Oncoviral bovine leukemia virus G4 and human T-cell leukemia virus type 1 p13(II) accessory proteins interact with farnesyl pyrophosphate synthetase.
G4 and p13(II) are accessory proteins encoded by the X region of bovine leukemia virus and human T-cell leukemia virus type 1 (HTLV-1), respectively. Disruption of the G4 and p13(II) open reading frames interferes with viral spread in animal model systems, indicating that the corresponding proteins play a key role in viral replication. In addition, G4 is oncogenic in primary cell cultures and is absolutely required for efficient onset of leukemogenesis in sheep. To gain insight into the function of these proteins, we utilized the yeast two-hybrid system to identify protein partners of G4. Results revealed that G4 interacts with farnesyl pyrophosphate synthetase (FPPS), a protein involved in the mevalonate/squalene pathway and in synthesis of FPP, a substrate required for prenylation of Ras. The specificity of the interaction was verified by glutathione S-transferase (GST) pull-down assays and by coimmunoprecipitation experiments. Furthermore, confocal microscopy showed that the subcellular localization of G4 was profoundly affected by FPPS. The G4 protein itself was not prenylated, at least in rabbit reticulocyte lysate-based assays. The domain of G4 required for binding to FPPS was restricted to an amphipathic alpha-helix rich in arginine residues. Subtle mutation of this alpha-helix abrogated G4 oncogenic potential in vitro, providing a biological relevance for FPPS-G4 complex formation in cells. Finally, HTLV-1 p13(II) was also found to specifically interact with FPPS (in yeast as well as in GST pull-down assays) and to colocalize with G4 in mitochondria, suggesting a functional analogy between these oncoviral accessory proteins. Identification of FPPS as a molecular partner for p13(II) and G4 accessory proteins opens new prospects for treatment of retrovirus-induced leukemia.
Lefèbvre
Laurent
L
Faculty of Agronomy, Gembloux, Belgium.
Vanderplasschen
Alain
A
Ciminale
Vincenzo
V
Heremans
Hubertine
H
Dangoisse
Olivier
O
Jauniaux
Jean-Claude
JC
Toussaint
Jean-François
JF
Zelnik
Vlado
V
Burny
Arsène
A
Kettmann
Richard
R
Willems
Luc
L
D019883:Alkyl and Aryl Transferases / Q000235:genetics / Q000378:metabolism*; D000595:Amino Acid Sequence; D000818:Animals; D001120:Arginine / Q000378:metabolism; D001665:Binding Sites; D001692:Biological Transport; D002417:Cattle; D002472:Cell Transformation, Viral; D003593:Cytoplasm / Q000378:metabolism; D016355:Genes, pX; D051228:Geranyltranstransferase; D006367:HeLa Cells; D015368:Human T-lymphotropic virus 1 / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D001909:Leukemia Virus, Bovine / Q000235:genetics / Q000378:metabolism*; D008024:Ligands; D008928:Mitochondria / Q000378:metabolism; D008969:Molecular Sequence Data; D009856:Oncogene Proteins, Viral / Q000235:genetics / Q000378:metabolism*; D012191:Retroviridae Proteins / Q000235:genetics / Q000378:metabolism*; D017386:Sequence Homology, Amino Acid; D018631:ras Proteins / Q000378:metabolism
D008024:Ligands; D009856:Oncogene Proteins, Viral; D012191:Retroviridae Proteins; C446341:g4 protein, Bovine leukemia virus; C072813:rof protein, Human T-lymphotropic virus 1; D001120:Arginine; D019883:Alkyl and Aryl Transferases; D051228:Geranyltranstransferase; D018631:ras Proteins