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Adenovirus core protein V interacts with p32--a protein which is associated with both the mitochondria and the
1School of Biomedical Sciences, University of St Andrews, UK.
Abstract:
Adenovirus protein V is associated with the DNA-containing virus core and functions as a bridge between the capsid and the core. A yeast two-hybrid analysis performed with a human cDNA library using protein V as 'bait' selected a cellular protein, p32 --described previously as associated with the splicing factor ASF/SF2. By expression and purification of p32 and preparation of an antibody we confirmed the binding of p32 to V by a variety of methods including immune precipitation. We demonstrated that p32 was primarily located in the cytoplasm in association with mitochondria but could also be detected in the nucleus as distinct granules and tubules. By examining infected cells using confocal microscopy and immunofluorescence we were able to follow the intracellular locations of protein V and p32 and it is postulated that p32 is part of a system which imports proteins to the nucleus and that adenovirus hijacks this process to deliver its genome to the nucleus.
Insights
Adenovirus protein V interacts with cellular protein p32, which may be involved in nuclear import. This interaction suggests adenovirus hijacks cellular machinery for genome delivery.
Area of Science:
- Virology
- Cell Biology
- Molecular Biology
Background:
- Adenovirus protein V is crucial for the virus core structure, linking the capsid and core.
- Protein p32 is a known cellular factor associated with the splicing factor ASF/SF2.
- Understanding viral protein interactions with host factors is key to deciphering viral replication strategies.
Purpose of the Study:
- To identify cellular proteins interacting with Adenovirus protein V.
- To characterize the interaction between Adenovirus protein V and its binding partners.
- To elucidate the role of these interactions in the adenovirus life cycle.
Main Methods:
- Yeast two-hybrid analysis using Adenovirus protein V as bait.
- Expression and purification of p32.
- Antibody generation and validation.
- Immune precipitation assays.
- Confocal microscopy and immunofluorescence in infected cells.
Main Results:
- Adenovirus protein V specifically binds to the cellular protein p32.
- Protein p32 is localized in the cytoplasm with mitochondria and in the nucleus as granules and tubules.
- Intracellular co-localization studies revealed the movement of protein V and p32 within infected cells.
Conclusions:
- Adenovirus protein V interacts with host protein p32.
- Protein p32 is potentially involved in a cellular protein import system.
- Adenovirus may exploit this host pathway for nuclear genome delivery, facilitating viral infection.