Suppression of Lck protooncogene expression in murine somatic cell hybrids between T lymphoma cells and fibroblasts.
Somatic cell hybrids were obtained by cell fusions between Lck-positive EL4 mouse T lymphoma cells and Lck-negative B82 mouse fibroblasts of S194 mouse plasmacytoma cells to examine negative control of lck gene expression in the resulting hybrids. Western blot analysis using a monoclonal antibody against the Lck protein showed a marked decrease in p56lck expression in B82 x EL4 (BEL) hybrids. In contrast to BEL hybrids, the level of p56lck was not changed significantly in S194 x EL4 (SEL) hybrids and was approximately one-half of that seen in EL4 cells. Diminished expression of the Lck protein in BEL hybrids paralleled downregulation of lck mRNA, which was exclusively transcribed from the distal promoter in EL4 cells. It is unlikely that the suppression was simply a consequence of chromosome segregation critical for lck gene expression, since BEL hybrids retained the EL4-derived lck gene and most of the chromosomes from both parental cells. The results from treatment of BEL hybrids with actinomycin D or cycloheximide suggested that suppression of lck gene expression in the hybrids might not be due to posttranscriptional control. DNA methylation status in the lck distal promoter and the coding regions did not appear to correlate with the expression of the gene. Our results suggest that negative control of lck gene expression differs between fibroblasts and B cells, in that lck gene expression in T cells can be shut down by transfer of a putative repressor factor or factors in fibroblasts but not in B cells.
Oikawa
T
T
Department of Cell Genetics, Sasaki Institute, University of Tokyo, Japan.
Yamada
T
T
Kubota
Y
Y
Kondoh
N
N
Hitomi
Y
Y
Uchiumi
F
F
Yamamoto
T
T
D000818:Animals; D001483:Base Sequence; D003513:Cycloheximide / Q000494:pharmacology; D017931:DNA Primers / Q000737:chemistry; D003609:Dactinomycin / Q000494:pharmacology; D005347:Fibroblasts / Q000502:physiology; D015972:Gene Expression Regulation, Neoplastic; D006822:Hybrid Cells; D019860:Lymphocyte Specific Protein Tyrosine Kinase p56(lck); D008223:Lymphoma / Q000235:genetics*; D008745:Methylation; D051379:Mice; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic; D011500:Protein Synthesis Inhibitors / Q000494:pharmacology; D011518:Proto-Oncogene Proteins / Q000235:genetics*; D011519:Proto-Oncogenes; D012333:RNA, Messenger / Q000235:genetics; D014158:Transcription, Genetic / Q000187:drug effects
D017931:DNA Primers; D011500:Protein Synthesis Inhibitors; D011518:Proto-Oncogene Proteins; D012333:RNA, Messenger; D003609:Dactinomycin; D003513:Cycloheximide; D019860:Lymphocyte Specific Protein Tyrosine Kinase p56(lck)