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Mutations in polyomavirus middle T antigen affecting tumorigenesis.

C Gelinas1, B Schaffhausen, B Bockus

  • 1Department of Microbiology, University of Sherbrooke, Quebec, Canada.

Virology
|May 1, 1989
PubMed
Summary

A polyomavirus mutant (P155) transforms cells normally but shows reduced tumor formation. Specific mutations in middle T antigen impair tumorigenesis without affecting pp60c-src association or activation.

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Area of Science:

  • Virology
  • Molecular Biology
  • Oncogenesis

Background:

  • Polyomavirus middle T antigen is crucial for cell transformation and tumor development.
  • The P155 mutant exhibits normal transforming ability but diminished tumorigenic potential due to a specific deletion in middle T antigen.

Purpose of the Study:

  • To investigate the role of specific amino acid residues within the P155 deletion in middle T antigen's function.
  • To elucidate the biochemical mechanisms underlying the impaired tumorigenic potential of P155 mutants.

Main Methods:

  • Deletion loop mutagenesis was employed to introduce point mutations into the wild-type middle T antigen sequence.
  • Mutant middle T antigens were analyzed for their ability to transform cells in culture and induce tumors in rats.
  • Biochemical assays assessed middle T antigen association with and activation of cellular src kinase (pp60c-src) and its phosphorylation patterns.

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Main Results:

  • A single amino acid substitution (Ala to Val at residue 373) mimicked the P155 phenotype, impairing tumor induction while retaining transforming ability.
  • Mutant middle T antigens associated with and activated pp60c-src.
  • Mutant middle T antigens displayed slightly altered phosphorylation patterns compared to wild-type.

Conclusions:

  • The region encompassing amino acids 372-375 in polyomavirus middle T antigen is critical for tumorigenesis but not for cell transformation or pp60c-src activation.
  • Altered phosphorylation patterns of middle T antigen may contribute to its reduced tumorigenic potential.