Perturbation of TSG101 protein affects cell cycle progression.
tsg101 was recently identified as a tumor susceptibility gene by functional inactivation of allelic loci in mouse 3T3 fibroblasts. Although previous studies suggested that homozygous intragenic deletion of TSG101 is rare in breast cancer cells and specimens, the neoplastic phenotype caused by tsg101 inactivation implicated that tsg101 may play a significant role in cell growth control. Here, we characterize mouse polyclonal and monoclonal antibodies that specifically recognize the TSG101 protein (molecular mass, 46 kDa) in whole-cell lysates by straight Western blot analysis. By indirect immunofluorescence staining, TSG101 was found to be localized in the cytoplasm throughout the entire cell cycle. However, the nuclear staining increases from G1 to S phase and becomes dominant in late S phase. TSG101 is mainly distributed surrounding the chromosomes during M phase. The expression level of TSG101 is not cell cycle dependent. It is possible that the relocalization of TSG101 from the cytoplasm into the nucleus may be relevant to its function. Microinjection of both polyclonal and monoclonal antibodies specific to TSG101 into cells during G1 or S phase results in cell cycle arrest. Furthermore, overexpression of TSG101 leads to cell death, suggesting that the appropriate amount of TSG101 is critical for cell cycle progression. Taken together, these results suggest that neoplastic transformation caused by TSG101 deficiency may result from bypassing of the cell cycle checkpoints.
Zhong
Q
Q
Department of Molecular Medicine and Institute of Biotechnology, University of Texas Health Science Center at San Antonio, 78245, USA.
Chen
Y
Y
Jones
D
D
Lee
W H
WH
D000911:Antibodies, Monoclonal / Q000494:pharmacology; D016923:Cell Death; D002467:Cell Nucleus / Q000378:metabolism; D003593:Cytoplasm / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D056827:Endosomal Sorting Complexes Required for Transport; D016193:G1 Phase; D006801:Humans; D009363:Neoplasm Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D016196:S Phase; D014157:Transcription Factors / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D014407:Tumor Cells, Cultured; D001749:Urinary Bladder Neoplasms / Q000378:metabolism / Q000473:pathology; D000099177:Tsg101 Protein
D000911:Antibodies, Monoclonal; D004268:DNA-Binding Proteins; D056827:Endosomal Sorting Complexes Required for Transport; D009363:Neoplasm Proteins; D014157:Transcription Factors; D000099177:Tsg101 Protein