Tsg101: a novel tumor susceptibility gene isolated by controlled homozygous functional knockout of allelic loci in mammalian cells.
Using a novel strategy that enables the isolation of previously unknown genes encoding selectable recessive phenotypes, we identified a gene (tsg101) whose homozygous functional disruption produces cell transformation. Antisense RNA from a transactivated promoter introduced randomly into transcribed genes throughout the genome of mouse 3T3 fibroblasts was used to knock out alleles of chromosomal genes adjacent to promoter inserts, generating clones that grew in 0.5% agar and formed metastatic tumors in nude mice. Removal of the transactivator restored normal growth. The protein encoded by tsg101 cDNA encodes a coiled-coil domain that interacts with stathmin, a cytosolic phosphoprotein implicated previously in tumorigenesis. Overexpression of tsg101 antisense transcripts in naive 3T3 cells resulted in cell transformation and increased stathmin-specific mRNA.
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Department of Genetics, Stanford University School of Medicine, California 94305-5120, USA.
Cohen
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D016475:3T3 Cells / Q000502:physiology; D000362:Agar; D000483:Alleles; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002454:Cell Differentiation / Q000235:genetics; D002455:Cell Division / Q000235:genetics; D002471:Cell Transformation, Neoplastic / Q000235:genetics; D002874:Chromosome Mapping; D003001:Cloning, Molecular; D003600:Cytosol / Q000502:physiology; D018076:DNA, Complementary / Q000235:genetics; D004268:DNA-Binding Proteins / Q000235:genetics*; D056827:Endosomal Sorting Complexes Required for Transport; D015972:Gene Expression Regulation, Neoplastic / Q000235:genetics; D005808:Genes, Recessive / Q000235:genetics; D016147:Genes, Tumor Suppressor / Q000235:genetics*; D006720:Homozygote; D008322:Mammals; D051379:Mice; D018345:Mice, Knockout; D008819:Mice, Nude; D008868:Microtubule Proteins; D008969:Molecular Sequence Data; D009374:Neoplasms, Experimental / Q000235:genetics* / Q000556:secondary; D010641:Phenotype; D010750:Phosphoproteins / Q000235:genetics; D016372:RNA, Antisense / Q000235:genetics; D012333:RNA, Messenger / Q000378:metabolism; D017422:Sequence Analysis, DNA; D050777:Stathmin; D014157:Transcription Factors / Q000235:genetics*; D014170:Transformation, Genetic; D000099177:Tsg101 Protein
D000362:Agar; D018076:DNA, Complementary; D004268:DNA-Binding Proteins; D056827:Endosomal Sorting Complexes Required for Transport; D008868:Microtubule Proteins; D010750:Phosphoproteins; D016372:RNA, Antisense; D012333:RNA, Messenger; D050777:Stathmin; D014157:Transcription Factors; D000099177:Tsg101 Protein; C493994:Stmn1 protein, mouse