Targeted deletion of the Tsg101 gene results in cell cycle arrest at G1/S and p53-independent cell death

Andrea Krempler, Malinda D. Henry, Aleata A. Triplett, Kay Uwe Wagner

Research output: Contribution to journalArticle

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Abstract

The tumor susceptibility gene 101 (Tsg101) was originally discovered in a screen for potential tumor suppressors using insertional mutagenesis in immortalized fibroblasts. To investigate essential functions of this gene in cell growth and neoplastic transformation, we derived primary mouse embryonic fibroblasts from Tsg101 conditional knockout mice. Expression of Cre recombinase from a retroviral vector efficiently down-regulated Tsg101. The deletion of Tsg101 caused growth arrest and cell death but did not result in increased proliferation and cellular transformation. Inactivation of p53 had no influence on the deleterious phenotype, but Tsg101-/- cells were rescued through expression of exogenous Tsg101. Fluorescence-activated cell sorting, proliferation assays, and Western blot analysis of crucial regulators of the cell cycle revealed that Tsg101 deficiency resulted in growth arrest at the G1/S transition through inactivation of cyclin-dependent kinase 2. As a consequence, DNA replication was not initiated in Tsg101-deficient cells. Our results clearly demonstrate that Tsg101 is not a primary tumor suppressor in mouse embryonic fibroblasts. However, the protein is crucial for cell proliferation and cell survival.

Original languageEnglish (US)
Pages (from-to)43216-43223
Number of pages8
JournalJournal of Biological Chemistry
Volume277
Issue number45
DOIs
StatePublished - Nov 8 2002

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G1 Phase Cell Cycle Checkpoints
Cell death
Cell Death
Genes
Cells
Fibroblasts
Cell Proliferation
Tumors
Growth
Neoplastic Cell Transformation
Cyclin-Dependent Kinase 2
Tsg101 protein
Mutagenesis
Insertional Mutagenesis
Essential Genes
Cell proliferation
Cell growth
DNA Replication
Sorting
Knockout Mice

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Targeted deletion of the Tsg101 gene results in cell cycle arrest at G1/S and p53-independent cell death. / Krempler, Andrea; Henry, Malinda D.; Triplett, Aleata A.; Wagner, Kay Uwe.

In: Journal of Biological Chemistry, Vol. 277, No. 45, 08.11.2002, p. 43216-43223.

Research output: Contribution to journalArticle

Krempler, Andrea ; Henry, Malinda D. ; Triplett, Aleata A. ; Wagner, Kay Uwe. / Targeted deletion of the Tsg101 gene results in cell cycle arrest at G1/S and p53-independent cell death. In: Journal of Biological Chemistry. 2002 ; Vol. 277, No. 45. pp. 43216-43223.
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