Nuclear bodies (NBs) are membraneless organelles that play important roles in genome functioning. A specific type of NBs known as interphase prenucleolar bodies (iPNBs) are formed in the nucleoplasm after hypotonic stress from partially disassembled nucleoli. iPNBs are then disassembled, and the nucleoli are reformed simultaneously. Here, we show that diffusion of B23 molecules from iPNBs but not fusion of iPNBs with the nucleoli contributes to the transfer of B23 from iPNBs to the nucleoli. Maturation of pre-rRNAs and the subsequent outflow of mature rRNAs from iPNBs lead to the disassembly of iPNBs. We found that B23 transfer was dependent on the synthesis of pre-rRNA molecules in nucleoli; these pre-rRNA molecules interacted with B23 and led to its accumulation within nucleoli. The transfer of B23 between iPNBs and nucleoli was accomplished via a nucleoplasmic pool of B23, and increased nucleoplasmic B23 content retarded disassembly, while B23 depletion accelerated disassembly. Our results suggest that iPNB disassembly and nucleolus assembly may be coupled via RNA-dependent exchange of nucleolar proteins, creating a highly dynamic system with long-distance correlations between spatially distinct processes.