title: Methylthioadenosine (MTA) Regulates Liver Cells Proteome and Methylproteome: Implications in Liver Biology and Disease.
abstract: Methylthioadenosine phosphorylase (MTAP), a key enzyme in the adenine and methionine salvage pathways, catalyzes the hydrolysis of methylthioadenosine (MTA), a compound suggested to affect pivotal cellular processes in part through the regulation of protein methylation. MTAP is expressed in a wide range of cell types and tissues, and its deletion is common to cancer cells and in liver injury. The aim of this study was to investigate the proteome and methyl proteome alterations triggered by MTAP deficiency in liver cells to define novel regulatory mechanisms that may explain the pathogenic processes of liver diseases. iTRAQ analysis resulted in the identification of 216 differential proteins (p < 0.05) that suggest deregulation of cellular pathways as those mediated by ERK or NFκB. R-methyl proteome analysis led to the identification of 74 differentially methylated proteins between SK-Hep1 and SK-Hep1+ cells, including 116 new methylation sites. Restoring normal MTA levels in SK-Hep1+ cells parallels the specific methylation of 56 proteins, including KRT8, TGF, and CTF8A, which provides a novel regulatory mechanism of their activity with potential implications in carcinogenesis. Inhibition of RNA-binding proteins methylation is especially relevant upon accumulation of MTA. As an example, methylation of quaking protein in Arg(242) and Arg(256) in SK-Hep1+ cells may play a pivotal role in the regulation of its activity as indicated by the up-regulation of its target protein p27(kip1) The phenotype associated with a MTAP deficiency was further verified in the liver of MTAP± mice. Our data support that MTAP deficiency leads to MTA accumulation and deregulation of central cellular pathways, increasing proliferation and decreasing the susceptibility to chemotherapeutic drugs, which involves differential protein methylation. Data are available via ProteomeXchange with identifier PXD002957 (http://www.ebi.ac.uk/pride/archive/projects/PXD002957).
authors.affiliation: From the §Department of Hepatology, Proteomics Laboratory, CIMA, University of Navarra; CIBERehd; IDISNA, Pamplona, 31008 Spain.
authors.forename: Emilie
authors.initials: E
authors.lastname: Bigaud
authors.affiliation: From the §Department of Hepatology, Proteomics Laboratory, CIMA, University of Navarra; CIBERehd; IDISNA, Pamplona, 31008 Spain fjcorrales@unav.es.
authors.forename: Fernando J
authors.initials: FJ
authors.lastname: Corrales
mesh_terms: D000818:Animals; D049109:Cell Proliferation; D002478:Cells, Cultured; D056486:Chemical and Drug Induced Liver Injury / Q000201:enzymology / Q000235:genetics / Q000473:pathology*; D003839:Deoxyadenosines / Q000378:metabolism*; D004195:Disease Models, Animal; D005260:Female; D005466:Fluorocarbons / Q000009:adverse effects; D005786:Gene Expression Regulation; D056945:Hep G2 Cells; D022781:Hepatocytes / Q000166:cytology* / Q000378:metabolism / Q000473:pathology; D006801:Humans; D008297:Male; D008745:Methylation; D051379:Mice; D020543:Proteome / Q000378:metabolism*; D011683:Purine-Nucleoside Phosphorylase / Q000172:deficiency* / Q000235:genetics*; D015398:Signal Transduction; D013872:Thionucleosides / Q000378:metabolism*
chemical_list: D003839:Deoxyadenosines; D005466:Fluorocarbons; D020543:Proteome; D013872:Thionucleosides; C008500:5'-methylthioadenosine; C035066:carbon tetrafluoride; D011683:Purine-Nucleoside Phosphorylase; C021784:5'-methylthioadenosine phosphorylase
references.citation: Cancer Biol Ther. 2011 Apr 1;11(7):627-32
