The nucleolus is the most prominent structure within the eukaryotic cell nucleus and is the site where ribosomal RNAs (5.8S, 18S, and 28S) are transcribed, processed, and assembled with ribosomal proteins to form ribosomal subunits. A role in ribosome biogenesis alone, however, does not account for the specific nucleolar localizations of tumor suppressor proteins, cell cycle regulator factors, and viral proteins. Certain proteins have also been shown to accumulate in the nucleolus only under specific metabolic conditions or at specific cell cycle stages. The use of green fluorescent protein fusions has recently revealed that several proteins localize to distinct subnucleolar compartments via specific targeting pathways. Meanwhile, photobleaching analyses indicate that most nucleolar proteins studied are continually exchanging between the nucleoplasm and the nucleolus. Proteomic studies have also highlighted the dynamic nature of the nucleolar proteome. Both protein composition and the morphology of subnucleolar compartments can change in response to alterations in the levels of transcription and phosphorylation and during mitosis. This review focuses on nucleolar dynamics, studied at each of the single protein, collective proteome, and subnucleolar organization levels.