title: The Role of Phosphodiesterase 12 (PDE12) as a Negative Regulator of the Innate Immune Response and the Discovery of Antiviral Inhibitors.
abstract: 2',5'-Oligoadenylate synthetase (OAS) enzymes and RNase-L constitute a major effector arm of interferon (IFN)-mediated antiviral defense. OAS produces a unique oligonucleotide second messenger, 2',5'-oligoadenylate (2-5A), that binds and activates RNase-L. This pathway is down-regulated by virus- and host-encoded enzymes that degrade 2-5A. Phosphodiesterase 12 (PDE12) was the first cellular 2-5A- degrading enzyme to be purified and described at a molecular level. Inhibition of PDE12 may up-regulate the OAS/RNase-L pathway in response to viral infection resulting in increased resistance to a variety of viral pathogens. We generated a PDE12-null cell line, HeLaΔPDE12, using transcription activator-like effector nuclease-mediated gene inactivation. This cell line has increased 2-5A levels in response to IFN and poly(I-C), a double-stranded RNA mimic compared with the parental cell line. Moreover, HeLaΔPDE12 cells were resistant to viral pathogens, including encephalomyocarditis virus, human rhinovirus, and respiratory syncytial virus. Based on these results, we used DNA-encoded chemical library screening to identify starting points for inhibitor lead optimization. Compounds derived from this effort raise 2-5A levels and exhibit antiviral activity comparable with the effects observed with PDE12 gene inactivation. The crystal structure of PDE12 complexed with an inhibitor was solved providing insights into the structure-activity relationships of inhibitor potency and selectivity.
authors.affiliation: From the Departments of Biological Sciences and edgar.r.wood@mindspring.com.
authors.forename: Edgar R
authors.initials: ER
authors.lastname: Wood
authors.affiliation: From the Departments of Biological Sciences and.
authors.forename: Randy
authors.initials: R
authors.lastname: Bledsoe
authors.affiliation: ELT Boston, GlaxoSmithKline, Waltham, Massachusetts 02451.
authors.forename: Jing
authors.initials: J
authors.lastname: Chai
authors.affiliation: Antiviral Discovery Performance Unit, GlaxoSmithKline, Research Triangle Park, North Carolina 27709 and.
authors.forename: Philias
authors.initials: P
authors.lastname: Daka
authors.affiliation: ELT Boston, GlaxoSmithKline, Waltham, Massachusetts 02451.
authors.forename: Hongfeng
authors.initials: H
authors.lastname: Deng
authors.affiliation: ELT Boston, GlaxoSmithKline, Waltham, Massachusetts 02451.
authors.forename: Yun
authors.initials: Y
authors.lastname: Ding
authors.affiliation: From the Departments of Biological Sciences and.
authors.forename: Sarah
authors.initials: S
authors.lastname: Harris-Gurley
authors.affiliation: From the Departments of Biological Sciences and.
authors.forename: Luz Helena
authors.initials: LH
authors.lastname: Kryn
authors.affiliation: From the Departments of Biological Sciences and.
authors.forename: Eldridge
authors.initials: E
authors.lastname: Nartey
authors.affiliation: From the Departments of Biological Sciences and.
authors.forename: James
authors.initials: J
authors.lastname: Nichols
authors.affiliation: Chemical Sciences.
authors.forename: Robert T
authors.initials: RT
authors.lastname: Nolte
authors.affiliation: ELT Boston, GlaxoSmithKline, Waltham, Massachusetts 02451.
authors.forename: Ninad
authors.initials: N
authors.lastname: Prabhu
authors.affiliation: ELT Boston, GlaxoSmithKline, Waltham, Massachusetts 02451.
authors.forename: Cecil
authors.initials: C
authors.lastname: Rise
authors.affiliation: Antiviral Discovery Performance Unit, GlaxoSmithKline, Research Triangle Park, North Carolina 27709 and.
authors.forename: Timothy
authors.initials: T
authors.lastname: Sheahan
authors.affiliation: Antiviral Discovery Performance Unit, GlaxoSmithKline, Research Triangle Park, North Carolina 27709 and.
authors.forename: J Brad
authors.initials: JB
authors.lastname: Shotwell
authors.affiliation: From the Departments of Biological Sciences and.
authors.forename: Danielle
authors.initials: D
authors.lastname: Smith
authors.affiliation: Antiviral Discovery Performance Unit, GlaxoSmithKline, Research Triangle Park, North Carolina 27709 and.
authors.forename: Vince
authors.initials: V
authors.lastname: Tai
authors.affiliation: From the Departments of Biological Sciences and.
authors.forename: J David
authors.initials: JD
authors.lastname: Taylor
authors.affiliation: From the Departments of Biological Sciences and.
authors.forename: Ginger
authors.initials: G
authors.lastname: Tomberlin
authors.affiliation: Chemical Sciences.
authors.forename: Liping
authors.initials: L
authors.lastname: Wang
authors.affiliation: From the Departments of Biological Sciences and.
authors.forename: Bruce
authors.initials: B
authors.lastname: Wisely
authors.affiliation: Antiviral Discovery Performance Unit, GlaxoSmithKline, Research Triangle Park, North Carolina 27709 and.
authors.forename: Shihyun
authors.initials: S
authors.lastname: You
authors.affiliation: ELT Boston, GlaxoSmithKline, Waltham, Massachusetts 02451.
authors.forename: Bing
authors.initials: B
authors.lastname: Xia
authors.affiliation: Antiviral Discovery Performance Unit, GlaxoSmithKline, Research Triangle Park, North Carolina 27709 and.
authors.forename: Hamilton
authors.initials: H
authors.lastname: Dickson
mesh_terms: D015088:2',5'-Oligoadenylate Synthetase / Q000235:genetics / Q000276:immunology*; D000227:Adenine Nucleotides / Q000276:immunology / Q000378:metabolism; D000998:Antiviral Agents / Q000138:chemical synthesis / Q000494:pharmacology*; D018360:Crystallography, X-Ray; D004686:Encephalomyocarditis virus / Q000235:genetics / Q000378:metabolism; D004722:Endoribonucleases / Q000235:genetics / Q000276:immunology*; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005095:Exoribonucleases / Q000037:antagonists & inhibitors / Q000737:chemistry* / Q000235:genetics / Q000276:immunology; D005786:Gene Expression Regulation; D055786:Gene Knockout Techniques; D006367:HeLa Cells; D006801:Humans; D007113:Immunity, Innate; D016898:Interferon-alpha / Q000494:pharmacology; D008958:Models, Molecular; D009843:Oligoribonucleotides / Q000276:immunology / Q000378:metabolism; D011070:Poly I-C / Q000494:pharmacology; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D012136:Respiratory Syncytial Viruses / Q000235:genetics / Q000378:metabolism; D012229:Rhinovirus / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction; D054852:Small Molecule Libraries / Q000138:chemical synthesis / Q000494:pharmacology*; D013329:Structure-Activity Relationship
chemical_list: D000227:Adenine Nucleotides; D000998:Antiviral Agents; D016898:Interferon-alpha; D009843:Oligoribonucleotides; D011994:Recombinant Proteins; D054852:Small Molecule Libraries; C023505:2',5'-oligoadenylate; D015088:2',5'-Oligoadenylate Synthetase; D004722:Endoribonucleases; D005095:Exoribonucleases; C568404:PDE12 protein, human; C031956:2-5A-dependent ribonuclease; D011070:Poly I-C
keywords: antiviral agent; gene knockout; innate immunity; interferon; protein structure