Fused in sarcoma (FUS) is an RNA/DNA binding protein that normally resides in the nucleus. However, FUS forms pathologic cytoplasmic inclusions in two neurodegenerative disorders, frontotemporal lobar degeneration (FTLD) and amyotrophic lateral sclerosis (ALS). While a majority of ALS cases with FUS pathology can be explained by pathogenic mutations in the FUS gene, the vast majority of FTLD-FUS cases are not caused by FUS mutations and the reason why FUS forms inclusions is unknown. Therefore, identification of other non-genetic mechanisms that cause FUS to accumulate in the cytoplasm is crucial to understanding FTLD and ALS pathogenesis. To this end, DNA damage is known to trigger DNA-PK to phosphorylate FUS at N-terminal residues leading to FUS accumulation in the cytoplasm. However, the functional consequences of FUS phosphorylation are unknown. In this study, we performed proximity-dependent biotin labeling via ascorbate peroxidase 2 (APEX2) paired with mass spectrometry to investigate whether phosphorylation shifts the FUS interactome and protein function. Data are available via ProteomeXchange with identifier PXD026578. We identified a highly interrelated interactome between wild-type, phosphomimetic FUS (a proxy for phosphorylated FUS), and the pathogenic ALS-linked mutant P525L FUS. We demonstrate that expression of phosphomimetic FUS shifts the FUS interactome toward more cytoplasmic functions including mediation of mRNA metabolism and translation. Our findings reveal that phosphorylation of FUS may disrupt homeostatic translation and mRNA metabolism. These results highlight the importance of phosphorylation as a modulator of FUS interactions and functions with a potential link to disease pathogenesis.