#PAGE_PARAMS# #ADS_HEAD_SCRIPTS# #MICRODATA#

CPAF: A Chlamydial Protease in Search of an Authentic Substrate


Bacteria in the genus Chlamydia are major human pathogens that cause an intracellular infection. A chlamydial protease, CPAF, has been proposed as an important virulence factor that cleaves or degrades at least 16 host proteins, thereby altering multiple cellular processes. We examined 11 published CPAF substrates and found that there was no detectable proteolysis when CPAF activity was inhibited during cell processing. We show that the reported proteolysis of these putative CPAF substrates was due to enzymatic activity in cell lysates rather than in intact cells. Nevertheless, Chlamydia-infected cells displayed Chlamydia-host interactions, such as Golgi reorganization, apoptosis resistance, and host cytoskeletal remodeling, that have been attributed to CPAF-dependent proteolysis of host proteins. Our findings suggest that other mechanisms may be responsible for these Chlamydia-host interactions, and raise concerns about all published CPAF substrates and the proposed roles of CPAF in chlamydial pathogenesis.


Vyšlo v časopise: CPAF: A Chlamydial Protease in Search of an Authentic Substrate. PLoS Pathog 8(8): e32767. doi:10.1371/journal.ppat.1002842
Kategorie: Research Article
prolekare.web.journal.doi_sk: https://doi.org/10.1371/journal.ppat.1002842

Souhrn

Bacteria in the genus Chlamydia are major human pathogens that cause an intracellular infection. A chlamydial protease, CPAF, has been proposed as an important virulence factor that cleaves or degrades at least 16 host proteins, thereby altering multiple cellular processes. We examined 11 published CPAF substrates and found that there was no detectable proteolysis when CPAF activity was inhibited during cell processing. We show that the reported proteolysis of these putative CPAF substrates was due to enzymatic activity in cell lysates rather than in intact cells. Nevertheless, Chlamydia-infected cells displayed Chlamydia-host interactions, such as Golgi reorganization, apoptosis resistance, and host cytoskeletal remodeling, that have been attributed to CPAF-dependent proteolysis of host proteins. Our findings suggest that other mechanisms may be responsible for these Chlamydia-host interactions, and raise concerns about all published CPAF substrates and the proposed roles of CPAF in chlamydial pathogenesis.


Zdroje

1. CDC 2011 Summary of notifiable diseases: United States, 2009. MMWR Morb Mortal Wkly Rep 58 1 100

2. SchachterJ 1999 Infection and disease epidemiology. StephensRS Chlamydia: Intracellular Biology, Pathogenesis, and Immunity Washington, D.C. American Society for Microbiology 139 169 editor pp

3. BurtonMJMabeyDC 2009 The global burden of trachoma: a review. PLoS Negl Trop Dis 3 e460

4. BlasiFTarsiaPAlibertiS 2009 Chlamydophila pneumoniae. Clin Microbiol Infect 15 29 35

5. CarabeoRAMeadDJHackstadtT 2003 Golgi-dependent transport of cholesterol to the Chlamydia trachomatis inclusion. Proc Natl Acad Sci U S A 100 6771 6776

6. HackstadtTRockeyDHeinzenRScidmoreM 1996 Chlamydia trachomatis interrupts an exocytic pathway to acquire endogenously synthesized sphingomyelin in transit from the Golgi apparatus to the plasma membrane. EMBO J 15 964 977

7. HackstadtTScidmoreMARockeyDD 1995 Lipid metabolism in Chlamydia trachomatis-infected cells: directed trafficking of Golgi-derived sphingolipids to the chlamydial inclusion. Proc Natl Acad Sci U S A 92 4877 4881

8. FanTLuHHuHShiLMcClartyGA 1998 Inhibition of apoptosis in Chlamydia-infected cells: blockade of mitochondrial cytochrome c release and caspase activation. J Exp Med 187 487 496

9. FischerSFSchwarzCVierJHackerG 2001 Characterization of antiapoptotic activities of Chlamydia pneumoniae in human cells. Infect Immun 69 7121 7129

10. RajalingamKAl-YounesHMullerAMeyerTFSzczepekAJ 2001 Epithelial cells infected with Chlamydophila pneumoniae (Chlamydia pneumoniae) are resistant to apoptosis. Infect Immun 69 7880 7888

11. ZhongG 2009 Killing me softly: chlamydial use of proteolysis for evading host defenses. Trends Microbiol 17 467 474

12. HuangZFengYChenDWuXHuangS 2008 Structural basis for activation and inhibition of the secreted Chlamydia protease CPAF. Cell Host Microbe 4 529 542

