title: Human cytomegalovirus mtrII oncoprotein binds to p53 and down-regulates p53-activated transcription.
abstract: The 79-amino-acid (79-aa) open reading frame (UL111a) gene within morphological transforming region II (mtrII) of human cytomegalovirus strain Towne has been shown to transform rodent cells in vitro (J. Thompson, J. Doniger, and L. J. Rosenthal, Arch. Virol. 136:161-172, 1994). Moreover, a translation termination linker (TTL) mutant of mtrII that coded for the first 49 aa of mtrII oncoprotein (designated TTL49) was sufficient for malignant transformation, whereas a TTL mutant that coded for the first 24 aa (designated TTL24) was not. The current study demonstrates the binding of mtrII oncoprotein to the tumor suppressor protein p53 both in vivo using transiently transfected cells and in vitro using labeled proteins. Furthermore, the C-terminally truncated mtrII protein TTL49, but not truncated protein TTL24, bound to p53. The mtrII binding domain mapped to the N-terminal region of p53, residues 1 to 106, with a critical region from aa 27 to 44, whereas the p53 binding domain of mtrII protein was the first 49 aa. Furthermore, mtrII inhibited p53-activated transcription, indicating its ability to alter p53-directed cellular regulatory mechanisms. mtrII oncoprotein was detected both in stably transfected NIH 3T3 cell lines and human cytomegalovirus-infected HEL 299 cells (as early as 12 h after infection) in the perinuclear region and in the nucleus. mtrII-transformed cell lines, at both early and late passage, exhibited high levels of p53 with a 15-fold-extended half-life. However, p53-activated transcription was suppressed in these cells in spite of the increased p53 levels. Finally, the results with wild-type mtrII and its TTL mutants with respect to p53 binding, p53-activated transcription, and transforming ability suggest that the mechanism of mtrII transformation is linked to both p53 binding and disruption of p53 cell regulation.
authors.affiliation: Department of Microbiology and Immunology, Georgetown University Medical Center, Washington, D.C. 20007, USA.
authors.forename: S
authors.initials: S
authors.lastname: Muralidhar
authors.forename: J
authors.initials: J
authors.lastname: Doniger
authors.forename: E
authors.initials: E
authors.lastname: Mendelson
authors.forename: J C
authors.initials: JC
authors.lastname: Araujo
authors.forename: F
authors.initials: F
authors.lastname: Kashanchi
authors.forename: N
authors.initials: N
authors.lastname: Azumi
authors.forename: J N
authors.initials: JN
authors.lastname: Brady
authors.forename: L J
authors.initials: LJ
authors.lastname: Rosenthal
mesh_terms: D016475:3T3 Cells; D000818:Animals; D002461:Cell Line, Transformed; D002467:Cell Nucleus / Q000378:metabolism; D003587:Cytomegalovirus / Q000235:genetics* / Q000378:metabolism; D015536:Down-Regulation; D015967:Gene Expression Regulation, Viral; D006801:Humans; D019169:Jurkat Cells; D051379:Mice; D015513:Oncogene Proteins / Q000235:genetics* / Q000378:metabolism; D011817:Rabbits; D013997:Time Factors; D015533:Transcriptional Activation; D014162:Transfection; D016159:Tumor Suppressor Protein p53 / Q000235:genetics* / Q000378:metabolism
chemical_list: D015513:Oncogene Proteins; D016159:Tumor Suppressor Protein p53