Unleashed condensation by recurrent mutations of an epigenetic regulator promotes cancer

ABSTRACT: The pathological mechanisms and significance for the prevalent mutation hotspots in disordered protein regions are poorly understood. ASXL1 is an obligate co-factor for BAP1 in H2AK119 deubiquitination. ASXL1 mutations are very frequent in myeloid malignancies, and are mostly C-terminal truncating mutations concentrated in a specific disordered region of ASXL1. ASXL1 truncations are gain-of-function mutations that promote myeloid malignancies, but the underlying mechanisms remain poorly understood. Here we show that the frequently truncated mutants of ASXL1 possess an intrinsic property of forming phase-separated biomolecular condensates, and this property is normally suppressed by the frequently deleted regions. A disease-mutant of the endogenous ASXL1 in leukemia cells forms dynamic nuclear co-condensates with other endogenous factors important for gene activation. While the ASXL1 disease-mutant can greatly enhance H2A deubiquitination activity of BAP1 in cells and in a purified system, enhance the growth of myeloid leukemia cell growth, and promote hematopoietic colony formation and leukemogenesis in a mouse transplant model by turning on myeloid leukemogenic transcriptional programs, substitution of residues important for condensation disrupted or impaired these abilities, suggesting that the condensation property is crucially important for the ASXL1 mutants in promoting cancer. Moreover, we discover that the conserved negative charges in the highly disordered and frequently deleted region on ASXL1 suppress the condensation of the wild type ASXL1. Charge-neutralizing mutations in this region restores condensation of the full-length ASXL1, and renders the full-length protein capable of promoting leukemogenesis-associated activity and gene expression. Biochemical, biophysical, and simulation analyses suggest the intramolecular interactions normally mask the N-terminal region in engaging intermolecular interactions required for phase separation, and disease truncations escape from the regulatory interactions and unleash the phase separation property to form nuclear hubs to promote expression of tumorigenic gene programs. Finally, by showing a striking correlation of the mutation frequencies with the condensation properties and leukemogenesis activity for a series of human patient mutations, we suggest that dysregulation of condensation is a likely a central mechanism for ASXL1 mutations in promoting myeloid malignancies. This may also be an overlooked mechanism for the prevalent hotspot disease mutations in the disordered proteomes.

ORGANISM(S): Mus musculus

PROVIDER: GSE264442 | GEO | 2026/04/23

REPOSITORIES: GEO