The inositol 5-phosphatase SHIP1 is a negative regulator of signaling processes in haematopoietic cells. By converting PI(3,4,5)P3 to PtdIns(3,4)P2 at the plasma membrane, SHIP1 modifies PI3-kinase mediated signaling. We have recently demonstrated that SHIP1 is a nucleo-cytoplasmic shuttling protein and SHIP1 nuclear puncta partially colocalize with FLASH, a component of nuclear bodies. In this study, we demonstrate that endogenous SHIP1 localizes to intranucleolar regions of both normal and leukemic haematopoietic cells. In addition, we report that ectopically expressed SHIP1 accumulates in nucleolar cavities and colocalizes with the tumor suppressor protein p53 and components of PML nuclear bodies (e.g. SP100, SUMO-1 and CK2). Moreover, SHIP1 also colocalizes in nucleolar cavities with components of the ubiquitin-proteasome pathway. By using confocal microscopy data, we generated 3D-models revealing the enormous extent of the SHIP1 aggresomes in the nucleolus. Furthermore, treatment of cells with the proteasome inhibitor MG132 causes an enlargement of nucleolar SHIP1 containing structures. Unexpectedly, this accumulation can be partially prevented by treatment with the inhibitor of nuclear protein export Leptomycin B. In recent years, several proteins aggregating in nucleolar cavities were shown to be key factors of neurodegenerative diseases and cancerogenesis. Our findings support current relevance of nuclear localized SHIP1.
Keywords: DFC, dense fibrillar component; DIC, Differential interference contrast; EGFP, enhanced green fluorescent protein; FC, fibrillar center; GC, granular component; LMB, leptomycin B; MG132; NES, nuclear export signal; PBMC, Peripheral Blood Mononuclear Cell; PML bodies; PML, Promyelocytic Leukemia; PtdIns(3, 4)P2, phosphatidylinositol-(3, 4)-bisphosphate; PtdIns(3, 4, 5)P3, phosphatidylinositol-(3, 4, 5)-trisphosphate; RNA pol, RNA polymerase; SHIP1; SHIP1, src homology 2 domain-containing inositol phosphatase 1; UPP, ubiquitin-proteasome pathway.; aggresome; cancer; leptomycin B; nucleolar cavities; nucleus; p53; ubiquitin proteasome pathway.