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Zhizhuang Joe Zhao - One of the best experts on this subject based on the ideXlab platform.
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Characterization and functional studies of a FYVE Domain-containing phosphatase in C. elegans.
Journal of cellular biochemistry, 2008Co-Authors: Fenghua Zeng, Lirong Teng, Zhizhuang Joe ZhaoAbstract:The myotubularin (MTM) enzymes are phosphatidylinositol 3-phosphate (PI3P) and phosphatidylinositol 3,5-bisphosphate phosphatases. Mutation of MTM1, the founder member of this family, is responsible for X-linked myotubular myopathy in humans. Here, we have isolated and characterized a Caenorhabditis elegans homology of the enzymes designated ceMTM3. ceMTM3 preferably dephosphorylates PI3P and contains a FYVE lipid-binding Domain at its C-terminus which binds PI3P. Immunoblotting analyses revealed that the enzyme is expressed during the early development and adulthood of the animal. Immunofluorescent staining revealed predominant expression of the enzyme in eggs and muscles. Knockdown of the enzyme by using feeding-based RNA interference resulted in an increased level of PI3P and caused severe impairment of body movement of the worms at their post-reproductive ages and significantly shortened their lifespan. This study thus reveals an important role of the MTM phosphatases in maintaining muscle function, which may have clinical implications in prevention and treatment of sarcopenia.
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FYVE-DSP2, a FYVE Domain-containing dual specificity protein phosphatase that dephosphorylates phosphotidylinositol 3-phosphate
Experimental Cell Research, 2001Co-Authors: Runxiang Zhao, Ying Qi, Jie Chen, Zhizhuang Joe ZhaoAbstract:Abstract We have recently isolated FYVE-DSP1, a FYVE Domain-containing dual specificity protein phosphatase (R. Zhao, Y. Qi, and Z. J. Zhao, Biochem. Biophys. Res. Commun. 270, 222–229 (2000)). Here, we report a novel isozyme that we designated FYVE-DSP2. FYVE-2 contains a single FYVE Domain at the C-terminus, and it shares ∼47% overall sequence identity with FYBE-DSP1. Genomic sequence analyses revealed that the FYVE-DSP1 and FYVE-DSP2 genes share similar intron/exon organization. They are localizedon human chromosome 22q12 and chromosome 17, respectively. Like FYVE-DSP1, recombinant FYVE-DSP2 dephosphorylated low-molecular-weight phosphatase substrate para-nitrophenylphosphate, and its activity was inhibited by sodium vanadate. More importantly, our study also revealed that both FYVE-DSP1 and FYVE-DSP2 efficiently and specifically dephosphorylated phosphotidylinositol 3-phosphate. Subcellular fractionation demonstrated partition of FYVE-DSP1 and FYVE-DSP2 in membrane fractions, and immunofluorescent cell staining showed perinuclear localization of the enzymes. FYVE-DSP2 is expressed in many human tissues with an alternatively spliced isoform expressed in the kidney. Together with two homologous hypothetical proteins found in Caenorhabditis elegans and Drosophila, FYVE-DSP1 and FYVE-DSP2 form a subfamilyof phosphatases that may have an importantrole in cellular processes.
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FYVE-DSP2, a FYVE Domain-containing dual specificity protein phosphatase that dephosphorylates phosphotidylinositol 3-phosphate
Experimental cell research, 2001Co-Authors: Runxiang Zhao, Jie Chen, Zhizhuang Joe ZhaoAbstract:We have recently isolated FYVE-DSP1, a FYVE Domain-containing dual specificity protein phosphatase (R. Zhao, Y. Qi, and Z. J. Zhao, Biochem. Biophys. Res. Commun. 270, 222--229 (2000)). Here, we report a novel isozyme that we designated FYVE-DSP2. FYVE-2 contains a single FYVE Domain at the C-terminus, and it shares approximately 47% overall sequence identity with FYBE-DSP1. Genomic sequence analyses revealed that the FYVE-DSP1 and FYVE-DSP2 genes share similar intron/exon organization. They are localizedon human chromosome 22q12 and chromosome 17, respectively. Like FYVE-DSP1, recombinant FYVE-DSP2 dephosphorylated low-molecular-weight phosphatase substrate para-nitrophenylphosphate, and its activity was inhibited by sodium vanadate. More importantly, our study also revealed that both FYVE-DSP1 and FYVE-DSP2 efficiently and specifically dephosphorylated phosphotidylinositol 3-phosphate. Subcellular fractionation demonstrated partition of FYVE-DSP1 and FYVE-DSP2 in membrane fractions, and immunofluorescent cell staining showed perinuclear localization of the enzymes. FYVE-DSP2 is expressed in many human tissues with an alternatively spliced isoform expressed in the kidney. Together with two homologous hypothetical proteins found in Caenorhabditis elegans and Drosophila, FYVE-DSP1 and FYVE-DSP2 form a subfamilyof phosphatases that may have an importantrole in cellular processes.
