RESUMO
Murine retrovirus SL3-3 is highly T lymphomagenic. Its pathogenic properties are determined by the transcriptional enhancer of the U3 repeat region which shows preferential activity in T cells. Within the U3 repeats, the major determinant of T-cell specificity has been mapped to binding sites for the AML1 transcription factor family (also known as the core binding factor [CBF], polyomavirus enhancer binding protein 2 [PEBP2], and SL3-3 enhancer factor 1 [SEF-1]). SL3-3 viruses with AML1 site mutations have lost a major determinant of T-cell-specific enhancer function but have been found to retain a lymphomagenic potential, although disease induction is slower than for the SL3-3 wild type. To compare the specificities and mechanisms of disease induction of wild-type and mutant viruses, we have examined lymphomas induced by mutant viruses harboring transversions of three consecutive base pairs critical to AML1 site function (B. Hallberg, J. Schmidt, A. Luz, F. S. Pedersen, and T. Grundström. J. Virol. 65:4177-4181, 1991). Our results show that the mutated AML1 sites are genetically stable during lymphomagenesis and that ecotropic provirus numbers in DNA of tumors induced by wild-type and mutant viruses fall within the same range. Moreover, proviruses were found to be integrated at the c-myc locus in similar proportions of wild-type and mutant SL3-3-induced tumors, and the mutated AML1 sites of proviruses at c-myc are unaltered. In some cases, however, including one c-myc-integrated provirus, a single-base pair change was detected in a second, weaker AML1 binding site. By DNA rearrangement analysis of the T-cell receptor beta-locus, tumors induced by the AML1 site mutants are found to be of the T-cell type. Thus, although the AML1 site mutants have weakened T-cell-specific enhancers they are T-lymphomagenic, and wild-type- and mutant-virus-induced tumor DNAs are similar with respect to the number of overall ecotropic and c-myc-integrated clonal proviruses. The SL3-3 wild-type and AML1 site mutant viruses may therefore induce disease by similar mechanisms.
Assuntos
DNA de Neoplasias/análise , Proteínas de Ligação a DNA , Elementos Facilitadores Genéticos , Vírus da Leucemia Murina/genética , Leucemia Experimental/genética , Linfoma de Células T/genética , Proteínas Proto-Oncogênicas , Infecções por Retroviridae/genética , Fatores de Transcrição/genética , Infecções Tumorais por Vírus/genética , Animais , Sequência de Bases , Subunidade alfa 2 de Fator de Ligação ao Core , Humanos , Camundongos , Dados de Sequência Molecular , Mutação , Proteínas Proto-Oncogênicas c-myc/genética , Provírus/genética , Vacinas Atenuadas , Integração ViralRESUMO
The murine retrovirus SL3-3 is a potent inducer of T-cell lymphomas when inoculated into susceptible newborn mice. The proviral integration site sequences were surveyed in tumor DNAs by a simple two-step PCR method. From 20 SL3-3-induced tumors a total of 39 provirus-host junctions were amplified and sequenced. Seven showed homology to known sequences. These included the known common integration site c-myc as well as genes not previously identified as targets of provirus integration, namely N-ras and the genes coding for major histocompatibility complex class 11 E-beta, protein kinase C-eta, and T-cell receptor beta-chain. Among these genes, the integrations in c-myc as well as the one in N-ras were found to be clonal. One of the remaining 32 proviral integration site sequences that show no similarities to known sequences may represent a common integration site, as 2 of the 20 tumors demonstrated clonal provirus insertion into this region.