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Yeast As a Chassis for Developing Functional Assays to Study Human P53
Published on: August 4, 2019
The presence of the p53 transcripts with truncated open reading frames in Marek's disease tumor-derived cell lines
Michihiro Takagi1, Kazuhiko Ohashi, Toshifumi Morimura
1Department of Microbiology and Immunology, Faculty of Agriculture, Kobe University, Kobe 657-8501, Japan.
Abstract:
Several kinds of the p53 transcripts in which their open reading frames (ORFs) were truncated (ranging from 101 to 765 bp) were identified in Marek's disease (MD)-derived tumor cell lines as well as avian leukosis- and reticuloendotheliosis-derived ones, detected by nested RT-PCR and subsequent nucleotide sequence analysis. In these ORFs, regions encoding the proline-rich and DNA-binding domains of the p53 protein were frequently deleted, and many of these deletions were found to cause frame shift. Western blot analysis using anti-p53 monoclonal antibodies revealed that multiple p53 isoform proteins with various molecular weights including 45-46, 35 and 28 kDa were expressed in these tumor cell lines, though the p53 protein with a molecular weight of 49 kDa was detected in chicken embryo fibroblasts transformed by the SV40 T antigen as a control. Since no deletions were found in the p53 gene of these MD tumor cell lines, truncations in the p53 ORFs observed in this study might result from alternative splicing of the p53 gene.
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