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Immunologically distinct p53 molecules generated by alternative splicing.

N Arai, D Nomura, K Yokota

    Molecular and Cellular Biology
    |September 1, 1986
    PubMed
    Summary
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    Researchers reconstituted p53 expression in L12 cells using a functional p53 gene. Variations in p53 protein epitopes were linked to alternative splicing in cDNA clones, affecting protein length and antigenic sites.

    Area of Science:

    • Molecular Biology
    • Cancer Research
    • Genetics

    Background:

    • The tumor suppressor protein p53 plays a critical role in cellular responses to DNA damage.
    • Understanding p53 expression and its variations is crucial for cancer research.
    • Previous studies reconstituted p53 expression in nonproducer cell lines.

    Purpose of the Study:

    • To compare the immunological epitopes of p53 proteins encoded by different full-length cDNA clones.
    • To investigate the molecular basis for observed differences in p53 protein expression and immunological profiles.
    • To determine if alternative splicing mechanisms contribute to p53 mRNA diversity in vivo.

    Main Methods:

    • Transfection of p53 cDNA clones into L12 cells.
    • Immunoprecipitation and in vitro transcription/translation of p53 proteins.

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  • DNA sequence analysis of p53 cDNA clones.
  • Differential hybridization of cellular mRNA populations.

    • Main Results:

    • Reconstitution of p53 expression in L12 cells resulted in proteins indistinguishable from naturally occurring tumor cell p53.
    • Different p53 cDNA clones yielded proteins with variations in immunological epitopes, specifically the PAb421-PAb122 site.
    • Sequence analysis revealed a 96-bp insert in one clone (p53-M8) containing a premature termination signal, resulting in a truncated p53 protein.
    • This insert was homologous to p53 intron 10 sequences, suggesting alternative splicing.

    Conclusions:

    • The distinct immunological profiles of p53 proteins are due to alternative splicing of p53 mRNA.
    • Alternative splicing generates different p53 mRNA species, leading to variations in protein length and epitope exposure.
    • These alternatively spliced p53 mRNA species are expressed in vivo in transformed cells.