title: Pneumocystis jirovecii Rtt109, a novel drug target for Pneumocystis pneumonia in immunosuppressed humans.
abstract: Pneumocystis pneumonia (PcP) is a significant cause of morbidity and mortality in immunocompromised patients. In humans, PcP is caused by the opportunistic fungal species Pneumocystis jirovecii. Progress in Pneumocystis research has been hampered by a lack of viable in vitro culture methods, which limits laboratory access to human-derived organisms for drug testing. Consequently, most basic drug discovery research for P. jirovecii is performed using related surrogate organisms such as Pneumocystis carinii, which is derived from immunosuppressed rodents. While these studies provide useful insights, important questions arise about interspecies variations and the relative utility of identified anti-Pneumocystis agents against human P. jirovecii. Our recent work has identified the histone acetyltransferase (HAT) Rtt109 in P. carinii (i.e., PcRtt109) as a potential therapeutic target for PcP, since Rtt109 HATs are widely conserved in fungi but are absent in humans. To further address the potential utility of this target in human disease, we now demonstrate the presence of a functional Rtt109 orthologue in the clinically relevant fungal pathogen P. jirovecii (i.e., PjRtt109). In a fashion similar to that of Pcrtt109, Pjrtt109 restores H3K56 acetylation and genotoxic resistance in rtt109-null yeast. Recombinant PjRtt109 is an active HAT in vitro, with activity comparable to that of PcRtt109 and yeast Rtt109. PjRtt109 HAT activity is also enhanced by the histone chaperone Asf1 in vitro. PjRtt109 and PcRtt109 showed similar low micromolar sensitivities to two reported small-molecule HAT inhibitors in vitro. Together, these results demonstrate that PjRtt109 is a functional Rtt109 HAT, and they support the development of anti-Pneumocystis agents directed at Rtt109-catalyzed histone acetylation as a novel therapeutic target for human PcP.
authors.affiliation: Department of Molecular Pharmacology and Experimental Therapeutics, Mayo Clinic College of Medicine, Rochester, Minnesota, USA Medical Scientist Training Program, Mayo Clinic College of Medicine, Rochester, Minnesota, USA.
authors.forename: Jayme L
authors.initials: JL
authors.lastname: Dahlin
authors.affiliation: Thoracic Diseases Research Unit, Mayo Clinic College of Medicine, Rochester, Minnesota, USA Department of Biochemistry and Molecular Biology, Mayo Clinic College of Medicine, Rochester, Minnesota, USA.
authors.forename: Theodore
authors.initials: T
authors.lastname: Kottom
authors.affiliation: Department of Biochemistry and Molecular Biology, Mayo Clinic College of Medicine, Rochester, Minnesota, USA.
authors.forename: Junhong
authors.initials: J
authors.lastname: Han
authors.affiliation: Department of Biochemistry and Molecular Biology, Mayo Clinic College of Medicine, Rochester, Minnesota, USA.
authors.forename: Hui
authors.initials: H
authors.lastname: Zhou
authors.affiliation: Institute for Therapeutics Discovery and Development, University of Minnesota, Minneapolis, Minnesota, USA.
authors.forename: Michael A
authors.initials: MA
authors.lastname: Walters
authors.affiliation: Department of Biochemistry and Molecular Biology, Mayo Clinic College of Medicine, Rochester, Minnesota, USA.
authors.forename: Zhiguo
authors.initials: Z
authors.lastname: Zhang
authors.affiliation: Thoracic Diseases Research Unit, Mayo Clinic College of Medicine, Rochester, Minnesota, USA Department of Biochemistry and Molecular Biology, Mayo Clinic College of Medicine, Rochester, Minnesota, USA Division of Pulmonary and Critical Care, Department of Internal Medicine, Mayo Clinic College of Medicine, Rochester, Minnesota, USA limper.andrew@mayo.edu.
