cis and trans requirements for the selective packaging of adenovirus type 5 DNA.
Polar packaging of adenovirus DNA into virions is dependent on the presence of cis-acting sequences at the left end of the viral genome. Our previous analyses demonstrated that the adenovirus type 5 (Ad5) packaging domain (nucleotides 194 to 358) is composed of at least five elements that are functionally redundant. A repeated sequence, termed the A repeat, was associated with packaging function. Here we report a more detailed analysis of the requirements for the selective packaging of Ad5 DNA. By introducing site-directed point mutations into specific A repeat sequences, we demonstrate that the A repeats represent cis-acting functional components of the packaging signal. Additional elements, located outside the originally defined packaging domain boundaries and that resemble the A repeat consensus sequence, also are capable of promoting the packaging of viral DNA. The cis-acting components of the packaging signal appear to be subject to certain spatial constraints for function, possibly reflecting a necessity for the coordinate binding of packaging proteins to these sites. In agreement with this idea, we present evidence that the interaction of a limiting trans-acting factor(s) with the packaging domain in vivo is required for efficient encapsidation of the Ad5 genome.
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Department of Microbiology, State University of New York, Stony Brook 11794-7621.
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D000260:Adenoviruses, Human / Q000235:genetics* / Q000502:physiology; D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D002467:Cell Nucleus / Q000502:physiology; D004247:DNA / Q000235:genetics / Q000302:isolation & purification; D004279:DNA, Viral / Q000235:genetics* / Q000302:isolation & purification; D016679:Genome, Viral; D006801:Humans; D008969:Molecular Sequence Data; D016254:Mutagenesis, Insertional; D016297:Mutagenesis, Site-Directed; D009690:Nucleic Acid Conformation; D012091:Repetitive Sequences, Nucleic Acid; D014162:Transfection; D014771:Virion / Q000235:genetics / Q000502:physiology; D014779:Virus Replication
D004279:DNA, Viral; D004247:DNA