Avian polyomavirus (APV) causes an acute fatal disease in a variety of avian species. DNA laddering indicating apoptosis was demonstrated in APV-infected chicken embryo (CE) cells. DNA laddering, however, was not observed in Vero cells infected with mammalian polyomavirus simian virus 40. Expression of APV agnoprotein 1a and agnoprotein 1b induced apoptosis in insect cells and CE cells. An APV full-length plasmid transfected in CE cells induced apoptosis, and infectious virus was produced. After transfection of CE cells with a plasmid containing a mutated initiation codon for agnoprotein 1a and agnoprotein 1b, however, a considerably lower number of apoptotic cells was observed, and no infectious progeny was produced.