Mutations that alter an Arg-Gly-Asp (RGD) sequence in the adenovirus type 2 penton base protein abolish its cell-rounding activity and delay virus reproduction in flat cells.
The adenovirus penton base protein has a cell rounding activity and may lyse endosomes during virus entry into the cytoplasm. We found that penton base that was expressed in Escherichia coli also caused cell rounding and that cells adhered to polystyrene wells that were coated with the protein. Mutant analysis showed that both properties required an Arg-Gly-Asp (RGD) sequence at residues 340 to 342 of penton base. In flat adherent cells, virus mutants with amino acid substitutions in the RGD sequence were delayed in virus reproduction and in the onset of viral DNA synthesis. In nonadherent or poorly spread cells, the kinetics of mutant virus reproduction were similar to those of wild-type adenovirus type 2. Expression of the mutant phenotype exclusively in the flat cells that we tested supports a model in which penton base interacts with an RGD-directed cell adhesion molecule during adenovirus uptake or uncoating.
Bai
M
M
Biology Department, Brookhaven National Laboratory, Upton, New York 11973.
Harfe
B
B
Freimuth
P
P
D000260:Adenoviruses, Human / Q000235:genetics / Q000502:physiology*; D000595:Amino Acid Sequence; D001483:Base Sequence; D002213:Capsid / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D036022:Capsid Proteins; D002448:Cell Adhesion; D002460:Cell Line; D004261:DNA Replication; D004279:DNA, Viral / Q000096:biosynthesis; D005816:Genetic Complementation Test; D014644:Genetic Variation; D006367:HeLa Cells; D006801:Humans; D007700:Kinetics; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D009838:Oligodeoxyribonucleotides; D009842:Oligopeptides / Q000235:genetics / Q000378:metabolism*; D011485:Protein Binding; D015183:Restriction Mapping; D017386:Sequence Homology, Amino Acid; D014779:Virus Replication
D036022:Capsid Proteins; D004279:DNA, Viral; D009838:Oligodeoxyribonucleotides; D009842:Oligopeptides; C043322:penton protein, adenovirus; C047981:arginyl-glycyl-aspartic acid