Phosphatidylinositol 4,5 bisphosphate (PIP2) is the most abundant eukaryotic nuclear phosphoinositide. PIP2 is found in the nuclear speckles (NS), nucleoli (NL), and nuclear lipid islets (NLIs) and participates in the RNA polymerase II (Pol2) transcription. The formation of phase-separated nuclear sub-compartments, such as NS and NL, as well as PIP2 localization, depends on RNA. Thus, we studied if the PIP2 and RNA cooperate in the establishment of nuclear architecture. We identified the RNA-dependent PIP2-associated (RDPA) nuclear proteome in human cells. RDPA proteome participates in all steps of gene expression, and typical RDPA protein possesses RNA binding and phase separation capacity. RDPA proteins associate with PIP2 by electrostatic interactions, which depend on intact RNA and the phase separation capacity of the participating proteins. Moreover, we identified that intrinsically disordered regions (IDRs) with polybasic PIP2-binding K/R-motifs are common structural features of RDPA proteins and confirmed that manipulation of the PIP2 level changes the nuclear localization of RDPA proteins. We showed that PIP2 spatiotemporally orchestrates nuclear processes through association with RNA and RDPA proteins and their capacity to phase separate, which is the first evidence that PIP2 is crucial for establishment of functional nuclear architecture competent for gene expression.