Functional dissection of lipid and protein kinase signals of PIKfyve reveals the role of PtdIns 3,5-P2 production for endomembrane integrity - PubMed (original) (raw)
. 2002 Mar 15;277(11):9206-11.
doi: 10.1074/jbc.M108750200. Epub 2001 Nov 19.
Affiliations
- PMID: 11714711
- DOI: 10.1074/jbc.M108750200
Free article
Functional dissection of lipid and protein kinase signals of PIKfyve reveals the role of PtdIns 3,5-P2 production for endomembrane integrity
Ognian C Ikonomov et al. J Biol Chem. 2002.
Free article
Abstract
PIKfyve enzymatic activity is required in maintaining late endocytic membrane integrity. PIKfyve is a dual specificity enzyme that phosphorylates phosphatidylinositol (PtdIns) and PtdIns 3-P at the 5-hydroxyl and unidentified endogenous protein substrate(s). To determine which of these activities (lipid versus protein kinase activity) is responsible for endomembrane homeostasis we analyzed a double mutant PIKfyve(K1999E/K2000E). These substitutions in the putative lipid-substrate activation loop nearly completely abrogated the lipid kinase activity without any significant effect on the protein kinase activity of PIKfyve(K1999E/K2000E). Expression of PIKfyve(K1999E/K2000E) in COS cells induced a dramatic dominant-negative effect in the form of endomembrane swelling and vacuolation. In addition, the lipid-substrate specificity of PIKfyve was modified by introducing single mutations in Lys-1999 or Lys-2000. This yielded proteins with preferentially abrogated synthesis of PtdIns 5-P (PIKfyve(K2000E)) or PtdIns 3,5-P(2) (PIKfyve(K1999E)), of which only the PIKfyve(K1999E) mutant induced the characteristic endomembrane defects upon cell transfection. Furthermore, phosphoinositide microinjection into cells demonstrated a selective ability of PtdIns 3,5-P(2) to correct the endomembrane defects induced by the dominant-negative PIKfyve lipid kinase-deficient mutants. Thus, PtdIns 3,5-P(2) production by PIKfyve is crucial for endomembrane integrity, and Lys-1999 most likely directs the PIKfyve interactions with the 3-phosphate group in PtdIns 3-P.
Similar articles
- Mammalian cell morphology and endocytic membrane homeostasis require enzymatically active phosphoinositide 5-kinase PIKfyve.
Ikonomov OC, Sbrissa D, Shisheva A. Ikonomov OC, et al. J Biol Chem. 2001 Jul 13;276(28):26141-7. doi: 10.1074/jbc.M101722200. Epub 2001 Apr 2. J Biol Chem. 2001. PMID: 11285266 - Active vacuolar H+ ATPase and functional cycle of Rab5 are required for the vacuolation defect triggered by PtdIns(3,5)P2 loss under PIKfyve or Vps34 deficiency.
Compton LM, Ikonomov OC, Sbrissa D, Garg P, Shisheva A. Compton LM, et al. Am J Physiol Cell Physiol. 2016 Sep 1;311(3):C366-77. doi: 10.1152/ajpcell.00104.2016. Epub 2016 Jun 22. Am J Physiol Cell Physiol. 2016. PMID: 27335171 Free PMC article. - Functional dissociation between PIKfyve-synthesized PtdIns5P and PtdIns(3,5)P2 by means of the PIKfyve inhibitor YM201636.
Sbrissa D, Ikonomov OC, Filios C, Delvecchio K, Shisheva A. Sbrissa D, et al. Am J Physiol Cell Physiol. 2012 Aug 15;303(4):C436-46. doi: 10.1152/ajpcell.00105.2012. Epub 2012 May 23. Am J Physiol Cell Physiol. 2012. PMID: 22621786 Free PMC article. - PIKfyve: Partners, significance, debates and paradoxes.
Shisheva A. Shisheva A. Cell Biol Int. 2008 Jun;32(6):591-604. doi: 10.1016/j.cellbi.2008.01.006. Epub 2008 Jan 25. Cell Biol Int. 2008. PMID: 18304842 Free PMC article. Review. - PIKfyve and its Lipid products in health and in sickness.
Shisheva A. Shisheva A. Curr Top Microbiol Immunol. 2012;362:127-62. doi: 10.1007/978-94-007-5025-8_7. Curr Top Microbiol Immunol. 2012. PMID: 23086417 Review.
Cited by
- The synthetic TRPML1 agonist ML-SA1 rescues Alzheimer-related alterations of the endosomal-autophagic-lysosomal system.
Somogyi A, Kirkham ED, Lloyd-Evans E, Winston J, Allen ND, Mackrill JJ, Anderson KE, Hawkins PT, Gardiner SE, Waller-Evans H, Sims R, Boland B, O'Neill C. Somogyi A, et al. J Cell Sci. 2023 Mar 15;136(6):jcs259875. doi: 10.1242/jcs.259875. Epub 2023 Mar 21. J Cell Sci. 2023. PMID: 36825945 Free PMC article. - Identification and Utilization of a Chemical Probe to Interrogate the Roles of PIKfyve in the Lifecycle of β-Coronaviruses.
Drewry DH, Potjewyd FM, Bayati A, Smith JL, Dickmander RJ, Howell S, Taft-Benz S, Min SM, Hossain MA, Heise M, McPherson PS, Moorman NJ, Axtman AD. Drewry DH, et al. J Med Chem. 2022 Oct 13;65(19):12860-12882. doi: 10.1021/acs.jmedchem.2c00697. Epub 2022 Sep 16. J Med Chem. 2022. PMID: 36111834 Free PMC article. - NAADP-Dependent TPC Current.
Wang Q, Zhu MX. Wang Q, et al. Handb Exp Pharmacol. 2023;278:35-56. doi: 10.1007/164_2022_606. Handb Exp Pharmacol. 2023. PMID: 35902437 - PP2A-dependent TFEB activation is blocked by PIKfyve-induced mTORC1 activity.
Hasegawa J, Tokuda E, Yao Y, Sasaki T, Inoki K, Weisman LS. Hasegawa J, et al. Mol Biol Cell. 2022 Mar 1;33(3):ar26. doi: 10.1091/mbc.E21-06-0309. Epub 2022 Jan 12. Mol Biol Cell. 2022. PMID: 35020443 Free PMC article. - PIKfyve activity is required for lysosomal trafficking of tau aggregates and tau seeding.
Soares AC, Ferreira A, Mariën J, Delay C, Lee E, Trojanowski JQ, Moechars D, Annaert W, De Muynck L. Soares AC, et al. J Biol Chem. 2021 Jan-Jun;296:100636. doi: 10.1016/j.jbc.2021.100636. Epub 2021 Apr 6. J Biol Chem. 2021. PMID: 33831417 Free PMC article.
Publication types
MeSH terms
Substances
LinkOut - more resources
Full Text Sources
Other Literature Sources