title: CyclinA2-Cyclin-dependent Kinase Regulates SAMHD1 Protein Phosphohydrolase Domain.
abstract: SAMHD1 is a nuclear deoxyribonucleoside triphosphate triphosphohydrolase that contributes to the control of cellular deoxyribonucleoside triphosphate (dNTP) pool sizes through dNTP hydrolysis and modulates the innate immune response to viruses. CyclinA2-CDK1/2 phosphorylates SAMHD1 at Thr-592, but how this modification controls SAMHD1 functions in proliferating cells is not known. Here, we show that SAMHD1 levels remain relatively unchanged during the cell division cycle in primary human T lymphocytes and in monocytic cell lines. Inactivation of the bipartite cyclinA2-CDK-binding site in the SAMHD1 C terminus described herein abolished SAMHD1 phosphorylation on Thr-592 during S and G2 phases thus interfering with DNA replication and progression of cells through S phase. The effects exerted by Thr-592 phosphorylation-defective SAMHD1 mutants were associated with activation of DNA damage checkpoint and depletion of dNTP concentrations to levels lower than those seen upon expression of wild type SAMHD1 protein. These disruptive effects were relieved by either mutation of the catalytic residues of the SAMHD1 phosphohydrolase domain or by a Thr-592 phosphomimetic mutation, thus linking the Thr-592 phosphorylation state to the control of SAMHD1 dNTPase activity. Our findings support a model in which phosphorylation of Thr-592 by cyclinA2-CDK down-modulates, but does not inactivate, SAMHD1 dNTPase in S phase, thereby fine-tuning SAMHD1 control of dNTP levels during DNA replication.
authors.affiliation: From the Department of Molecular Biology and Microbiology, Case Western Reserve School of Medicine, Cleveland, Ohio 44106.
authors.forename: Junpeng
authors.initials: J
authors.lastname: Yan
authors.affiliation: From the Department of Molecular Biology and Microbiology, Case Western Reserve School of Medicine, Cleveland, Ohio 44106.
authors.forename: Caili
authors.initials: C
authors.lastname: Hao
authors.affiliation: the Department of Structural Biology and.
authors.forename: Maria
authors.initials: M
authors.lastname: DeLucia
authors.affiliation: the Stowers Institute for Medical Research, Kansas City, Missouri 64110, and.
authors.forename: Selene
authors.initials: S
authors.lastname: Swanson
authors.affiliation: the Stowers Institute for Medical Research, Kansas City, Missouri 64110, and.
authors.forename: Laurence
authors.initials: L
authors.lastname: Florens
authors.affiliation: the Stowers Institute for Medical Research, Kansas City, Missouri 64110, and the Department of Pathology and Laboratory Medicine, University of Kansas Medical Center, Kansas City, Kansas 66160.
authors.forename: Michael P
authors.initials: MP
authors.lastname: Washburn
authors.affiliation: the Department of Structural Biology and Pittsburgh Center for HIV Protein Interactions, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania 15260.
authors.forename: Jinwoo
authors.initials: J
authors.lastname: Ahn
authors.affiliation: From the Department of Molecular Biology and Microbiology, Case Western Reserve School of Medicine, Cleveland, Ohio 44106, Pittsburgh Center for HIV Protein Interactions, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania 15260, jacek.skowronski@case.edu.
authors.forename: Jacek
authors.initials: J
authors.lastname: Skowronski
mesh_terms: D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D015153:Blotting, Western; D016203:CDC2 Protein Kinase; D002453:Cell Cycle / Q000502:physiology*; D002478:Cells, Cultured; D056751:Cyclin A2 / Q000235:genetics / Q000378:metabolism*; D051357:Cyclin-Dependent Kinase 2 / Q000235:genetics / Q000378:metabolism*; D018844:Cyclin-Dependent Kinases / Q000235:genetics / Q000378:metabolism*; D057809:HEK293 Cells; D006801:Humans; D006868:Hydrolysis; D007124:Immunoenzyme Techniques; D047468:Immunoprecipitation; D008969:Molecular Sequence Data; D009000:Monocytes / Q000166:cytology / Q000378:metabolism*; D020559:Monomeric GTP-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D009154:Mutation / Q000235:genetics; D010766:Phosphorylation; D017434:Protein Structure, Tertiary; D000076106:SAM Domain and HD Domain-Containing Protein 1; D017386:Sequence Homology, Amino Acid; D013601:T-Lymphocytes / Q000166:cytology / Q000378:metabolism*
chemical_list: C503721:CCNA2 protein, human; D056751:Cyclin A2; D016203:CDC2 Protein Kinase; C528899:CDK1 protein, human; C495894:CDK2 protein, human; D051357:Cyclin-Dependent Kinase 2; D018844:Cyclin-Dependent Kinases; D000076106:SAM Domain and HD Domain-Containing Protein 1; C484280:SAMHD1 protein, human; D020559:Monomeric GTP-Binding Proteins
