title: Proteolytic elimination of N-myristoyl modifications by the Shigella virulence factor IpaJ.
abstract: Protein N-myristoylation is a 14-carbon fatty-acid modification that is conserved across eukaryotic species and occurs on nearly 1% of the cellular proteome. The ability of the myristoyl group to facilitate dynamic protein-protein and protein-membrane interactions (known as the myristoyl switch) makes it an essential feature of many signal transduction systems. Thus pathogenic strategies that facilitate protein demyristoylation would markedly alter the signalling landscape of infected host cells. Here we describe an irreversible mechanism of protein demyristoylation catalysed by invasion plasmid antigen J (IpaJ), a previously uncharacterized Shigella flexneri type III effector protein with cysteine protease activity. A yeast genetic screen for IpaJ substrates identified ADP-ribosylation factor (ARF)1p and ARF2p, small molecular mass GTPases that regulate cargo transport through the Golgi apparatus. Mass spectrometry showed that IpaJ cleaved the peptide bond between N-myristoylated glycine-2 and asparagine-3 of human ARF1, thereby providing a new mechanism for host secretory inhibition by a bacterial pathogen. We further demonstrate that IpaJ cleaves an array of N-myristoylated proteins involved in cellular growth, signal transduction, autophagasome maturation and organelle function. Taken together, these findings show a previously unrecognized pathogenic mechanism for the site-specific elimination of N-myristoyl protein modification.
authors.affiliation: Department of Microbiology, University of Texas Southwestern Medical Center, 5323 Harry Hines Boulevard, Dallas, Texas 75390-8816, USA.
authors.forename: Nikolay
authors.initials: N
authors.lastname: Burnaevskiy
authors.forename: Thomas G
authors.initials: TG
authors.lastname: Fox
authors.forename: Daniel A
authors.initials: DA
authors.lastname: Plymire
authors.forename: James M
authors.initials: JM
authors.lastname: Ertelt
authors.forename: Bethany A
authors.initials: BA
authors.lastname: Weigele
authors.forename: Andrey S
authors.initials: AS
authors.lastname: Selyunin
authors.forename: Sing Sing
authors.initials: SS
authors.lastname: Way
authors.forename: Steven M
authors.initials: SM
authors.lastname: Patrie
authors.forename: Neal M
authors.initials: NM
authors.lastname: Alto
mesh_terms: D020823:ADP-Ribosylation Factor 1 / Q000737:chemistry / Q000378:metabolism; D020727:ADP-Ribosylation Factors / Q000378:metabolism; D000595:Amino Acid Sequence; D000818:Animals; D000942:Antigens, Bacterial / Q000378:metabolism*; D001216:Asparagine / Q000378:metabolism; D001343:Autophagy; D055162:Biocatalysis; D057056:Cysteine Proteases / Q000378:metabolism; D004405:Dysentery, Bacillary; D005260:Female; D005998:Glycine / Q000378:metabolism; D006056:Golgi Apparatus / Q000378:metabolism / Q000473:pathology; D057809:HEK293 Cells; D006367:HeLa Cells; D006801:Humans; D008089:Listeria monocytogenes / Q000502:physiology; D051379:Mice; D008810:Mice, Inbred C57BL; D008969:Molecular Sequence Data; D019814:Myristic Acid / Q000378:metabolism*; D010588:Phagosomes / Q000378:metabolism; D011499:Protein Processing, Post-Translational; D059748:Proteolysis; D012441:Saccharomyces cerevisiae; D029701:Saccharomyces cerevisiae Proteins / Q000378:metabolism; D016415:Sequence Alignment; D012763:Shigella flexneri / Q000201:enzymology / Q000378:metabolism*; D015398:Signal Transduction; D013379:Substrate Specificity; D014774:Virulence; D037521:Virulence Factors / Q000378:metabolism*
