chains - genetics / Oncogene Proteins, Fusion - genetics / Oncogene Proteins, Fusion - metabolism / Oncology / Pathogenesis / Proteins / Proteogenomics / Runx1 protein / Scientific imaging / Transcription factors / Translocation, Genetic