title: Enhanced malignant transformation induced by expression of a distinct protein domain of ribonucleotide reductase large subunit from herpes simplex virus type 2.
abstract: The 1.3-kilobase (kb) Pst I DNA fragment C (Pst I-C) of herpes simplex virus type 2 (HSV-2) morphological transforming region III (mtrIII; map unit 0.562-0.570) encodes part of the N-terminal half of the large subunit of ribonucleotide reductase (RR1; amino acid residues 71-502) and induces the neoplastic transformation of immortalized cell lines. To assess directly the role of these RR1 protein sequences in cell transformation, the Pst I-C fragment was cloned in an expression vector (p91023) containing an adenovirus-simian virus 40 promoter-enhancer to generate recombinant plasmid p9-C. Expression of a protein domain (approximately 65 kDa) was observed in p9-C-transfected COS-7 and Rat2 cells but not in those transfected with plasmid pHC-14 (Pst I-C in a promoterless vector). In Rat2 cells, p9-C induced highly transformed foci at an elevated frequency compared with that of pHC-14. Introduction of translation termination (TAG) condons within the RR1 coding sequence and within all three reading frames inactivated RR1 protein expression from p9-C and reduced its transforming activity to the level seen with the standard pHC-14 construct. Wild-type p9-C specified a protein kinase capable of autophosphorylation. Computer-assisted analysis further revealed significant similarity between regions of mtrIII-specific RR1 and amino acid patterns conserved within the proinsulin precursor family and DNA transposition proteins. These results identify a distinct domain of the HSV-2 RR1 protein involved in the induction of enhanced malignant transformation. In addition, the data indicate that the mtrIII DNA itself can induce basal-level transformation in the absence of protein expression.
authors.affiliation: Viral Carcinogenesis Laboratories, Linus Pauling Institute of Science and Medicine, Palo Alto, CA 94306.
authors.forename: M A
authors.initials: MA
authors.lastname: Ali
authors.forename: D
authors.initials: D
authors.lastname: McWeeney
authors.forename: A
authors.initials: A
authors.lastname: Milosavljevic
authors.forename: J
authors.initials: J
authors.lastname: Jurka
authors.forename: R J
authors.initials: RJ
authors.lastname: Jariwalla
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002472:Cell Transformation, Viral; D002478:Cells, Cultured; D002522:Chlorocebus aethiops; D003001:Cloning, Molecular; D004252:DNA Mutational Analysis; D004279:DNA, Viral / Q000235:genetics; D005814:Genes, Viral; D046911:Macromolecular Substances; D008969:Molecular Sequence Data; D009857:Oncogenes; D011494:Protein Kinases / Q000235:genetics / Q000378:metabolism; D051381:Rats; D012264:Ribonucleotide Reductases / Q000737:chemistry / Q000276:immunology / Q000502:physiology*; D016415:Sequence Alignment; D018139:Simplexvirus / Q000201:enzymology*; D015678:Viral Structural Proteins / Q000235:genetics
chemical_list: D004279:DNA, Viral; D046911:Macromolecular Substances; D015678:Viral Structural Proteins; D012264:Ribonucleotide Reductases; D011494:Protein Kinases