Leishmania donovani 90 kD Heat Shock Protein - Impact of Phosphosites on Parasite Fitness, Infectivity and Casein Kinase Affinity.

Publisher: Nature Publishing Group Country of Publication: England NLM ID: 101563288 Publication Model: Electronic Cited Medium: Internet ISSN: 2045-2322 (Electronic) Linking ISSN: 20452322 NLM ISO Abbreviation: Sci Rep Subsets: MEDLINE

Original Publication: London : Nature Publishing Group, copyright 2011-

Casein Kinases/*metabolism ; HSP90 Heat-Shock Proteins/*metabolism ; Leishmania donovani/*metabolism ; Leishmania donovani/*pathogenicity; Amino Acid Sequence ; Casein Kinases/genetics ; HSP90 Heat-Shock Proteins/genetics ; Leishmania donovani/genetics ; Microscopy, Fluorescence ; Molecular Sequence Data ; Mutagenesis ; Mutation ; Phosphorylation ; Signal Transduction/genetics

Leishmania parasites are thought to control protein activity at the post-translational level, e.g. by protein phosphorylation. In the pathogenic amastigote, the mammalian stage of Leishmania parasites, heat shock proteins show increased phosphorylation, indicating a role in stage-specific signal transduction. Here we investigate the impact of phosphosites in the L. donovani heat shock protein 90. Using a chemical knock-down/genetic complementation approach, we mutated 11 confirmed or presumed phosphorylation sites and assessed the impact on overall fitness, morphology and in vitro infectivity. Most phosphosite mutations affected the growth and morphology of promastigotes in vitro, but with one exception, none of the phosphorylation site mutants had a selective impact on the in vitro infection of macrophages. Surprisingly, aspartate replacements mimicking the negative charge of phosphorylated serines or threonines had mostly negative impacts on viability and infectivity. HSP90 is a substrate for casein kinase 1.2-catalysed phosphorylation in vitro. While several putative phosphosite mutations abrogated casein kinase 1.2 activity on HSP90, only Ser 289 could be identified as casein kinase target by mass spectrometry. In summary, our data show HSP90 as a downstream client of phosphorylation-mediated signalling in an organism that depends on post-transcriptional gene regulation.

