title: The nature of the catalytic domain of 2'-5'-oligoadenylate synthetases.
abstract: 2'-5'-Oligoadenylate (2-5(A)) synthetases are a family of interferon-induced enzymes that are activated by double-stranded RNA. To understand why, unlike other DNA and RNA polymerases, they catalyze 2'-5' instead of 3'-5' phosphodiester bond formation, we used molecular modeling to compare the structure of the catalytic domain of DNA polymerase beta (pol beta) to that of a region of the P69 isozyme of 2-5(A) synthetase. Although the primary sequence identity is low, like pol beta, P69 can assume an alphabetabetaalphabetabetabeta structure in this region. Moreover, mutation of the three Asp residues of P69, which correspond to the three catalytic site Asp residues of pol beta, inactivated the enzyme without affecting its substrate and activator binding capacity, providing further credence to the concept that this region is the catalytic domain of P69. This domain is highly conserved among all 2-5(A) synthetase isozymes. Biochemical and mutational studies demonstrated that dimerization of the P69 protein is required for its enzyme activity. However, a dimer containing a wild type subunit and an inactive catalytic domain mutant subunit was also active. The rate of catalysis of the heterodimer was half of that of the wild type homodimer, although the two proteins bound double-stranded RNA and ATP equally well.
authors.affiliation: Department of Molecular Biology, Lerner Research Institute, The Cleveland Clinic Foundation, Cleveland, Ohio 44195, USA.
authors.forename: S N
authors.initials: SN
authors.lastname: Sarkar
authors.forename: A
authors.initials: A
authors.lastname: Ghosh
authors.forename: H W
authors.initials: HW
authors.lastname: Wang
authors.forename: S S
authors.initials: SS
authors.lastname: Sung
authors.forename: G C
authors.initials: GC
authors.lastname: Sen
mesh_terms: D015088:2',5'-Oligoadenylate Synthetase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D000595:Amino Acid Sequence; D000818:Animals; D002384:Catalysis; D019281:Dimerization; D006801:Humans; D008969:Molecular Sequence Data; D009154:Mutation
chemical_list: D015088:2',5'-Oligoadenylate Synthetase