13. DongFZhongYArulanandamBZhongG 2005b Production of a proteolytically active protein, chlamydial protease/proteasome-like activity factor, by five different Chlamydia species. Infect Immun 73 1868 1872

14. HornMCollingroASchmitz-EsserSBeierCLPurkholdU 2004 Illuminating the evolutionary history of chlamydiae. Science 304 728 730

15. ZhongGFanPJiHDongFHuangY 2001 Identification of a chlamydial protease-like activity factor responsible for the degradation of host transcription factors. J Exp Med 193 935 942

16. ChristianJGHeymannJPaschenSAVierJSchauenburgL 2011 Targeting of a chlamydial protease impedes intracellular bacterial growth. PLoS Pathog 7 e1002283

17. HeuerDRejman LipinskiAMachuyNKarlasAWehrensA 2009 Chlamydia causes fragmentation of the Golgi compartment to ensure reproduction. Nature 457 731 735

18. PaschenSAChristianJGVierJSchmidtFWalchA 2008 Cytopathicity of Chlamydia is largely reproduced by expression of a single chlamydial protease. J Cell Biol 182 117 127

19. PirbhaiMDongFZhongYPanKZZhongG 2006 The secreted protease factor CPAF is responsible for degrading pro-apoptotic BH3-only proteins in Chlamydia trachomatis-infected cells. J Biol Chem 281 31495 31501

20. KumarYValdiviaRH 2008 Actin and intermediate filaments stabilize the Chlamydia trachomatis vacuole by forming dynamic structural scaffolds. Cell Host Microbe 4 159 169

21. ZhongGLiuLFanTFanPJiH 2000 Degradation of transcription factor RFX5 during the inhibition of both constitutive and interferon gamma-inducible major histocompatibility complex class I expression in Chlamydia-infected cells. J Exp Med 191 1525 1534

22. ChristianJVierJPaschenSAHackerG 2010 Cleavage of the NF-kappaB family protein p65/RelA by the chlamydial protease-like activity factor (CPAF) impairs proinflammatory signaling in cells infected with Chlamydiae. J Biol Chem 285 41320 41327

23. SunJSchoborgRV 2009 The host adherens junction molecule nectin-1 is degraded by chlamydial protease-like activity factor (CPAF) in Chlamydia trachomatis-infected genital epithelial cells. Microbes Infect 11 12 19

24. BellandRJZhongGCraneDDHoganDSturdevantD 2003 Genomic transcriptional profiling of the developmental cycle of Chlamydia trachomatis. Proc Natl Acad Sci U S A 100 8478 8483

25. HoldenPHortonWA 2009 Crude subcellular fractionation of cultured mammalian cell lines. BMC Res Notes 2 243

26. DickLRCruikshankAADestreeATGrenierLMcCormackTA 1997 Mechanistic studies on the inactivation of the proteasome by lactacystin in cultured cells. J Biol Chem 272 182 188

27. FenteanyGStandaertRFLaneWSChoiSCoreyEJ 1995 Inhibition of proteasome activities and subunit-specific amino-terminal threonine modification by lactacystin. Science 268 726 731

28. RajagopalanKVFridovichIHandlerP 1961 Competitive inhibition of enzyme activity by urea. J Biol Chem 236 1059 1065

29. TakizawaPAYucelJKVeitBFaulknerDJDeerinckT 1993 Complete vesiculation of Golgi membranes and inhibition of protein transport by a novel sea sponge metabolite, ilimaquinone. Cell 73 1079 1090

30. DongFSuHHuangYZhongYZhongG 2004c Cleavage of host keratin 8 by a Chlamydia-secreted protease. Infect Immun 72 3863 3868

31. BalsaraZRMisaghiSLafaveJNStarnbachMN 2006 Chlamydia trachomatis infection induces cleavage of the mitotic cyclin B1. Infect Immun 74 5602 5608

32. DongFPirbhaiMZhongYZhongG 2004a Cleavage-dependent activation of a Chlamydia-secreted protease. Mol Microbiol 52 1487 1494

33. ChenDChaiJHartPJZhongG 2009 Identifying catalytic residues in CPAF, a Chlamydia-secreted protease. Arch Biochem Biophys 485 16 23

34. ChenDLeiLFloresRHuangZWuZ 2010 Autoprocessing and self-activation of the secreted protease CPAF in Chlamydia-infected cells. Microb Pathog 49 164 173

35. DongFSharmaJXiaoYZhongYZhongG 2004b Intramolecular dimerization is required for the Chlamydia-secreted protease CPAF to degrade host transcriptional factors. Infect Immun 72 3869 3875

36. DongFPirbhaiMXiaoYZhongYWuY 2005a Degradation of the proapoptotic proteins Bik, Puma, and Bim with Bcl-2 domain 3 homology in Chlamydia trachomatis-infected cells. Infect Immun 73 1861 1864

