Characterization of the Cyno-EBV LMP1 homologue and comparison with LMP1s of EBV and other EBV-like viruses.
EBV latent membrane protein 1 (LMP1) is essential for EBV-mediated transformation and has been associated with several cases of malignancies. EBV-like viruses in Cynomolgus monkeys (Macaca fascicularis) have been associated with high lymphoma rates in immunosuppressed monkeys. In the study, the entire coding region of the Cyno-EBV LMP1 gene was cloned, sequenced and expressed in human embryonic kidney (HEK) cells 293. The Cyno-EBV LMP1 homologue sequence predicted a 588 amino acid (a.a.) protein with a short 19 a.a. N-terminus, six transmembrane domains and a long carboxy tail of 404 a.a. The protein contained a series of seven 9 a.a.-tandem repeats and two 20 a.a.-repeats, which harbored two potential TRAF binding motifs, PxQxT/S. These repeats shared no homology with the repeats in any other LMP1. However, the proline-rich sequence GPxxPx(6) found within the 11 a.a.-repeats of EBV LMP1 was conserved in Cyno-EBV carboxy tail and contained two consensus JAK/STAT sequences PxxPxP. A cluster of eight histidine residues was found in proximity to the last transmembrane domain of Cyno-EBV LMP1 and was exploited as a natural protein tag in expression studies. Western blot analysis revealed a major polypeptide of 110 kDa. Comparative functional studies showed that Cyno-EBV LMP1 expressed in HEK 293 cells shares the same ability as EBV LMP1 to induce NFkappaB driven CAT activity.
Faucher
Sylvie
S
Department of Biochemistry, Microbiology and Immunology, University of Ottawa, 451 Smyth Road, Ottawa Ont, Canada K1H 8M5. sylvie_faucher@hc-sc.gc.ca
Dimock
Kenneth
K
Wright
Kathryn E
KE
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D002472:Cell Transformation, Viral; D003001:Cloning, Molecular; D004854:Herpesvirus 4, Human / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D008252:Macaca fascicularis; D008969:Molecular Sequence Data; D016328:NF-kappa B / Q000378:metabolism; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D014162:Transfection; D014763:Viral Matrix Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism
C037662:EBV-associated membrane antigen, Epstein-Barr virus; D016328:NF-kappa B; D014763:Viral Matrix Proteins