references.pmid: 21301207
references.citation: Am J Pathol. 2011 Mar;178(3):1145-52
references.pmid: 21356366
references.citation: J Biol Chem. 2003 Dec 12;278(50):49868-73
references.pmid: 14506228
references.citation: J Biol Chem. 2010 Apr 23;285(17):12695-705
references.pmid: 20159986
references.citation: Rev Physiol Biochem Pharmacol. 1994;125:1-42
references.pmid: 7984872
references.citation: J Cell Sci. 2009 Mar 1;122(Pt 5):667-77
references.pmid: 19208762
references.citation: Hepatology. 2002 Feb;35(2):274-80
references.pmid: 11826399
references.citation: Int J Biochem. 1979;10(11):901-4
references.pmid: 229009
references.citation: Neuron. 2014 Mar 5;81(5):1024-39
references.pmid: 24607226
references.citation: Mol Cell. 2005 Apr 29;18(3):263-72
references.pmid: 15866169
references.citation: J Cell Physiol. 2006 Jul;208(1):154-60
references.pmid: 16547966
references.citation: Hepatology. 2012 Apr;55(4):1237-48
references.pmid: 22095636
references.citation: Mol Cell Proteomics. 2014 Jan;13(1):372-87
references.pmid: 24129315
references.citation: Gastroenterology. 2009 Mar;136(3):1025-36
references.pmid: 18983843
references.citation: Cancer Res. 2009 Jul 15;69(14):5961-9
references.pmid: 19567676
references.citation: Proc Natl Acad Sci U S A. 1994 Jan 18;91(2):674-8
references.pmid: 7904757
references.citation: J Proteome Res. 2008 Mar;7(3):1001-6
references.pmid: 18220335
references.citation: Nat Neurosci. 2005 Jan;8(1):27-33
references.pmid: 15568022
references.citation: Biochem Pharmacol. 1982 Feb 1;31(3):277-88
references.pmid: 6803807
references.citation: Cancer. 2009 Dec 15;115(24):5651-61
references.pmid: 19834957
references.citation: Gastroenterology. 1998 Feb;114(2):364-71
references.pmid: 9453498
references.citation: Biochem Pharmacol. 1981 Feb 1;30(3):189-99
references.pmid: 6164373
references.citation: Anal Chem. 1996 Mar 1;68(5):850-8
references.pmid: 8779443
references.citation: J Environ Sci Health C Environ Carcinog Ecotoxicol Rev. 2007 Jul-Sep;25(3):185-209
references.pmid: 17763046
references.citation: Int J Biochem Cell Biol. 2004 Nov;36(11):2125-30
references.pmid: 15313459
references.citation: J Biol Chem. 2012 Jun 29;287(27):22854-64
references.pmid: 22582387
references.citation: Cancer Res. 1988 Feb 15;48(4):759-74
references.pmid: 3123052
references.citation: J Biol Chem. 2013 Sep 27;288(39):27752-63
references.pmid: 23921388
references.citation: Mol Cell Biol. 2008 Dec;28(23):7066-80
references.pmid: 18809583
references.citation: FASEB J. 2000 Dec;14(15):2511-8
references.pmid: 11099469
references.citation: Hepatology. 2007 May;45(5):1306-12
references.pmid: 17464973
references.citation: Proteomics. 2014 Mar;14(6):680-4
references.pmid: 24449435
references.citation: J Biol Chem. 2007 Aug 10;282(32):23553-60
references.pmid: 17575274
references.citation: Cancer Res. 2006 Sep 1;66(17):8788-95
references.pmid: 16951195
references.citation: Biochem Pharmacol. 1991 Jun 15;41(12):2023-30
references.pmid: 1903946
references.citation: Carcinogenesis. 2011 Oct;32(10):1419-26
references.pmid: 21642356
references.citation: Biochim Biophys Acta. 2004 Nov 5;1690(3):276-84
references.pmid: 15511635
references.citation: Bioessays. 1999 Aug;21(8):704-9
references.pmid: 10440867
references.citation: Open Biol. 2015 Apr;5(4):150018
references.pmid: 25833379
references.citation: Hepatology. 2013 Oct;58(4):1296-305
references.pmid: 23505042
references.citation: Crit Rev Toxicol. 2003;33(2):105-36
references.pmid: 12708612
references.citation: Bull Exp Biol Med. 2012 Oct;153(6):878-81
references.pmid: 23113308
references.citation: Science. 2013 Jan 11;339(6116):222-6