authors.forename: Andrew H
authors.initials: AH
authors.lastname: Limper
mesh_terms: D017088:AIDS-Related Opportunistic Infections / Q000188:drug therapy* / Q000382:microbiology*; D000595:Amino Acid Sequence; D004271:DNA, Fungal / Q000235:genetics; D004305:Dose-Response Relationship, Drug; D025141:Drug Resistance, Fungal; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005656:Fungal Proteins / Q000187:drug effects* / Q000235:genetics*; D016681:Genome, Fungal / Q000235:genetics; D051548:Histone Acetyltransferases / Q000037:antagonists & inhibitors / Q000187:drug effects* / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D008969:Molecular Sequence Data; D045363:Pneumocystis carinii / Q000187:drug effects* / Q000235:genetics; D011020:Pneumonia, Pneumocystis / Q000188:drug therapy* / Q000382:microbiology*; D012441:Saccharomyces cerevisiae / Q000378:metabolism
chemical_list: D004271:DNA, Fungal; D004791:Enzyme Inhibitors; D005656:Fungal Proteins; D051548:Histone Acetyltransferases
references.citation: J Med Chem. 2010 Apr 8;53(7):2719-40
references.pmid: 20131845
references.citation: Am J Respir Cell Mol Biol. 2011 Dec;45(6):1142-6
references.pmid: 21642588
references.citation: Am J Respir Cell Mol Biol. 2010 Aug;43(2):192-200
references.pmid: 19783787
references.citation: Cell. 2008 Jul 25;134(2):244-55
references.pmid: 18662540
references.citation: Chest. 2005 Aug;128(2):573-9
references.pmid: 16100140
references.citation: J Protozool. 1991 Nov-Dec;38(6):126S-127S
references.pmid: 1818134
references.citation: Microbiology (Reading). 2004 May;150(Pt 5):1173-1178
references.pmid: 15133077
references.citation: Cell. 2008 Jul 25;134(2):231-43
references.pmid: 18662539
references.citation: Curr Drug Targets. 2012 Nov;13(12):1575-85
references.pmid: 22934582
references.citation: Bioorg Med Chem Lett. 2013 May 15;23(10):2853-9
references.pmid: 23587423
references.citation: J Clin Microbiol. 2008 Mar;46(3):966-71
references.pmid: 18216217
references.citation: Nature. 2005 Jul 14;436(7048):294-8
references.pmid: 16015338
references.citation: PLoS One. 2013 Nov 18;8(11):e78877
references.pmid: 24260132
references.citation: EMBO J. 2009 Jul 8;28(13):1878-89
references.pmid: 19407812
references.citation: J Clin Microbiol. 2013 Oct;51(10):3380-8
references.pmid: 23903553
references.citation: Nat Med. 2010 Jul;16(7):774-80
references.pmid: 20601951
references.citation: J Biol Chem. 2007 May 18;282(20):15040-7
references.pmid: 17369611
references.citation: Nature. 2009 May 7;459(7243):113-7
references.pmid: 19270680
references.citation: J Eukaryot Microbiol. 2006;53 Suppl 1:S149-50
references.pmid: 17169038
references.citation: Chest. 1996 Feb;109(2):586-7
references.pmid: 8620753
references.citation: Infect Immun. 2013 Jun;81(6):2268-75
references.pmid: 23569117
references.citation: Methods. 2005 Aug;36(4):321-31
references.pmid: 16085424
references.citation: J Mol Biol. 2000 Mar 31;297(3):645-57
references.pmid: 10731418
references.citation: Science. 2007 Feb 2;315(5812):653-5
references.pmid: 17272723
references.citation: Cancer Lett. 2005 Apr 18;221(1):29-39
references.pmid: 15797624
references.citation: Thorax. 2009 Dec;64(12):1070-6
references.pmid: 19825785
references.citation: J Cell Biochem. 2005 Sep 1;96(1):155-69
references.pmid: 16052481
references.citation: Free Radic Biol Med. 2007 Apr 15;42(8):1211-21
references.pmid: 17382202
references.citation: Antimicrob Agents Chemother. 2000 Mar;44(3):713-9
references.pmid: 10681344
references.citation: J Eukaryot Microbiol. 2006 May-Jun;53(3):157-64
references.pmid: 16677337
references.citation: Am J Respir Cell Mol Biol. 2011 Jun;44(6):768-76
references.pmid: 20656950
references.citation: Proc Am Thorac Soc. 2011 Jun;8(3):294-300
references.pmid: 21653531
references.citation: Proc Natl Acad Sci U S A. 2008 Aug 26;105(34):12236-41
references.pmid: 18719104
references.citation: Infect Immun. 2009 Mar;77(3):1053-60
references.pmid: 19124601
references.citation: Nutr Cancer. 2012;64(8):1211-1218
