Formation in vitro of the pTP-dCMP initiation complex of human adenovirus type 12.
We report the covalent addition of [32P]dCMP to a protein from group A adenovirus 12 (Ad12)-infected human (KB) cells in vitro, using crude extracts. Synthesis of the 60K protein-dCMP complex required a DNA template containing a terminally located adenovirus replication origin; the protein-dCMP bond was alkali-labile but acid-stable. We therefore conclude that this product is the Ad12 terminal protein precursor (pTP)-dCMP initiation complex for DNA replication. Synthesis of Ad12 pTP-dCMP was specific for dCTP but was stimulated by dATP. In contrast to Ad2, the Ad12 initiation reaction required ATP. Antipeptide antiserum targeted to Ad DNA polymerase inhibited Ad12 pTP-dCMP synthesis in vitro, providing evidence that Ad DNA polymerase catalyse dCMP addition to pTP during initiation.
Chowrira
B M
BM
Department of Biological Sciences, Illinois State University, Normal 61761.
Zhao
J S
JS
Lucher
L A
LA
D000260:Adenoviruses, Human / Q000378:metabolism*; D000595:Amino Acid Sequence; D003570:Cytidine Triphosphate / Q000378:metabolism; D003843:Deoxycytidine Monophosphate / Q000378:metabolism*; D006801:Humans; D015151:Immunoblotting; D007624:KB Cells; D008969:Molecular Sequence Data; D014764:Viral Proteins / Q000378:metabolism*; D014779:Virus Replication
D014764:Viral Proteins; D003843:Deoxycytidine Monophosphate; D003570:Cytidine Triphosphate