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Related Experiment Videos

Type B leukemogenic virus has a T-cell-specific enhancer that binds AML-1.

J A Mertz1, F Mustafa, S Meyers

  • 1Section of Molecular Genetics and Microbiology and Institute for Cellular and Molecular Biology, The University of Texas at Austin, Austin, Texas 78712, USA.

Journal of Virology
|February 13, 2001
PubMed
Summary
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The Type B leukemogenic virus (TBLV) enhancer relies on AML-1 (RUNX1) binding for T-cell tumor development. This viral enhancer activity is cell-type specific and crucial for leukemogenesis.

Area of Science:

  • Virology
  • Molecular Biology
  • Oncology

Background:

  • Type B leukemogenic virus (TBLV) causes T-cell tumors in mice.
  • TBLV's long terminal repeats (LTRs) differ from MMTV due to deletions and triplications.
  • The LTR triplication may function as a viral enhancer element.

Purpose of the Study:

  • To investigate if the TBLV LTR triplication acts as a viral enhancer.
  • To identify transcription factors responsible for TBLV enhancer activity.
  • To understand the role of these factors in T-cell leukemogenesis.

Main Methods:

  • Insertion of the TBLV LTR triplication into a reporter construct (luciferase).
  • Reporter gene assays in various cell lines (T-cells, fibroblasts, B-cells, mammary cells).

Related Experiment Videos

  • Site-directed mutagenesis of the MMTV LTR to mimic TBLV deletion.
  • Gel shift assays to analyze transcription factor binding (AML-1, NF-A, NF-B).
  • Overexpression studies of AML-1 in mammary cell lines.

    • Main Results:

    • The TBLV LTR triplication demonstrated enhancer activity, particularly in T-cells.
    • Enhancer function was orientation-independent and cell-type specific.
    • A specific mutation (556M) significantly reduced TBLV enhancer activity (20-fold).
    • This mutation diminished binding of AML-1 (RUNX1) and other factors (NF-A, NF-B).
    • AML-1 overexpression increased TBLV LTR expression (30-fold) in mammary cells.

    Conclusions:

    • The TBLV LTR triplication functions as a cell-type specific enhancer, primarily in T-cells.
    • AML-1 (RUNX1) binding is essential for optimal TBLV enhancer activity.
    • AML-1, potentially with other factors, plays a critical role in TBLV-mediated leukemogenesis.