title: Structural basis for inhibition of the deadenylase activity of human CNOT6L.
abstract: Human CNOT6L/CCR4, a member of the endonuclease-exonuclease-phosphatase (EEP) family enzymes, is one of the two deadenylase enzymes in the conserved CCR4-NOT complex. Here, we report inhibitor-bound crystal structures of the human CNOT6L nuclease domain in complex with the nucleotide CMP and the aminoglycoside neomycin. Deadenylase activity assays show that nucleotides are effective inhibitors of both CNOT6L and CNOT7, with AMP more effective than other nucleotides, and that neomycin is a weak deadenylase inhibitor. Structural analysis shows that all inhibitors occupy the substrate and magnesium-binding sites of CNOT6L, suggesting that inhibitors compete with both substrate and divalent magnesium ions for overlapping binding sites.
authors.affiliation: College of Life Sciences, Nankai University, Tianjin, China.
authors.forename: Qionglin
authors.initials: Q
authors.lastname: Zhang
authors.affiliation: College of Life Sciences, Nankai University, Tianjin, China.|State Key Laboratory of Medicinal Chemical Biology, Nankai University, Tianjin, China.
authors.forename: Dongke
authors.initials: D
authors.lastname: Yan
authors.affiliation: College of Life Sciences, Nankai University, Tianjin, China.|State Key Laboratory of Medicinal Chemical Biology, Nankai University, Tianjin, China.
authors.forename: Erhong
authors.initials: E
authors.lastname: Guo
authors.affiliation: College of Life Sciences, Nankai University, Tianjin, China.
authors.forename: Bojian
authors.initials: B
authors.lastname: Ding
authors.affiliation: College of Life Sciences, Nankai University, Tianjin, China.
authors.forename: Wen
authors.initials: W
authors.lastname: Yang
authors.affiliation: College of Life Sciences, Nankai University, Tianjin, China.
authors.forename: Ruihua
authors.initials: R
authors.lastname: Liu
authors.affiliation: Cell Signal Unit, Okinawa Institute of Science and Technology Graduate University, Kunigami, Okinawa, Japan.
authors.forename: Tadashi
authors.initials: T
authors.lastname: Yamamoto
authors.affiliation: College of Life Sciences, Nankai University, Tianjin, China.|State Key Laboratory of Medicinal Chemical Biology, Nankai University, Tianjin, China.
authors.forename: Mark
authors.identifier: 0000-0001-5173-5080
authors.initials: M
authors.lastname: Bartlam
mesh_terms: D001665:Binding Sites; D018360:Crystallography, X-Ray; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005095:Exoribonucleases; D006801:Humans; D020128:Inhibitory Concentration 50; D008274:Magnesium / Q000378:metabolism; D009355:Neomycin / Q000737:chemistry / Q000378:metabolism; D009711:Nucleotides / Q000378:metabolism; D012097:Repressor Proteins; D012260:Ribonucleases / Q000037:antagonists & inhibitors* / Q000737:chemistry*; D013329:Structure-Activity Relationship; D014157:Transcription Factors / Q000037:antagonists & inhibitors / Q000378:metabolism
chemical_list: D004791:Enzyme Inhibitors; D009711:Nucleotides; D012097:Repressor Proteins; D014157:Transcription Factors; C521732:CNOT6L protein, human; C545829:CNOT7 protein, human; D005095:Exoribonucleases; D012260:Ribonucleases; C445084:mRNA deadenylase; D009355:Neomycin; D008274:Magnesium
keywords: CCR4-NOT complex; crystal structure; deadenylase; inhibitor