Phosphorylation of a Myosin Motor by TgCDPK3 Facilitates Rapid Initiation of Motility during egress

English version České info

Toxoplasma gondii can cause severe disease and death in the immunocompromised and in those infected congenitally. Due to limitations of existing drugs there is a need for studying proteins that are unique and essential to the parasite. We recently established that TgCDPK3, a member of a family of calcium dependent protein kinase present in plants and some parasites but absent in human cells, regulates parasite egress from the host cell. While it has been hypothesized that TgCDPK3 controls rapid exit from the host by phosphorylating proteins needed for activating motility, the particular substrates of this kinase remained unknown. We have now applied an interaction trap system to identify the proteins that are modified by this kinase, which include a parasite motor protein Myosin A (TgMyoA). We show that TgCDPK3 specifically phosphorylates TgMyoA and this phosphorylation is important for parasite egress and motility.

Vyšlo v časopise: Phosphorylation of a Myosin Motor by TgCDPK3 Facilitates Rapid Initiation of Motility during egress. PLoS Pathog 11(11): e32767. doi:10.1371/journal.ppat.1005268
Kategorie: Research Article
prolekare.web.journal.doi_sk: https://doi.org/10.1371/journal.ppat.1005268

Souhrn

Zdroje

1. Luft BJ, Remington JS. Toxoplasmic encephalitis in AIDS. Clin Infect Dis. 1992;15(2):211–22. 1520757

2. Israelski DM, Remington JS. Toxoplasmosis in patients with cancer. Clin Infect Dis. 1993;17 Suppl 2:S423–35. 8274608

3. Slavin MA, Meyers JD, Remington JS, Hackman RC. Toxoplasma gondii infection in marrow transplant recipients: a 20 year experience. Bone Marrow Transplant. 1994;13(5):549–57. 8054907

4. Wong SY, Remington JS. Toxoplasmosis in pregnancy. Clin Infect Dis. 1994;18(6):853–61. 8086543

5. Black MW, Boothroyd JC. Lytic cycle of Toxoplasma gondii. Microbiol Mol Biol Rev. 2000;64(3):607–23. 10974128

6. Mehta S, Sibley LD. Actin depolymerizing factor controls actin turnover and gliding motility in Toxoplasma gondii. Molecular biology of the cell. 2011;22(8):1290–9. doi: 10.1091/mbc.E10-12-0939 21346192

7. Frenal K, Polonais V, Marq JB, Stratmann R, Limenitakis J, Soldati-Favre D. Functional dissection of the apicomplexan glideosome molecular architecture. Cell Host Microbe. 2010;8(4):343–57. Epub 2010/10/19. doi: 10.1016/j.chom.2010.09.002 20951968

8. Carruthers VB, Sibley LD. Mobilization of intracellular calcium stimulates microneme discharge in Toxoplasma gondii. Molecular microbiology. 1999;31(2):421–8. 10027960.

9. Andenmatten N, Egarter S, Jackson AJ, Jullien N, Herman JP, Meissner M. Conditional genome engineering in Toxoplasma gondii uncovers alternative invasion mechanisms. Nat Methods. 2013;10(2):125–7. Epub 012 Dec 23. doi: 10.1038/nmeth.2301 23263690

10. Kafsack BF, Pena JD, Coppens I, Ravindran S, Boothroyd JC, Carruthers VB. Rapid membrane disruption by a perforin-like protein facilitates parasite exit from host cells. Science (New York, NY. 2009;323(5913):530–3.

11. Nagamune K, Hicks LM, Fux B, Brossier F, Chini EN, Sibley LD. Abscisic acid controls calcium-dependent egress and development in Toxoplasma gondii. Nature. 2008;451(7175):207–10. doi: 10.1038/nature06478 18185591

12. Billker O, Lourido S, Sibley LD. Calcium-dependent signaling and kinases in apicomplexan parasites. Cell Host Microbe. 2009;5(6):612–22. Epub 2009/06/17. doi: 10.1016/j.chom.2009.05.017 19527888

13. Lourido S, Shuman J, Zhang C, Shokat KM, Hui R, Sibley LD. Calcium-dependent protein kinase 1 is an essential regulator of exocytosis in Toxoplasma. Nature. 2010;465(7296):359–62. Epub 2010/05/21. doi: 10.1038/nature09022 20485436

14. Garrison E, Treeck M, Ehret E, Butz H, Garbuz T, Oswald BP, et al. A Forward Genetic Screen Reveals that Calcium-dependent Protein Kinase 3 Regulates Egress in Toxoplasma. PLoS pathogens. 2012;8(11):e1003049. doi: 10.1371/journal.ppat.1003049 23209419

15. Lourido S, Tang K, Sibley LD. Distinct signalling pathways control Toxoplasma egress and host-cell invasion. The EMBO journal. 2012. Epub 2012/11/15.

