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Quantitative analysis of regions of adenovirus E1A products involved in interactions with cellular proteins
D Barbeau1, R C Marcellus, S Bacchetti
1Department of Pathology, McMaster University, Hamilton, Ont., Canada.
Abstract:
Human adenovirus E1A proteins and oncogene products of several other DNA tumour viruses derive much of their oncogenic potential from interactions with cellular polypeptides. E1A proteins form complexes with p105Rb and a related p107 polypeptide, and with at least three other proteins (p60cycA, p130, and p300); all may be required for cell transformation. Using a series of E1A deletion mutants, we have carried out a quantitative analysis of the binding patterns of cellular proteins to E1A products. Binding of most of the proteins was affected at least partially by mutations within the amino terminal 25 residues, amino acids 36-69 within conserved region 1 (CR1), and residues 121-138 in conserved region 2 (CR2). However, the specific binding characteristics of each protein varied considerably. p300 was the only species for which binding was totally eliminated by deletions at the amino terminus. Removal of regions within CR1 eliminated binding of all species except p107 and p60cycA. Deletion of portions of CR2 reduced or eliminated binding of all proteins except p300. Thus, whereas cellular polypeptides generally were found to interact with the same three regions of E1A proteins, specific interactions varied considerably.
Insights
Human adenovirus E1A proteins interact with cellular proteins for oncogenic potential. Specific binding regions on E1A proteins were identified using deletion mutants, revealing varied interaction patterns.
Area of Science:
- Molecular Biology
- Virology
- Oncology
Background:
- Oncogenic potential of DNA tumor viruses often stems from interactions between viral oncoproteins and cellular proteins.
- Adenovirus E1A proteins are known to form complexes with cellular proteins like p105Rb, p107, p60cycA, p130, and p300, which are crucial for cell transformation.
Purpose of the Study:
- To quantitatively analyze the binding patterns of cellular proteins to human adenovirus E1A proteins.
- To identify specific regions within E1A proteins responsible for binding various cellular polypeptides.
Main Methods:
- Utilized a series of E1A deletion mutants to systematically assess protein-protein interactions.
- Performed quantitative analysis of cellular protein binding to different E1A mutant constructs.
Main Results:
- Binding of most cellular proteins to E1A was affected by mutations in the N-terminus, conserved region 1 (CR1), and conserved region 2 (CR2).
- Specific binding profiles varied: p300 binding was exclusively sensitive to N-terminal deletions, while CR1 and CR2 deletions differentially impacted other protein bindings.
- CR1 deletions abolished binding for most proteins except p107 and p60cycA; CR2 deletions affected all proteins except p300.
Conclusions:
- Cellular polypeptides generally interact with three key regions of E1A proteins.
- The specific interactions between E1A proteins and cellular polypeptides are highly nuanced and region-specific, contributing to the complexity of viral oncogenesis.
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