references.pmid: 23137051
references.citation: Biochem Pharmacol. 2009 May 1;77(9):1513-21
references.pmid: 19426689
references.citation: Int J Radiat Oncol Biol Phys. 2012 Nov 1;84(3):815-21
references.pmid: 22417805
references.citation: J Infect Chemother. 2012 Dec;18(6):793-806
references.pmid: 22864454
references.citation: Future Microbiol. 2010 Aug;5(8):1257-67
references.pmid: 20722602
references.citation: J Biol Chem. 2007 Sep 28;282(39):28587-28596
references.pmid: 17690098
references.citation: J Biol Chem. 2004 Jan 23;279(4):2817-24
references.pmid: 14576156
references.citation: Nat Rev Microbiol. 2007 Apr;5(4):298-308
references.pmid: 17363968
references.citation: Carcinogenesis. 2000 Jun;21(6):1183-9
references.pmid: 10837008
references.citation: J Infect Dis. 2009 Nov 15;200(10):1616-22
references.pmid: 19795979
references.citation: Food Funct. 2010 Dec;1(3):301-7
references.pmid: 21776480
references.citation: Future Microbiol. 2010 Jan;5(1):43-65
references.pmid: 20020829
references.citation: mBio. 2013 Apr 16;4(2):e00224
references.pmid: 23592264
references.citation: Proc Natl Acad Sci U S A. 2010 Jan 26;107(4):1594-9
references.pmid: 20080646
references.citation: Rheumatology (Oxford). 2011 Mar;50(3):569-77
references.pmid: 21097450
references.citation: Curr Opin Pulm Med. 2008 May;14(3):228-34
references.pmid: 18427246
references.citation: mBio. 2012 Dec 26;4(1):e00428-12
references.pmid: 23269827
references.citation: Science. 2007 Feb 2;315(5812):649-52
references.pmid: 17272722
references.citation: Crit Care. 2008;12(1):R28
references.pmid: 18304356
references.citation: Antimicrob Agents Chemother. 2006 Jul;50(7):2337-43
references.pmid: 16801410
references.citation: Immunol Rev. 2009 Jul;230(1):38-50
references.pmid: 19594628
references.citation: J Infect Dis. 1998 Dec;178(6):1767-75
references.pmid: 9815231
references.citation: Diagn Microbiol Infect Dis. 2011 Feb;69(2):145-52
references.pmid: 21251557
references.citation: Chest. 2000 Sep;118(3):704-11
references.pmid: 10988192
references.citation: Expert Rev Mol Med. 2005 Nov 14;7(26):1-16
references.pmid: 16842636
references.citation: J Biol Chem. 2004 Aug 6;279(32):33716-26
references.pmid: 15155757
references.citation: FASEB J. 2013 May;27(5):1962-72
references.pmid: 23392351
references.citation: Clin Microbiol Infect. 2011 Apr;17(4):595-602
references.pmid: 20673271
references.citation: Int J STD AIDS. 2009 Sep;20(9):652-3
references.pmid: 19710343
references.citation: Med Mycol. 2005 Feb;43(1):1-19
references.pmid: 15712604
references.citation: J Med Chem. 1998 Apr 9;41(8):1263-71
references.pmid: 9548816
references.citation: J Agric Food Chem. 2011 Feb 9;59(3):1031-40
references.pmid: 21214247
references.citation: J Biol Chem. 2002 Mar 8;277(10):8749-54
references.pmid: 11773058
references.citation: Ther Adv Respir Dis. 2011 Feb;5(1):41-59
references.pmid: 20736243
references.citation: J Med Chem. 2009 Jan 22;52(2):267-77
references.pmid: 19086895
references.citation: Exp Parasitol. 1990 Jan;70(1):100-6
references.pmid: 2295322
references.citation: Antimicrob Agents Chemother. 2013 Jun;57(6):2669-77
references.pmid: 23545530
references.citation: Biochemistry. 2009 Mar 3;48(8):1702-11
references.pmid: 19196009
references.citation: J Eukaryot Microbiol. 1993 Mar-Apr;40(2):188-95
references.pmid: 8461892
references.citation: Infect Immun. 1996 Nov;64(11):4643-7
references.pmid: 8890219
references.citation: Mol Cell. 2007 Mar 9;25(5):703-12
references.pmid: 17320445
references.citation: J Biol Chem. 2007 Jan 12;282(2):1334-40
references.pmid: 17107956
references.citation: Mol Cancer Ther. 2012 Oct;11(10):2193-201
references.pmid: 22821148
references.citation: Emerg Infect Dis. 2004 Oct;10(10):1721-8
references.pmid: 15504256
references.citation: J Biol Chem. 2011 Jul 15;286(28):24694-701
references.pmid: 21606491
references.citation: J Med Microbiol. 2013 Aug;62(Pt 8):1170-1174
references.pmid: 23699069
references.citation: Annu Rev Microbiol. 2010;64:431-52
references.pmid: 20528694