title: Characterization of human phosphodiesterase 12 and identification of a novel 2'-5' oligoadenylate nuclease - The ectonucleotide pyrophosphatase/phosphodiesterase 1.
abstract: The vertebrate 2-5A system is part of the innate immune response and central to cellular antiviral activities. Upon activation by viral double-stranded RNA, 5'-triphosphorylated, 2'-5'-linked oligoadenylate polyribonucleotides (2-5As) are synthesized by one of several 2'-5' oligoadenylate synthetases. The 2-5As bind and activate RNase L, an unspecific endoribonuclease, resulting in viral and cellular RNA decay. Given that most endogenous RNAs are degraded by RNase L, continued enzyme activity will eventually lead to cell growth arrest and cell death. This is averted, when 2-5As and their 5'-dephosphorylated forms, the so-called 2-5A core molecules, are cleaved and thus inactivated by 2'-5'-specific nuclease(s), e.g. phosphodiesterase 12, thereby turning RNase L into its latent form. In this study, we have characterized the human phosphodiesterase 12 in vitro focusing on its ability to degrade 2-5As and 2-5A core molecules. We have found that the enzyme activity is distributive and is influenced by temperature, pH and divalent cations. This allowed us to determine V(max) and K(m) kinetic parameters for the enzyme. We have also identified a novel 2'-5'-oligoadenylate nuclease; the human plasma membrane-bound ectonucleotide pyrophosphatase/phosphodiesterase 1, suggesting that 2-5A catabolism may be a multienzyme-regulated process.
authors.affiliation: Department of Molecular Biology and Genetics, Aarhus University, C. F. Møllers Allé 3, DK-8000 Aarhus C, Denmark. jbp@mb.au.dk
authors.forename: Jesper B
authors.initials: JB
authors.lastname: Poulsen
authors.forename: Kasper R
authors.initials: KR
authors.lastname: Andersen
authors.forename: Karina H
authors.initials: KH
authors.lastname: Kjær
authors.forename: Anna L
authors.initials: AL
authors.lastname: Vestergaard
authors.forename: Just
authors.initials: J
authors.lastname: Justesen
authors.forename: Pia M
authors.initials: PM
authors.lastname: Martensen
mesh_terms: D000227:Adenine Nucleotides / Q000378:metabolism*; D019295:Computational Biology; D006801:Humans; D015151:Immunoblotting; D009843:Oligoribonucleotides / Q000378:metabolism*; D010727:Phosphoric Diester Hydrolases / Q000235:genetics / Q000378:metabolism*; D011755:Pyrophosphatases / Q000235:genetics / Q000378:metabolism*; D012260:Ribonucleases / Q000235:genetics / Q000378:metabolism*
chemical_list: D000227:Adenine Nucleotides; D009843:Oligoribonucleotides; C023505:2',5'-oligoadenylate; D012260:Ribonucleases; D010727:Phosphoric Diester Hydrolases; C065962:ectonucleotide pyrophosphatase phosphodiesterase 1; D011755:Pyrophosphatases