Proteomics. 2011 Sep;11(17):3572-7. (PMID: 21751374)
Proteomics. 2008 Jan;8(2):350-63. (PMID: 18203260)
Cell Stress Chaperones. 2015 Jul;20(4):673-85. (PMID: 25948161)
Cell. 1994 Dec 30;79(7):1129-32. (PMID: 8001149)
Acta Trop. 1979 Sep;36(3):289-92. (PMID: 43092)
Protein Expr Purif. 1994 Apr;5(2):133-7. (PMID: 8054844)
FEBS Lett. 1991 Oct 21;291(2):359-62. (PMID: 1936287)
Cell Stress Chaperones. 2009 Jan;14(1):83-94. (PMID: 18636345)
Mol Cell Biol. 1995 Jul;15(7):3917-25. (PMID: 7791797)
J Proteomics Bioinform. 2011;4(2):22-35. (PMID: 21720494)
Nat Rev Cancer. 2005 Oct;5(10):761-72. (PMID: 16175177)
Cell Stress Chaperones. 1998 Jun;3(2):100-8. (PMID: 9672245)
Annu Rev Biochem. 2006;75:271-94. (PMID: 16756493)
Eukaryot Cell. 2002 Dec;1(6):936-43. (PMID: 12477794)
Nat Commun. 2017 May 24;8:15328. (PMID: 28537252)
J Eukaryot Microbiol. 2016 Nov;63(6):823-833. (PMID: 27216143)
Med Microbiol Immunol. 2016 Oct;205(5):449-58. (PMID: 27283847)
Nature. 1970 Aug 15;227(5259):680-5. (PMID: 5432063)
J Cell Biochem. 2009 Feb 15;106(3):399-408. (PMID: 19115251)
Mol Cell Biol. 1998 Sep;18(9):4949-60. (PMID: 9710578)
Antimicrob Agents Chemother. 2014;58(3):1501-15. (PMID: 24366737)
Int J Parasitol. 2009 Apr;39(5):541-6. (PMID: 19028498)
Exp Parasitol. 1987 Oct;64(2):147-56. (PMID: 3653336)
J Biol Chem. 2001 Dec 21;276(51):47922-9. (PMID: 11598129)
J Struct Biol. 2011 Aug;175(2):224-9. (PMID: 21605682)
J Cell Sci. 2014 Nov 1;127(Pt 21):4762-73. (PMID: 25179594)
Mol Microbiol. 2016 Jun;100(6):931-44. (PMID: 26991545)
Antimicrob Agents Chemother. 2004 Aug;48(8):3033-42. (PMID: 15273118)
Mol Cell. 2012 Aug 10;47(3):434-43. (PMID: 22727666)
Proc Natl Acad Sci U S A. 1994 Aug 30;91(18):8324-8. (PMID: 8078881)
Rapid Commun Mass Spectrom. 2007;21(22):3635-45. (PMID: 17939157)
J Biol Chem. 2001 Aug 31;276(35):32822-7. (PMID: 11438552)
Biochim Biophys Acta. 2012 Mar;1823(3):648-55. (PMID: 21856339)
Annu Rev Genet. 1988;22:631-77. (PMID: 2853609)
Int J Parasitol. 2009 Oct;39(12):1289-303. (PMID: 19445946)
Proc Natl Acad Sci U S A. 2003 Sep 2;100(18):10193-200. (PMID: 12925738)
Biochim Biophys Acta. 2000 Apr 25;1491(1-3):65-74. (PMID: 10760571)
Nature. 1998 Nov 26;396(6709):336-42. (PMID: 9845070)
Biochem J. 1995 Aug 15;310 ( Pt 1):225-32. (PMID: 7646449)
Proc Natl Acad Sci U S A. 2010 May 4;107(18):8381-6. (PMID: 20404152)
EMBO J. 1999 Feb 1;18(3):754-62. (PMID: 9927435)
Cell Stress Chaperones. 2010 Jul;15(4):443-55. (PMID: 19953351)
Mol Cell Biol. 2006 Nov;26(22):8385-95. (PMID: 16982694)
Gene. 2015 Oct 1;570(1):8-16. (PMID: 26071189)
Proteomics. 2003 Sep;3(9):1811-29. (PMID: 12973740)
Science. 1984 Mar 30;223(4643):1417-9. (PMID: 6701528)
Oncotarget. 2011 May;2(5):407-17. (PMID: 21576760)
FASEB J. 2008 Feb;22(2):590-602. (PMID: 17884972)
Front Oncol. 2014 May 19;4:96. (PMID: 24904820)
Plant Mol Biol. 1991 Jul;17(1):73-82. (PMID: 1907872)
Mol Cell. 2011 Mar 18;41(6):672-81. (PMID: 21419342)
