Abstract
C-terminal binding protein (CtBP) associates with adenovirus early region 1A (AdE1A) proteins through a highly conserved PXDLS motif located very close to its C-terminus in conserved region 4. To try to understand the importance of this interaction for the virus a point mutation in the CtBP binding site of Adl2ElA (P→S at amino acid 255) was engineered. The mutant Ad12E1A DNA (Ad12E1A6f) encoded a protein temperature sensitive (ts) for transformation of baby rat kidney cells when in combination with Ad12ElB. At 33°C transformation frequency was comparable to wt. At 37° and 38.5° transformants appeared as larger epithelioid cells and colonies senesced relatively rapidly. When the Ad12 6f AdE1A was incorporated into a mutant virus it caused a marked reduction in its ability to replicate with only Ad12E1A and Ad12ElB19K being expressed at early times. It was observed that 6fElA bound to CtBP very inefficiently. Ad12E1 transformed rat cell lines, carrying the 6f mutation were established from the 33°C transformants but failed to express the Ad12E1B54K protein. After a number of weeks in culture the cells developed a mesenchymal character; expression of proteins such as E-cadherin, P-cadherin and γ catenin was much reduced and expression of fibronection increased. These observations are consistent with inhibition of CtBP activity in wt Ad12E1 transformants but not in the 6f transformed cells. In a complementary study the effect of down-regulation of CtBP expression (using siRNA protocols) was examined. Consistent with results obtained with the 6f virus it was observed that reduction in expression of CtBP1 and CtBP2 facilitated viral infection and this effect was enhanced when expression of C-terminal interacting protein (CTIP) was also reduced.
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Grand, R.J.A. et al. (2007). The Significance of the CtBP — AdE1A Interaction during Viral Infection and Transformation. In: GtBP Family Proteins. Molecular Biology Intelligence Unit. Springer, New York, NY. https://doi.org/10.1007/978-0-387-39973-7_5
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DOI: https://doi.org/10.1007/978-0-387-39973-7_5
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