chemical_list: D000942:Antigens, Bacterial; C110952:IpaJ protein, Shigella flexneri; D029701:Saccharomyces cerevisiae Proteins; C533229:VirA protein, Shigella flexneri; D037521:Virulence Factors; D019814:Myristic Acid; D001216:Asparagine; D057056:Cysteine Proteases; C116686:ARF2 protein, S cerevisiae; D020823:ADP-Ribosylation Factor 1; D020727:ADP-Ribosylation Factors; D005998:Glycine
references.citation: J Am Chem Soc. 2009 Apr 8;131(13):4967-75
references.pmid: 19281244
references.citation: Anal Chem. 2011 Dec 15;83(24):9586-92
references.pmid: 22017354
references.citation: Methods Enzymol. 2008;439:131-43
references.pmid: 18374161
references.citation: Cell. 1991 Jan 25;64(2):425-37
references.pmid: 1988155
references.citation: Science. 2000 Feb 4;287(5454):826-30
references.pmid: 10657290
references.citation: Proc Natl Acad Sci U S A. 2000 Jun 6;97(12):6640-5
references.pmid: 10829079
references.citation: Infect Immun. 1998 Apr;66(4):1342-8
references.pmid: 9529051
references.citation: Curr Opin Immunol. 2011 Aug;23(4):448-55
references.pmid: 21763117
references.citation: Nucleic Acids Res. 2008 Apr;36(7):2295-300
references.pmid: 18287115
references.citation: Crit Rev Immunol. 2003;23(5-6):371-401
references.pmid: 15030306
references.citation: Proc Natl Acad Sci U S A. 1989 Jun;86(11):4012-6
references.pmid: 2726763
references.citation: Proc Natl Acad Sci U S A. 2008 Dec 9;105(49):19211-6
references.pmid: 19050071
references.citation: Nucleic Acids Res. 2005 Jul 1;33(Web Server issue):W244-8
references.pmid: 15980461
references.citation: J Biol Chem. 2001 Oct 26;276(43):39501-4
references.pmid: 11527981
references.citation: Structure. 2009 Jan 14;17(1):79-87
references.pmid: 19141284
references.citation: Proc Natl Acad Sci U S A. 2002 Feb 19;99(4):1774-9
references.pmid: 11842225
references.citation: Nature. 2011 Jan 6;469(7328):107-11
references.pmid: 21170023
references.citation: Cell Host Microbe. 2012 Sep 13;12(3):381-9
references.pmid: 22980334
references.citation: Trends Microbiol. 2001 Apr;9(4):164-8
references.pmid: 11286880
references.citation: Immunity. 1999 Jan;10(1):29-38
references.pmid: 10023768
references.citation: PLoS Pathog. 2008 Jan;4(1):e9
references.pmid: 18208325
references.citation: Genome Biol. 2004;5(3):R21
references.pmid: 15003124
references.citation: Microb Pathog. 1997 Dec;23(6):357-69
references.pmid: 9441862
references.citation: Anal Chem. 2004 Jun 1;76(11):197A-203A
references.pmid: 15190879
references.citation: Nat Struct Mol Biol. 2010 Jul;17(7):876-81
references.pmid: 20601958
references.citation: Cell Microbiol. 1999 Jul;1(1):33-49
references.pmid: 11207539
references.citation: FEBS Lett. 2009 Dec 3;583(23):3872-9
references.pmid: 19879269
references.citation: J Cell Biol. 1997 Jun 16;137(6):1211-28
references.pmid: 9182657
references.citation: Cell. 2012 Aug 31;150(5):1029-41
references.pmid: 22939626
references.citation: J Biol Chem. 1996 Jan 19;271(3):1573-8
references.pmid: 8576155
references.citation: Proc Natl Acad Sci U S A. 1985 Jul;82(14):4625-8
references.pmid: 2991884
references.citation: Science. 2012 Nov 23;338(6110):1072-6
references.pmid: 23112293
references.citation: Cell. 1997 Oct 17;91(2):253-62
references.pmid: 9346242
references.citation: Plasmid. 2002 Jul;48(1):59-63
references.pmid: 12206756
references.citation: Cell. 2002 May 31;109(5):575-88
references.pmid: 12062101
references.citation: Trends Biochem Sci. 1995 Jul;20(7):272-6
references.pmid: 7667880