16. McCoy JM, Whitehead L, van Dooren GG, Tonkin CJ. TgCDPK3 Regulates Calcium-Dependent Egress of Toxoplasma gondii from Host Cells. PLoS pathogens. 2012;8(12):e1003066. doi: 10.1371/journal.ppat.1003066 23226109

17. Black MW, Arrizabalaga G, Boothroyd JC. Ionophore-resistant mutants of Toxoplasma gondii reveal host cell permeabilization as an early event in egress. Molecular and cellular biology. 2000;20(24):9399–408. 11094090

18. Donald RG, Zhong T, Wiersma H, Nare B, Yao D, Lee A, et al. Anticoccidial kinase inhibitors: identification of protein kinase targets secondary to cGMP-dependent protein kinase. Molecular and biochemical parasitology. 2006;149(1):86–98. Epub 2006/06/13. 16765465

19. Sugi T, Kato K, Kobayashi K, Pandey K, Takemae H, Kurokawa H, et al. Molecular analyses of Toxoplasma gondii calmodulin-like domain protein kinase isoform 3. Parasitol Int. 2009;58(4):416–23. Epub 2009/08/25. doi: 10.1016/j.parint.2009.08.005 19699312

20. Treeck M, Sanders JL, Gaji RY, LaFavers KA, Child MA, Arrizabalaga G, et al. The Calcium-Dependent Protein Kinase 3 of Toxoplasma Influences Basal Calcium Levels and Functions beyond Egress as Revealed by Quantitative Phosphoproteome Analysis. PLoS pathogens. 2014;10(6):e1004197. Epub 2014/06/20. doi: 10.1371/journal.ppat.1004197 24945436

21. Anderson-White BR, Ivey FD, Cheng K, Szatanek T, Lorestani A, Beckers CJ, et al. A family of intermediate filament-like proteins is sequentially assembled into the cytoskeleton of Toxoplasma gondii. Cellular microbiology. 2011;13(1):18–31. Epub 2010/08/12. doi: 10.1111/j.1462-5822.2010.01514.x 20698859

22. Liu J, Wetzel L, Zhang Y, Nagayasu E, Ems-McClung S, Florens L, et al. Novel thioredoxin-like proteins are components of a protein complex coating the cortical microtubules of Toxoplasma gondii. Eukaryotic cell. 2013. Epub 2013/07/23.

23. Tang Q, Andenmatten N, Triana MA, Deng B, Meissner M, Moreno SN, et al. Calcium-dependent phosphorylation alters Class XIVa myosin function in the protozoan parasite Toxoplasma gondii. Molecular biology of the cell. 2014. Epub 2014/07/06.

24. Roux KJ, Kim DI, Raida M, Burke B. A promiscuous biotin ligase fusion protein identifies proximal and interacting proteins in mammalian cells. The Journal of cell biology. 2012;196(6):801–10. Epub 2012/03/14. doi: 10.1083/jcb.201112098 22412018

25. Kinoshita E, Kinoshita-Kikuta E, Koike T. Separation and detection of large phosphoproteins using Phos-tag SDS-PAGE. Nature protocols. 2009;4(10):1513–21. doi: 10.1038/nprot.2009.154 19798084

26. Tang Q, Andenmatten N, Hortua Triana MA, Deng B, Meissner M, Moreno SN, et al. Calcium-dependent phosphorylation alters class XIVa myosin function in the protozoan parasite Toxoplasma gondii. Molecular biology of the cell. 2014;25(17):2579–91. doi: 10.1091/mbc.E13-11-0648 24989796

27. Dephoure N, Gould KL, Gygi SP, Kellogg DR. Mapping and analysis of phosphorylation sites: a quick guide for cell biologists. Molecular biology of the cell. 2013;24(5):535–42. doi: 10.1091/mbc.E12-09-0677 23447708

28. Hunter T. Why nature chose phosphate to modify proteins. Philos Trans R Soc Lond B Biol Sci. 2012;367(1602):2513–6. doi: 10.1098/rstb.2012.0013 22889903

29. Egarter S, Andenmatten N, Jackson AJ, Whitelaw JA, Pall G, Black JA, et al. The Toxoplasma Acto-MyoA motor complex is important but not essential for gliding motility and host cell invasion. PloS one. 2014;9(3):e91819. doi: 10.1371/journal.pone.0091819 24632839

30. Meissner M, Schluter D, Soldati D. Role of Toxoplasma gondii myosin A in powering parasite gliding and host cell invasion. Science (New York, NY. 2002;298(5594):837–40.

