Polyomaviruses (Pys) are nonenveloped DNA tumor viruses that include the murine polyomavirus (mPy), simian virus 40 (SV40), and the human BK, JC, KI, WU, and Merkel Cell viruses. To cause infection, Pys must enter host cells and navigate through various intracellular compartments, where they undergo sequential conformational changes enabling them to uncoat and deliver the DNA genome into the nucleus. The ensuing transcription and replication of the genome leads to lytic infection or cell transformation. In recent years, a more coherent understanding of how Pys are transported from the plasma membrane to the nucleus is starting to emerge. This review will focus on the decisive steps of Py entry, including engagement of the host cell receptor, targeting to the endoplasmic reticulum (ER), penetration across the ER membrane, nuclear entry, and genome release. Strikingly, a number of these steps resemble the intoxication pathway of the AB(5) bacterial toxins. Thus, as Pys and bacterial toxins hijack similar cellular machineries during infection, a general principle appears to guide their entry into host cells.