title: Walleye dermal sarcoma virus retroviral cyclin directly contacts TAF9.
abstract: Walleye dermal sarcoma virus (WDSV) is a complex retrovirus associated with dermal sarcomas in walleye fish. A WDSV accessory gene encodes a cyclin homolog or retroviral cyclin (rv-cyclin). WDSV rv-cyclin was found to be associated with transcription complexes and to affect transcription in a cell-type and promoter-dependent manner. It inhibited the WDSV promoter in walleye fibroblasts and activated transcription from GAL4 promoters when fused to the GAL4 DNA binding domain, and an activation domain (AD) has been localized to 30 amino acids in the carboxyl region. rv-cyclin can block the pulldown of transcription coactivators by the AD of VP16, and the isolated rv-cyclin AD interferes specifically with the interaction between the carboxyl halves of the VP16 AD, VP16C, and TATA-binding protein-associated factor 9 (TAF9). The carboxyl region and isolated AD can bind TAF9 directly in assays of protein-protein interaction in vitro. Furthermore, rv-cyclin and the isolated rv-cyclin AD interfere specifically with the function of VP16C in transcription assays. A previously identified motif within the VP16C sequence mediates TAF9 binding, and this motif is present in the activation domains of a variety of TAF9-binding transcriptional activators. A similar motif is present in the rv-cyclin AD, and point mutations within this motif affect rv-cyclin function and protein-protein interactions. The results support a model of transcription regulation by direct interaction with TAF9.
authors.affiliation: Department of Microbiology, Immunology, and Pathology, Campus Delivery 1619, Colorado State University, Fort Collins, CO 80523, USA. sandra.quackenbush@colostate.edu
authors.forename: Joel
authors.initials: J
authors.lastname: Rovnak
authors.forename: Sandra L
authors.initials: SL
authors.lastname: Quackenbush
mesh_terms: D020816:Amino Acid Motifs / Q000235:genetics; D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D016213:Cyclins / Q000235:genetics / Q000378:metabolism*; D017931:DNA Primers; D030283:Epsilonretrovirus / Q000235:genetics*; D005982:Glutathione Transferase; D008156:Luciferases; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D011401:Promoter Regions, Genetic / Q000235:genetics; D017434:Protein Structure, Tertiary; D017422:Sequence Analysis, DNA; D035381:TATA-Binding Protein Associated Factors / Q000378:metabolism*; D014158:Transcription, Genetic / Q000235:genetics*; D014764:Viral Proteins / Q000378:metabolism*
chemical_list: D016213:Cyclins; D017931:DNA Primers; D035381:TATA-Binding Protein Associated Factors; D014764:Viral Proteins; D008156:Luciferases; D005982:Glutathione Transferase