Novel Drosophila tumor necrosis factor class molecule ("DmTNF") and variants thereof

Abstract

The present invention provides novel polynucleotides encoding Drosophila DmTNF polypeptides, fragments and homologs thereof. The present invention also is directed to novel polynucleotides encoding two Drosophila DmTNF variants, DmTNFv1 and DmTNFv2 polypeptides, fragments and homologs thereof. Also provided are vectors, host cells, antibodies, and recombinant and synthetic methods for producing said polypeptides. The invention further relates to screening methods for identifying agonists and antagonists of the polynucleotides and polypeptides of the present invention, in addition to methods of genetically modifying Drosophila or cultured cells to express or mis-express DmTNF, DmTNFv1, or DmTNFv2. The invention also relates to the use of such modified insects or cells to characterize DmTNF activity, identify TNF-like genes and/or genes implicated in modulating TNF, characterize TNF signaling pathways, and/or to identify modulators of DmTNF activity.

1 - 40 . (canceled)  
     
     
         41 . A transgenic  D. melanogaster  comprising a transgene having a polynucleotide sequence selected from the group consisting of: 
 a.) an isolated polynucleotide encoding a polypeptide comprising amino acids 1 to 409 of SEQ ID NO:6;    (b) an isolated polynucleotide encoding a polypeptide comprising amino acids 2 to 409 of SEQ ID NO:6;    (c) an isolated polynucleotide encoding a polypeptide comprising amino acids 53 to 409 of SEQ ID NO:6;    (d) An isolated polynucleotide encoding a polypeptide comprising amino acids 62 to 409 of SEQ ID NO:6; and    (d) An isolated polynucleotide encoding a polypeptide comprising at least 332 contiguous amino acids of the polypeptide provided as SEQ ID NO:6;    wherein said transgene is operably linked to a constitutive, regulatable, or tissue specific expression control element.    
     
     
         42 . The transgenic  D. melanogaster  according to  claim 41 , wherein elevated expression of said transgene results in apoptosis in tissues in which said transgene is expressed.  
     
     
         43 . The transgenic  D. melanogaster  according to  claim 41 , wherein said control element comprises UAS elements upstream of the coding region of said polynucleotide sequence.  
     
     
         44 . The transgenic  D. melanogaster  according to  claim 43 , further comprising a second transgene encoding the GAL4 transactivator in operable linkage with the eye-specific promoter, GMR.  
     
     
         45 . The transgenic  D. melanogaster  according to  claim 44 , wherein activation of said GMR promoter results in overexpression of said encoded polypeptide and a pro-apoptotic phenotype in the eye of said transgenic  D. melanogaster.    
     
     
         46 . A method of screening for an inhibitor of DmTNFv2 comprising: 
 (a) administering a candidate inhibitor to the transgenic  D. melanogaster  of either  claim 42  or  45 , and    (b) observing any reversion in phenotype to normal eye morphology in said D. melanogaster, wherein said reversion is indicative of DmTNFv2 inhibitor activity.    
     
     
         47 . A method of identifying a suppressor of DmTNFv2 comprising: 
 a.) mating the transgenic  D. melanogaster  of  claim 42  or  45  with a  D. melanogaster  containing a genomic deletion, and    b.) observing any reversion in phenotype to normal eye morphology in said genomic deletion  D. melanogaster,  wherein said reversion is indicative of the deleted genomic region containing a suppressor of DmTNFv2.