references.pmid: 23118012
references.citation: Gastroenterology. 2010 Jan;138(1):231-40.e1-5
references.pmid: 19686745
references.citation: Stat Appl Genet Mol Biol. 2004;3:Article3
references.pmid: 16646809
references.citation: Nat Methods. 2013 Apr;10(4):339-42
references.pmid: 23455924
references.citation: Prog Nucleic Acid Res Mol Biol. 1998;61:65-131
references.pmid: 9752719
references.citation: Oncogene. 2006 May 18;25(21):3049-58
references.pmid: 16449978
references.citation: Nat Chem Biol. 2013 May;9(5):300-6
references.pmid: 23455543
references.citation: PLoS One. 2013;8(12):e80703
references.pmid: 24324622
references.citation: J Biol Chem. 2002 Nov 15;277(46):44028-34
references.pmid: 12215427
references.citation: Toxicol Sci. 2010 May;115(1):131-9
references.pmid: 20118189
references.citation: Pancreatology. 2015 Jul;15(4 Suppl):S49-54
references.pmid: 25841271
references.citation: Nat Rev Cancer. 2013 Aug;13(8):572-83
references.pmid: 23822983
references.citation: Nat Rev Cancer. 2010 Apr;10(4):301-9
references.pmid: 20332779
references.citation: Curr Opin Gastroenterol. 2012 May;28(3):209-16
references.pmid: 22450891
references.citation: Nat Biotechnol. 2014 Mar;32(3):223-6
references.pmid: 24727771
references.citation: FASEB J. 2010 Nov;24(11):4153-66
references.pmid: 20667976
references.citation: Toxicol Appl Pharmacol. 2010 Jan 1;242(1):47-55
references.pmid: 19796649
references.citation: Mol Biol Cell. 2003 Jan;14(1):274-87
references.pmid: 12529443
references.citation: Mol Cell Biol. 2004 Aug;24(16):7072-81
references.pmid: 15282307
references.citation: Br J Cancer. 2007 Aug 6;97(3):368-77
references.pmid: 17667909
references.citation: Eur J Biochem. 1979 Nov;101(2):317-24
references.pmid: 118001
references.citation: Science. 2009 May 22;324(5930):1029-33
references.pmid: 19460998
references.citation: Mol Cell. 2007 Jan 12;25(1):71-83
references.pmid: 17218272
references.citation: J Biol Chem. 2002 Mar 15;277(11):9062-8
references.pmid: 11782484
references.citation: Mol Cell Proteomics. 2011 Jan;10(1):M110.003335
references.pmid: 20807836
references.citation: J Biol Chem. 1969 Feb 25;244(4):682-93
references.pmid: 4889860
references.citation: Hepatology. 2006 Sep;44(3):521-6
references.pmid: 16941687
references.citation: Sci Signal. 2011 Sep 13;4(190):ra58
references.pmid: 21917714
references.citation: Eur J Cancer. 2013 Apr;49(6):1305-13
references.pmid: 23265702
references.citation: Nat Rev Mol Cell Biol. 2011 Oct 12;12(11):689
references.pmid: 21993294
references.citation: Oncol Rep. 1998 Nov-Dec;5(6):1385-8
references.pmid: 9769373
references.citation: Mol Cell Biol. 1995 May;15(5):2800-8
references.pmid: 7739561
references.citation: Ann Hepatol. 2013 Mar-Apr;12(2):183-9
references.pmid: 23396728
references.citation: Int J Biol Sci. 2011;7(5):536-50
references.pmid: 21552420
references.citation: Mol Med. 2002 Oct;8(10):655-63
references.pmid: 12477976
references.citation: J Biol Chem. 1990 Apr 15;265(11):6241-6
references.pmid: 2108157
references.citation: Mol Cell Proteomics. 2011 Jun;10(6):M111.009126
references.pmid: 21467216
references.citation: Biochem J. 2008 Apr 15;411(2):457-65
references.pmid: 18237276
references.citation: Clin Transl Oncol. 2014 Jun;16(6):561-6
references.pmid: 24105047
references.citation: Nat Commun. 2012;3:1072
references.pmid: 22990868
references.citation: Cell Mol Biol (Noisy-le-grand). 2009;55(1):29-37
references.pmid: 19267999
references.citation: Alcohol. 2002 Jul;27(3):193-8
references.pmid: 12163149
references.citation: J Gastroenterol Hepatol. 2008 Mar;23 Suppl 1:S73-7
references.pmid: 18336669