Microsatellite instability in IVS3 of murine c-fes gene: tumor-associated rearrangement and mammalian divergence.
The murine lymphomacrophage hybrids ESb, EbF1, EbF2-c4, which express c-fes constitutively, were found by Southern analysis to bear a c-fes deletion of almost 100 bp. The deleted allele was transmitted to the metastatic hybrids by their nonexpressing, poorly metastatic T-lymphoma progenitor Eb, which also has a structurally normal c-fes allele. PCR amplification and sequencing of fes cDNA spanning exons 3-5, where the deletion mapped, ruled out any involvement of coding sequences in the rearrangement. PCR amplification of the as yet unsequenced murine c-fes IVS3 and IVS4 showed they are about 50% longer than their human and feline homologs. Sequencing of IVS4 showed no difference between tumor and control DNA. Sequencing of part of the approximately 2600-bp IVS3 was guided by the restriction analysis of PCR products from control and hybrid DNAs. This showed that differences from the control appeared to be mainly located in the 900-bp HindIII-EcoRI fragment, localized in the middle of IVS3. As all three hybrids had the same restriction map, this fragment was sequenced in one of them (ESb). A run of >200 CA repeats was found in control DNA, and a reduction in the CAn microsatellite accounted for most of the c-fes deletion in the ESb hybrid. Interestingly, the 50% reduction in the size of human and feline c-fes IVS3 as compared with the murine homolog is mostly due to contraction of the same microsatellite.
Volpi
L
L
Department of Biology and Genetics, Via Viotti 3/5, Medical Faculty, University of Milan, 20133 Milan, Italy.
Beghini
A
A
Rossi
D
D
Larizza
L
L
D000818:Animals; D001483:Base Sequence; D015139:Blotting, Southern; D002415:Cats; D002460:Cell Line; D017931:DNA Primers; D004276:DNA, Satellite / Q000235:genetics*; D005091:Exons; D006801:Humans; D016399:Lymphoma, T-Cell / Q000235:genetics; D051379:Mice; D018895:Microsatellite Repeats / Q000235:genetics*; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D011505:Protein-Tyrosine Kinases / Q000235:genetics*; D011518:Proto-Oncogene Proteins / Q000235:genetics*; D051578:Proto-Oncogene Proteins c-fes; D011519:Proto-Oncogenes; D015183:Restriction Mapping; D017384:Sequence Deletion; D012689:Sequence Homology, Nucleic Acid; D013045:Species Specificity; D014407:Tumor Cells, Cultured
D017931:DNA Primers; D004276:DNA, Satellite; D011518:Proto-Oncogene Proteins; D011505:Protein-Tyrosine Kinases; C496645:FES protein, human; C496646:Fes protein, mouse; D051578:Proto-Oncogene Proteins c-fes