Analysis of the interaction of the adenovirus L1 52/55-kilodalton and IVa2 proteins with the packaging sequence in vivo and in vitro.
We previously showed that the adenovirus IVa2 and L1 52/55-kDa proteins interact in infected cells and the IVa2 protein is part of two virus-specific complexes (x and y) formed in vitro with repeated elements of the packaging sequence called the A1-A2 repeats. Here we demonstrate that both the IVa2 and L1 52/55-kDa proteins bind in vivo to the packaging sequence and that each protein-DNA interaction is independent of the other. There is a strong and direct interaction of the IVa2 protein with DNA in vitro. This interaction is observed when probes containing the A1-A2 or A4-A5 repeats are used, but it is not found by using an A5-A6 probe. Furthermore, we show that complex x is likely a heterodimer of IVa2 and an unknown viral protein, while complex y is a monomer or multimer of IVa2. No in vitro interaction of purified L1 52/55-kDa protein with the packaging sequence was found, suggesting that the L1 52/55-kDa protein-DNA interaction may be mediated by an intermediate protein. Results support roles for both the L1 52/55-kDa and IVa2 proteins in DNA encapsidation.
Perez-Romero
Pilar
P
Department of Microbiology and Immunology and Comprehensive Cancer Center, University of Michigan Medical School, 1500 E. Medical Center Dr., Ann Arbor, MI 48109-0942, USA.
Tyler
Ryan E
RE
Abend
Johanna R
JR
Dus
Monica
M
Imperiale
Michael J
MJ
D000260:Adenoviruses, Human / Q000235:genetics / Q000502:physiology*; D002460:Cell Line; D042003:DNA Packaging / Q000502:physiology*; D004279:DNA, Viral / Q000096:biosynthesis / Q000235:genetics; D006801:Humans; D008970:Molecular Weight; D014764:Viral Proteins / Q000096:biosynthesis / Q000302:isolation & purification / Q000378:metabolism*; D019065:Virus Assembly
D004279:DNA, Viral; D014764:Viral Proteins; C056232:iva2 protein, adenovirus