j Human T-cell lymphotropic virus type 1 (HTLV-1)-induced adult T-cell leukemia/lymphoma is an aggressive malignancy. HTLV-2 is genetically related to HTLV-1 but does not cause any malignant disease. HTLV-1 Tax transactivator (Tax-1) contributes to leukemogenesis via NF-B. We describe transgenic Drosophila models expressing Tax in the compound eye and plasmatocytes. We demonstrate that Tax-1 but not Tax-2 induces ommatidial perturbation and increased plasmatocyte proliferation and that the eye phenotype is dependent on Kenny (IKK␥/NEMO), thus validating this new in vivo model. Adult T-cell leukemia/lymphoma (ATL) is an aggressive malignancy secondary to HTLV-1 (human T-cell lymphotropic virus type 1) infection (1). Although HTLV-1 and HTLV-2 are similar in genetic organization, they display major differences in pathogenesis and disease manifestation. HTLV-1 is capable of transforming T lymphocytes in infected individuals and subsequently leads to ATL, whereas HTLV-2 has not been clearly associated with malignant diseases but only with lymphocytosis (2, 3).Transgenic mouse models overexpressing Tax-1 demonstrate its oncogenic properties (4-6). However, cellular pathways and in vivo Tax-1 partners that mediate Tax-1-induced cellular transformation are still unexplored. Drosophila melanogaster is a valuable model because of the availability of facile genetic screens, a nearly complete collection of mutants, RNA interference (RNAi) lines, and advanced genetic technologies, in addition to highly conserved pathways such as NF-B. Acquisition of a "rough-eye" phenotype is an accepted surrogate for cell transformation in the Drosophila model. Using Drosophila transgenic models expressing Tax proteins (Tax-1 and Tax-2), we present evidence demonstrating the ability of Tax-1 but not Tax-2 to induce transformation in Drosophila and show that this transformation is dependent primarily on Kenny, the Drosophila orthologue of IKK␥/NEMO, upstream of Relish (NF-B) activation.Briefly, we overexpressed Tax-1 specifically in developing imaginal eyes by using the glass multimer reporter promoter GMRGal4 (number 9146; Bloomington Drosophila Stock Center, NIH P40OD018537). Tax-1 was amplified from pSG5M-Tax (7) and cloned into the Drosophila expression vector pUAST (GenScript) with an N-terminal Myc tag. Plasmid pUAST-Tax-1 was used for the generation of transgenic fly strains after injection into wildtype white-eyed (white Ϫ ) embryos (BestGene). Successful transgenesis was monitored through the appearance of the red-eye phenotype (white ϩ ). Analysis of the ommatidial structure by scanning electron microscopy was performed with a Tescan Mira3 LMU field emission gun scanning electron microscope. A grading system based on the severity of the eye phenotype (number of ommatidial fusions and extent of bristle organization) was developed, and statistical tests were done by one-way analysis of variance. While control flies displayed normal eyes (Fig. 1A, left panel), the eyes of adult flies expressing a single copy of Tax-1 under the contr...
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