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Adenovirus type 5 E1A immortalizes primary rat cells expressing wild-type p53
T Yamashita1, H Tonoki, D Nakata
1Department of Molecular Biology, Cancer Research Institute, Sapporo Medical University School of Medicine, Hokkaido, Japan. yamasita@sapmed.ac.jp
Microbiology and Immunology
|December 28, 1999
Summary
Adenovirus E1A immortalization of rat cells does not require p53 loss. Wild-type p53 is functional in most Ad5 E1A-immortalized and spontaneously immortalized rat cell lines, suggesting p53 mutation is infrequent.
Area of Science:
- Molecular Biology
- Virology
- Cell Biology
Background:
- Adenovirus (Ad) E1A induces apoptosis in wild-type p53 cells.
- Stable transformation by Ad E1A necessitates co-introduction of anti-apoptotic genes like Ad E1B 19K.
- Cells immortalized by Ad E1A alone may have lost functional p53.
Purpose of the Study:
- To analyze the status and function of p53 in rat cells expressing Ad E1A.
- To investigate whether functional loss of p53 is a prerequisite for E1A-mediated immortalization.
Main Methods:
- Established rat cell lines by transfecting primary rat embryo fibroblast (REF) and baby rat kidney (BRK) cells with cloned Ad5 E1A.
- Utilized a yeast functional assay to analyze p53 in Ad5 E1A-immortalized and spontaneously immortalized rat cell lines.
- Detected p53 using immunoprecipitation in Ad5 E1A-immortalized cell lines.
Main Results:
- All analyzed spontaneously and Ad5 E1A-immortalized rat cell lines, except for 3Y1, expressed wild-type p53.
- All Ad5 E1A-immortalized rat cell lines showed detectable p53 via immunoprecipitation.
- Recombinant adenovirus expressing wild-type rat or murine p53 induced apoptosis in p53-null cells when collaborating with E1A.
Conclusions:
- p53 mutation is not frequent in spontaneous immortalization of primary rat cells.
- Functional loss of wild-type p53 is not a prerequisite for E1A-mediated immortalization.
- E1A can immortalize cells while maintaining functional wild-type p53.