J Biomed Biotechnol. 2010;2010:525241. (PMID: 20169133)
J Proteome Res. 2010 Jan;9(1):393-403. (PMID: 19888749)
IUBMB Life. 1999 Oct;48(4):425-8. (PMID: 10632573)
Cell Stress Chaperones. 2017 Sep;22(5):729-742. (PMID: 28455612)
FASEB J. 2011 Feb;25(2):515-25. (PMID: 20952481)
mSystems. 2018 Nov 20;3(6):. (PMID: 30505948)
Biochem J. 2001 May 1;355(Pt 3):609-15. (PMID: 11311121)
Mol Cell. 2014 Jan 23;53(2):317-29. (PMID: 24462205)
Eukaryot Cell. 2003 Oct;2(5):830-40. (PMID: 14555465)
Biol Pharm Bull. 2008 Feb;31(2):193-200. (PMID: 18239272)
Proc Natl Acad Sci U S A. 1999 Jan 5;96(1):109-14. (PMID: 9874780)
Mol Endocrinol. 1999 Sep;13(9):1435-48. (PMID: 10478836)
Int J Parasitol. 2008 Oct;38(12):1411-23. (PMID: 18457839)
Mol Biol Cell. 2001 Nov;12(11):3307-16. (PMID: 11694568)
Mol Biochem Parasitol. 1995 Mar;70(1-2):107-18. (PMID: 7637691)
J Cell Sci. 2010 Mar 15;123(Pt 6):842-52. (PMID: 20159964)
Nat Struct Mol Biol. 2011 Jan;18(1):61-6. (PMID: 21170051)
Parasit Vectors. 2014 May 20;7:234. (PMID: 24886176)
PLoS One. 2012;7(5):e35671. (PMID: 22693548)
Proteomics. 2007 Aug;7(15):2553-6. (PMID: 17610305)
J Biol Chem. 2003 May 16;278(20):18336-45. (PMID: 12584193)
Mol Endocrinol. 1996 Jun;10(6):705-12. (PMID: 8776730)
J Biol Chem. 1998 Mar 13;273(11):6488-94. (PMID: 9497383)
Mol Biochem Parasitol. 1997 Nov;89(2):179-93. (PMID: 9364964)
Trends Biochem Sci. 2000 Dec;25(12):596-601. (PMID: 11116185)
Int J Parasitol. 2002 Aug;32(9):1085-93. (PMID: 12117491)
Nat Biotechnol. 2006 Oct;24(10):1285-92. (PMID: 16964243)
Mol Cell Biol. 2009 Dec;29(24):6401-12. (PMID: 19805514)
Mol Biochem Parasitol. 2011 Apr;176(2):98-108. (PMID: 21195115)
Cell Microbiol. 2015 Oct;17(10):1405-12. (PMID: 25939677)
Biochem Pharmacol. 1998 Sep 15;56(6):675-82. (PMID: 9751071)
Cell Cycle. 2010 Jun 15;9(12):2310-6. (PMID: 20519952)
EMBO J. 1998 May 1;17(9):2619-28. (PMID: 9564044)
Sci Rep. 2017 Aug 31;7(1):10202. (PMID: 28860596)
J Cell Sci. 2004 Feb 15;117(Pt 5):701-10. (PMID: 14754904)
Genome Biol. 2008;9(2):R35. (PMID: 18282296)
Cell Microbiol. 2015 May;17(5):632-8. (PMID: 25801803)
Cell Microbiol. 2013 Apr;15(4):585-600. (PMID: 23107115)
Mol Cell Proteomics. 2014 Jul;13(7):1787-99. (PMID: 24741111)
J Immunol. 2010 Nov 1;185(9):5011-22. (PMID: 20881185)
Kinetoplastid Biol Dis. 2005 Apr 29;4(1):3. (PMID: 15862128)
Oncogene. 2013 Jun 20;32(25):3101-10. (PMID: 22824801)
Int J Parasitol. 2007 Aug;37(10):1053-62. (PMID: 17548090)
Mol Biochem Parasitol. 2005 May;141(1):99-108. (PMID: 15811531)
Genome Res. 2002 Aug;12(8):1231-45. (PMID: 12176931)
EMBO J. 2002 Apr 15;21(8):1881-8. (PMID: 11953307)
mBio. 2015 Jul 21;6(4):e00861. (PMID: 26199327)
Cancer Res. 2000 Jul 15;60(14):3940-6. (PMID: 10919672)

0 (HSP90 Heat-Shock Proteins)
EC 2.7.11.1 (Casein Kinases)

Date Created: 20190327 Date Completed: 20201019 Latest Revision: 20240715

20240715

PMC6434042

10.1038/s41598-019-41640-0

30911045