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Cellular components interact with adenovirus type 5 minimal DNA packaging domains
1Department of Molecular Genetics and Microbiology, Health Sciences Center, State University of New York at Stony Brook, Stony Brook, New York 11794, USA.
Journal of Virology
|July 11, 1998
Summary
Adenovirus type 5 DNA packaging relies on specific elements (A repeats) that bind cellular factors. A newly identified P-complex binds these elements and the viral genome
Area of Science:
- Virology
- Molecular Biology
- Genetics
Background:
- Adenovirus type 5 DNA packaging initiates at the genome's left end, requiring a cis-acting domain with multiple redundant packaging elements (A repeats I-VII).
- Functionally important A repeats (AI, AII, AV, AVI) possess bipartite consensus motifs with AT-rich and CG-rich cores.
- Previous studies identified a minimal packaging domain (AV, AVI, AVII) supporting functional independence of cis-acting sequences.
Purpose of the Study:
- To characterize multimeric individual packaging elements as minimal domains conferring viability and packaging activity to viruses with truncated left ends.
- To investigate the functional roles and relative importance of different packaging elements.
- To identify and characterize cellular factors that interact with these packaging elements.
Main Methods:
- Construction and analysis of mutant viruses with multimeric individual packaging elements and gross truncations.
- In vitro characterization of cellular binding activities interacting with minimal packaging domains.
- Assessment of binding affinity and correlation with in vivo packaging activity.
Main Results:
- Multimeric packaging elements function as minimal domains, enabling packaging in viruses with truncated left ends, allowing direct assessment of element roles.
- Two cellular binding activities, P-complex and chicken ovalbumin upstream promoter transcription factor, interact with minimal packaging domains.
- P-complex binding affinity correlates with in vivo packaging ability and it also binds the viral genome's replication origin.
Conclusions:
- Cellular factors, including P-complex, are integral components of the adenovirus packaging machinery.
- The dual specificity of P-complex for packaging elements and replication sequences suggests a role for left-end replication sequences in DNA encapsidation.
- These findings highlight the involvement of cellular factors in viral DNA packaging and replication coordination.