37. JorgensenIBednarMMAminVDavisBKTingJP 2011 The Chlamydia protease CPAF regulates host and bacterial proteins to maintain pathogen vacuole integrity and promote virulence. Cell Host Microbe 10 21 32

38. LadSPLiJda Silva CorreiaJPanQGadwalS 2007 Cleavage of p65/RelA of the NF-kappaB pathway by Chlamydia. Proc Natl Acad Sci U S A 104 2933 2938

39. RajalingamKSharmaMLohmannCOswaldMThieckO 2008 Mcl-1 is a key regulator of apoptosis resistance in Chlamydia trachomatis-infected cells. PLoS One 3 e3102

40. RajalingamKSharmaMPalandNHurwitzRThieckO 2006 IAP-IAP complexes required for apoptosis resistance of C. trachomatis-infected cells. PLoS Pathog 2 e114

41. FischerUJanickeRUSchulze-OsthoffK 2003 Many cuts to ruin: a comprehensive update of caspase substrates. Cell Death Differ 10 76 100

42. CongYJupelliMGuentzelMNZhongGMurthyAK 2007 Intranasal immunization with chlamydial protease-like activity factor and CpG deoxynucleotides enhances protective immunity against genital Chlamydia muridarum infection. Vaccine 25 3773 3780

43. MurthyAKCongYMurpheyCGuentzelMNForsthuberTG 2006 Chlamydial protease-like activity factor induces protective immunity against genital chlamydial infection in transgenic mice that express the human HLA-DR4 allele. Infect Immun 74 6722 6729

44. SharmaJBosnicAMPiperJMZhongG 2004 Human antibody responses to a Chlamydia-secreted protease factor. Infect Immun 72 7164 7171

45. SkworTKandelRPBasraviSKhanASharmaB 2010 Characterization of humoral immune responses to chlamydial HSP60, CPAF, and CT795 in inflammatory and severe trachoma. Invest Ophthalmol Vis Sci 51 5128 5136

46. OssewaardeJde VriesABestebroerTAnguloA 1996 Application of a Mycoplasma group-specific PCR for monitoring decontamination of Mycoplasma-infected Chlamydia sp. strains. Appl Environ Microbiol 62 328 331

47. HarlowELaneD 1999 Using antibodies: a laboratory manual Cold Spring Harbor Laboratory Press pp 495

48. FischerSFVierJKirschnekSKlosAHessS 2004 Chlamydia inhibit host cell apoptosis by degradation of proapoptotic BH3-only proteins. J Exp Med 200 905 916

49. SunJKintnerJSchoborgRV 2008 The host adherens junction molecule nectin-1 is downregulated in Chlamydia trachomatis-infected genital epithelial cells. Microbiology 154 1290 1299

50. ZhongGFanTLiuL 1999 Chlamydia inhibits interferon gamma-inducible major histocompatibility complex class II expression by degradation of upstream stimulatory factor 1. J Exp Med 189 1931 1938

51. KawanaKQuayleAJFicarraMIbanaJAShenL 2007 CD1d degradation in Chlamydia trachomatis-infected epithelial cells is the result of both cellular and chlamydial proteasomal activity. J Biol Chem 282 7368 7375

52. YuHSchwarzerKForsterMKniemeyerOForsbach-BirkV 2010 Role of high-mobility group box 1 protein and poly(ADP-ribose) polymerase 1 degradation in Chlamydia trachomatis-induced cytopathicity. Infect Immun 78 3288 3297

53. RuppJGieffersJKlingerMvan ZandbergenGWraseR 2007 Chlamydia pneumoniae directly interferes with HIF-1alpha stabilization in human host cells. Cell Microbiol 9 2181 2191

Štítky
Hygiena a epidemiológia Infekčné lekárstvo Laboratórium

Článok vyšiel v časopise

PLOS Pathogens


2012 Číslo 8
Najčítanejšie tento týždeň
Najčítanejšie v tomto čísle
Kurzy

Zvýšte si kvalifikáciu online z pohodlia domova

Získaná hemofilie - Povědomí o nemoci a její diagnostika
nový kurz

Eozinofilní granulomatóza s polyangiitidou
Autori: doc. MUDr. Martina Doubková, Ph.D.

Všetky kurzy
Prihlásenie
Zabudnuté heslo

Zadajte e-mailovú adresu, s ktorou ste vytvárali účet. Budú Vám na ňu zasielané informácie k nastaveniu nového hesla.

Prihlásenie

Nemáte účet?  Registrujte sa

#ADS_BOTTOM_SCRIPTS#