keywords: Aicardi Goutieres Syndrome; DNA replication; SAM domain and HD domain-containing protein 1 (SAMHD1); cell cycle; dNTP pools; human immunodeficiency virus (HIV); innate immunity; restriction factor
references.citation: J Biol Chem. 2001 Jan 19;276(3):1993-7
references.pmid: 11067844
references.citation: J Biol Chem. 2014 Nov 21;289(47):32617-27
references.pmid: 25288794
references.citation: Retrovirology. 2012;9:87
references.pmid: 23092122
references.citation: Nature. 1970 Aug 15;227(5259):680-5
references.pmid: 5432063
references.citation: Mol Cell Biol. 1996 Dec;16(12):6623-33
references.pmid: 8943316
references.citation: Trends Biochem Sci. 1998 Dec;23(12):469-72
references.pmid: 9868367
references.citation: J Biol Chem. 2004 Dec 3;279(49):51545-53
references.pmid: 15452123
references.citation: J Biol Chem. 2006 Mar 24;281(12):7834-41
references.pmid: 16436374
references.citation: Annu Rev Biochem. 2006;75:681-706
references.pmid: 16756507
references.citation: Retrovirology. 2007;4:2
references.pmid: 17212817
references.citation: Proc Natl Acad Sci U S A. 2007 Jul 10;104(28):11778-83
references.pmid: 17609381
references.citation: Bioinformatics. 2007 Nov 1;23(21):2947-8
references.pmid: 17846036
references.citation: PLoS Pathog. 2008 May;4(5):e1000059
references.pmid: 18464893
references.citation: Mol Cell Proteomics. 2008 Sep;7(9):1598-608
references.pmid: 18463090
references.citation: Bioinformatics. 2009 May 1;25(9):1189-91
references.pmid: 19151095
references.citation: Genes Dev. 2010 Feb 15;24(4):333-8
references.pmid: 20159953
references.citation: Anal Chem. 2010 Mar 15;82(6):2272-81
references.pmid: 20166708
references.citation: Nature. 2011 Jun 30;474(7353):654-7
references.pmid: 21613998
references.citation: Nature. 2011 Jun 30;474(7353):658-61
references.pmid: 21720370
references.citation: Nature. 2011 Dec 15;480(7377):379-82
references.pmid: 22056990
references.citation: PLoS Pathog. 2011 Dec;7(12):e1002425
references.pmid: 22174685
references.citation: Crit Rev Biochem Mol Biol. 2012 Jan-Feb;47(1):50-63
references.pmid: 22050358
references.citation: J Biol Chem. 2011 Dec 23;286(51):43596-600
references.pmid: 22069334
references.citation: Cell Host Microbe. 2012 Feb 16;11(2):194-204
references.pmid: 22284954
references.citation: Cell Host Microbe. 2012 Feb 16;11(2):205-17
references.pmid: 22305291
references.citation: Nat Immunol. 2012 Mar;13(3):223-8
references.pmid: 22327569
references.citation: J Biol Chem. 2012 Apr 6;287(15):12550-8
references.pmid: 22362772
references.citation: J Biol Chem. 2012 Jun 22;287(26):21570-4
references.pmid: 22589553
references.citation: Nat Med. 2012 Nov;18(11):1682-7
references.pmid: 22972397
references.citation: Retrovirology. 2013;10:26
references.pmid: 23497255
references.citation: J Biol Chem. 2013 Mar 22;288(12):8101-10
references.pmid: 23364794
references.citation: J Biol Chem. 2013 Apr 12;288(15):10406-17
references.pmid: 23426366
references.citation: Cell Host Microbe. 2013 Apr 17;13(4):441-51
references.pmid: 23601106
references.citation: Cell Rep. 2013 Apr 25;3(4):1036-43
references.pmid: 23602554
references.citation: Handb Clin Neurol. 2013;113:1629-35
references.pmid: 23622384
references.citation: PLoS Pathog. 2013;9(6):e1003481
references.pmid: 23825958
references.citation: Proc Natl Acad Sci U S A. 2013 Aug 27;110(35):14272-7
references.pmid: 23858451
references.citation: Proc Natl Acad Sci U S A. 2013 Aug 27;110(35):14120-1
references.pmid: 23946423
references.citation: Cell Rep. 2013 Aug 29;4(4):689-96
references.pmid: 23972988
references.citation: J Biol Chem. 2013 Aug 30;288(35):25001-6
references.pmid: 23880768
references.citation: EMBO J. 2013 Sep 11;32(18):2454-62
references.pmid: 23872947
references.citation: J Virol. 2013 Nov;87(21):11516-24
references.pmid: 23966382
references.citation: PLoS Biol. 2004 Jan;2(1):E6
references.pmid: 14737186
references.citation: Nat Struct Mol Biol. 2013 Nov;20(11):1304-9
references.pmid: 24141705
references.citation: Nat Commun. 2013;4:2722
references.pmid: 24217394
references.citation: Proc Natl Acad Sci U S A. 2014 May 6;111(18):E1843-51
references.pmid: 24753578
references.citation: J Immunol. 2014 Aug 15;193(4):1988-97
references.pmid: 25015816
references.citation: Nat Med. 2014 Aug;20(8):936-41
references.pmid: 25038827
references.citation: Proc Natl Acad Sci U S A. 2014 Oct 14;111(41):E4305-14
references.pmid: 25267621
references.citation: Ann Rheum Dis. 2015 Mar;74(3):e17
references.pmid: 24445253