Adenovirus E1b region-coded 19K tumor antigen plays an important role in cell transformation in cooperation with the E1a proteins. We propose to investigate the role of 19K in adenovirus-mediated transformation using genetic and biochemical approaches. It appears that E1a-mediated transformation may be negatively regulated by other early viral genes. We will investigate the effect of the 19K gene relieving the transformation inhibitory effect of other early genes by DNA transfection and by using viral double mutants. Physical association of 19K with other early proteins will be studied by coimmunoprecipitation analysis of proteins cross-linked in living cells. Other early genes that interact with 19K protein will also be investigated using revertants of a 19K mutant, cyt6. The site of reversion will be mapped by marker rescue and by DNA sequence analysis. One of our major goals would be to identify cellular factors that physically interact with 19K during transformation. These proteins will be identified by coimmunoprecipitation analysis of proteins cross-linked in living cells over-express the 19K protein. The identity of the cellular proteins will be determinied by partial sequence determination of the genes coding for these proteins. To probe the structure and function of 19K, mutants with one or two amino acid substitutions or deletions will be constructed in the regions which appear to be functionally important. We have noted that a region of the 19K protein is homologous to a region of polyoma middle T (mT) antigen which is important for trans- formation. We will study the functional similarity between this 19K protein region and the corresponding mT protein region by domain substitution. We have observed that 19K plays a role in calcium mobilization. We will determine whether 19K plays a role in signal transduction at the level of generation of second messengers by studying the turnover of phosphoinositotides. As 19K appears to alter the architecture of the endoplasmic reticulum, we will also investigate whether 19K plays a role in calcium release from the endoplasmic reticulum by using isolated ER membrane vesicles in the presence of inositol tris- phosohate (InsP3). The 19K protein contains an amphipathic acidic domain similar to the calcium regulating domain of epidermal growth factor receptor (EGFR) and related receptors. To determine whether the acidic domain of 19K is functionally similar to this EGFR domain, we will construct a chimeric EGFR in which the acidic domain is replaced with the corresponding domain of 19K. The role of the chimeric receptor in calcium mobilization and ligand-dependent internalization will be determined. The proposed studies should facilitate understanding of the biochemical mechanism of 19K-mediated transformation.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
5R01CA033616-14
Application #
3171427
Study Section
Experimental Virology Study Section (EVR)
Project Start
1979-07-01
Project End
1995-11-30
Budget Start
1992-12-01
Budget End
1993-11-30
Support Year
14
Fiscal Year
1993
Total Cost
Indirect Cost
Name
Saint Louis University
Department
Type
Schools of Medicine
DUNS #
City
Saint Louis
State
MO
Country
United States
Zip Code
63103
Vijayalingam, S; Subramanian, T; Zhao, Ling-Jun et al. (2016) The Cellular Protein Complex Associated with a Transforming Region of E1A Contains c-MYC. J Virol 90:1070-9
Subramanian, T; Vijayalingam, S; Kuppuswamy, M et al. (2015) Interaction of cellular proteins with BCL-xL targeted to cytoplasmic inclusion bodies in adenovirus infected cells. Virology 483:21-31
Zhao, Ling-Jun; Subramanian, T; Vijayalingam, S et al. (2014) CtBP2 proteome: Role of CtBP in E2F7-mediated repression and cell proliferation. Genes Cancer 5:31-40
Vijayalingam, S; Kuppusamy, Mohan; Subramanian, T et al. (2014) Evaluation of apoptogenic adenovirus type 5 oncolytic vectors in a Syrian hamster head and neck cancer model. Cancer Gene Ther 21:228-237
Subramanian, T; Zhao, Ling-Jun; Chinnadurai, G (2013) Interaction of CtBP with adenovirus E1A suppresses immortalization of primary epithelial cells and enhances virus replication during productive infection. Virology 443:313-20
Kuppuswamy, Mohan; Subramanian, T; Kostas-Polston, Elizabeth et al. (2013) Functional similarity between E6 proteins of cutaneous human papillomaviruses and the adenovirus E1A tumor-restraining module. J Virol 87:7781-6
Vijayalingam, S; Chinnadurai, G (2013) Adenovirus L-E1A activates transcription through mediator complex-dependent recruitment of the super elongation complex. J Virol 87:3425-34
Chinnadurai, G (2011) Opposing oncogenic activities of small DNA tumor virus transforming proteins. Trends Microbiol 19:174-83
Vijayalingam, S; Pillai, Sreeraj G; Rashmi, Ramachandran et al. (2010) Overexpression of BH3-Only Protein BNIP3 Leads to Enhanced Tumor Growth. Genes Cancer 1:964-71
Komorek, Jessica; Kuppuswamy, Mohan; Subramanian, T et al. (2010) Adenovirus type 5 E1A and E6 proteins of low-risk cutaneous beta-human papillomaviruses suppress cell transformation through interaction with FOXK1/K2 transcription factors. J Virol 84:2719-31

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