31. Pomel S, Luk FC, Beckers CJ. Host cell egress and invasion induce marked relocations of glycolytic enzymes in Toxoplasma gondii tachyzoites. PLoS pathogens. 2008;4(10):e1000188. Epub 2008/10/25. doi: 10.1371/journal.ppat.1000188 18949028

32. Hakansson S, Morisaki H, Heuser J, Sibley LD. Time-lapse video microscopy of gliding motility in Toxoplasma gondii reveals a novel, biphasic mechanism of cell locomotion. Molecular biology of the cell. 1999;10(11):3539–47. 10564254

33. Hu K, Johnson J, Florens L, Fraunholz M, Suravajjala S, DiLullo C, et al. Cytoskeletal components of an invasion machine—the apical complex of Toxoplasma gondii. PLoS pathogens. 2006;2(2):e13. 16518471

34. Gubbels MJ, Vaishnava S, Boot N, Dubremetz JF, Striepen B. A MORN-repeat protein is a dynamic component of the Toxoplasma gondii cell division apparatus. Journal of cell science. 2006;119(Pt 11):2236–45. Epub 2006/05/11. 16684814

35. Heaslip AT, Dzierszinski F, Stein B, Hu K. TgMORN1 is a key organizer for the basal complex of Toxoplasma gondii. PLoS pathogens. 2010;6(2):e1000754. doi: 10.1371/journal.ppat.1000754 20140195

36. Keeley A, and Soldati D. The glideosome: a molecular machine powering motility and host-cell invasion by Apicomplexa. Trends in Cell Bio. 2004;14(10):528–32.

37. Nebl T, Prieto JH, Kapp E, Smith BJ, Williams MJ, Yates JR3rd, et al. Quantitative in vivo analyses reveal calcium-dependent phosphorylation sites and identifies a novel component of the Toxoplasma invasion motor complex. PLoS pathogens. 2011;7(9):e1002222. Epub 2011/10/08. doi: 10.1371/journal.ppat.1002222 21980283

38. Gilk SD, Gaskins E, Ward GE, Beckers CJ. GAP45 phosphorylation controls assembly of the Toxoplasma myosin XIV complex. Eukaryotic cell. 2009;8(2):190–6. Epub 2008/12/03. doi: 10.1128/EC.00201-08 19047362

39. Foth BJ, Goedecke MC, Soldati D. New insights into myosin evolution and classification. Proc Natl Acad Sci U S A. 2006;103(10):3681–6. 16505385

40. Heintzelman MB, Schwartzman JD. Myosin diversity in Apicomplexa. The Journal of parasitology. 2001;87(2):429–32. 11318578

41. Herm-Gotz A, Weiss S, Stratmann R, Fujita-Becker S, Ruff C, Meyhofer E, et al. Toxoplasma gondii myosin A and its light chain: a fast, single-headed, plus-end-directed motor. The EMBO journal. 2002;21(9):2149–58. 11980712

42. Bookwalter CS, Kelsen A, Leung JM, Ward GE, Trybus KM. A Toxoplasma gondii class XIV myosin, expressed in Sf9 cells with a parasite co-chaperone, requires two light chains for fast motility. The Journal of biological chemistry. 2014;289(44):30832–41. Epub 2014/09/19. doi: 10.1074/jbc.M114.572453 25231988

43. Gaji RY, Checkley L, Reese M, Ferdig MT, Arrizabalaga G. Expression of the essential kinase PfCDPK1 from Plasmodium falciparum in Toxoplasma gondii facilitates the discovery of novel antimalarial drugs. Antimicrobial agents and chemotherapy. 2014. Epub 2014/02/20.

44. Soldati D, Boothroyd JC. Transient transfection and expression in the obligate intracellular parasite Toxoplasma gondii. Science (New York, NY. 1993;260(5106):349–52.

45. Kim K, Soldati D, Boothroyd JC. Gene replacement in Toxoplasma gondii with chloramphenicol acetyltransferase as selectable marker. Science (New York, NY. 1993;262(5135):911–4.

46. Chen AL, Kim EW, Toh JY, Vashisht AA, Rashoff AQ, Van C, et al. Novel components of the Toxoplasma inner membrane complex revealed by BioID. mBio. 2015;6(1):e02357–14. doi: 10.1128/mBio.02357-14 25691595

47. Ashpole NM, Song W, Brustovetsky T, Engleman EA, Brustovetsky N, Cummins TR, et al. Calcium/calmodulin-dependent protein kinase II (CaMKII) inhibition induces neurotoxicity via dysregulation of glutamate/calcium signaling and hyperexcitability. The Journal of biological chemistry. 2012;287(11):8495–506. doi: 10.1074/jbc.M111.323915 22253441

48. Gaji RY, Flammer HP, Carruthers VB. Forward targeting of Toxoplasma gondii proproteins to the micronemes involves conserved aliphatic amino acids. Traffic. 2011;12(7):840–53. doi: 10.1111/j.1600-0854.2011.01192.x 21438967