Mutational analysis of the v-Rel dimerization interface reveals a critical role for v-Rel homodimers in transformation.
The v-rel oncogene encoded by reticuloendotheliosis virus strain T is the acutely transforming member of the Rel/NF-kappaB family of transcription factors. In v-Rel-transformed cells, v-Rel exists as homodimers or heterodimers with the endogenous Rel/NF-kappaB proteins c-Rel, NF-kappaB1, NF-kappaB2, and RelA. To examine the contribution of these complexes to v-Rel-mediated transformation, mutations were introduced into the dimerization interface of v-Rel to generate v-Rel mutants with selective dimerization properties. Nine mutants are described in this study that are defective in homodimer and/or heterodimer formation with specific Rel/NF-kappaB family members. Viruses expressing mutants that failed to homodimerize but were able to form heterodimeric complexes were unable to transform splenic lymphocytes in vitro, indicating that the dimerization of v-Rel with endogenously expressed Rel/NF-kappaB proteins is not in itself sufficient for transformation. In addition, two partially transforming mutants were identified that exhibited an impaired ability to form homodimers. Sequence analysis of the proviral DNA from cells transformed by these mutants revealed the presence of multiple secondary mutations in sequences responsible for dimerization and DNA binding. Two of these mutations either enhanced or restored the ability of these proteins to bind DNA as a homodimer. Viruses expressing these proteins transformed cells at levels comparable to or slightly less than v-Rel, suggesting that a threshold level of DNA binding by v-Rel homodimers is required for transformation.
Liss
Andrew S
AS
Section of Molecular Genetics and Microbiology, University of Texas at Austin, Austin, Texas 78712-1095, USA.
Bose
Henry R
HR
D000595:Amino Acid Sequence; D000818:Animals; D002472:Cell Transformation, Viral; D002642:Chick Embryo; D004252:DNA Mutational Analysis; D004279:DNA, Viral / Q000378:metabolism; D019281:Dimerization; D008214:Lymphocytes / Q000378:metabolism / Q000821:virology; D008969:Molecular Sequence Data; D016328:NF-kappa B / Q000378:metabolism; D020572:Oncogene Proteins v-rel / Q000235:genetics* / Q000378:metabolism; D011533:Proviruses / Q000235:genetics*; D001356:Reticuloendotheliosis virus / Q000737:chemistry / Q000235:genetics*
D004279:DNA, Viral; D016328:NF-kappa B; D020572:Oncogene Proteins v-rel