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Related Experiment Videos

A viable mouse polyomavirus mutant without immortalizing or transforming activities.

S Linder1, M Nilsson, I Martens

  • 1Department of Medical Virology, Uppsala University Biomedical Center, Sweden.

Virology
|November 1, 1990
PubMed
Summary

A polyomavirus mutant (dl1041) with a large deletion in its T-antigens shows defective immortalization and transformation. However, it retains DNA replication and late promoter activation capabilities, indicating distinct functions for T-antigens.

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Area of Science:

  • Virology
  • Molecular Biology
  • Oncology

Background:

  • Polyomaviruses encode large and middle T-antigens crucial for viral replication and host cell manipulation.
  • Mutants with deletions in T-antigen genes provide insights into their specific functions.

Purpose of the Study:

  • To characterize the polyomavirus mutant dl1041, which has a deletion affecting T-antigen coding sequences.
  • To investigate the roles of large and middle T-antigens in viral DNA replication, cell immortalization, transformation, and promoter activation.

Main Methods:

  • Construction and characterization of the polyomavirus mutant dl1041 with a 375-base pair deletion.
  • Conditional viability assays in different mouse cell lines (3T3 and 3T6).
  • DNA replication, transformation, and viral late promoter trans-activation assays.

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  • Analysis of T-antigen fragments for immortalizing activity.

    • Main Results:

    • The dl1041 mutant exhibited conditional viability with defective immortalizing and transforming functions.
    • dl1041 replicated its DNA in rapidly growing 3T6 cells but not efficiently in serum-deprived 3T3 cells.
    • The mutant completed an infection cycle, producing infectious virus in 3T6 cells.
    • Large T-antigen of dl1041 retained the ability to activate the viral late promoter in trans.
    • A specific region (amino acid residues 136-184) of the large T-antigen was identified as essential for immortalizing activity.

    Conclusions:

    • Immortalization and viral late promoter trans-activation are distinct functions of the polyomavirus large T-antigen.
    • The dl1041 mutant provides a tool to dissect the complex functions of T-antigens.
    • Cellular growth state influences the efficiency of viral DNA replication, potentially linked to T-antigen deficiencies.