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FYVE-DSP1, a dual-specificity protein phosphatase containing an FYVE Domain.
Biochemical and biophysical research communications, 2000Co-Authors: Runxiang Zhao, Zhizhuang Joe ZhaoAbstract:Dual-specificity protein phosphatases (DSPs) dephosphorylate proteins at Ser/Thr and Tyr. FYVE Domain is a double zinc finger motif which specifically binds phosphatidylinositol(3)-phosphate. Here, we report a novel dual specificity phosphatase that contains a FYVE Domain at the C-terminus. We designate the protein FYVE-DSP1. Molecular cloning yielded three isoforms of the enzyme presumably derived from alternate RNA splicing. Sequence alignment revealed that the catalytic phosphatase Domain of FYVE-DSP1 closely resembled that of myotubularin, while its FYVE Domain has all the conserved amino acid residues found in other proteins of the same family. Recombinant FYVE-DSP1 is partitioned in both cytosolic and membrane fractions. It dephosphorylates proteins phosphorylated on Ser, Thr, and Tyr residues and low molecular weight phosphatase substrate para-nitrophenylphosphate. It shows typical characteristics of other DSPs and protein tyrosine phosphatases (PTPs). These include inhibition by sodium vanadate and pervanadate, pH dependency, and inactivation by mutation of the key cysteinyl residue at the phosphatase signature motif. Finally, PCR analyses demonstrated that FYVE-DSP1 is widely distributed in human tissues but different spliced forms expressed differently.
Margaret S Robinson - One of the best experts on this subject based on the ideXlab platform.
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rag gtpases and phosphatidylinositol 3 phosphate mediate recruitment of the ap 5 spg11 spg15 complex
Journal of Cell Biology, 2021Co-Authors: Jennifer Hirst, G Hesketh, Anneclaude Gingras, Margaret S RobinsonAbstract:Adaptor protein complex 5 (AP-5) and its partners, SPG11 and SPG15, are recruited onto late endosomes and lysosomes. Here we show that recruitment of AP-5/SPG11/SPG15 is enhanced in starved cells and occurs by coincidence detection, requiring both phosphatidylinositol 3-phosphate (PI3P) and Rag GTPases. PI3P binding is via the SPG15 FYVE Domain, which, on its own, localizes to early endosomes. GDP-locked RagC promotes recruitment of AP-5/SPG11/SPG15, while GTP-locked RagA prevents its recruitment. Our results uncover an interplay between AP-5/SPG11/SPG15 and the mTORC1 pathway and help to explain the phenotype of AP-5/SPG11/SPG15 deficiency in patients, including the defect in autophagic lysosome reformation.
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Rag GTPases and phosphatidylinositol 3-phosphate mediate recruitment of the AP-5/SPG11/SPG15 complex.
The Journal of cell biology, 2021Co-Authors: Jennifer Hirst, Anneclaude Gingras, Geoffrey G Hesketh, Margaret S RobinsonAbstract:Adaptor protein complex 5 (AP-5) and its partners, SPG11 and SPG15, are recruited onto late endosomes and lysosomes. Here we show that recruitment of AP-5/SPG11/SPG15 is enhanced in starved cells and occurs by coincidence detection, requiring both phosphatidylinositol 3-phosphate (PI3P) and Rag GTPases. PI3P binding is via the SPG15 FYVE Domain, which, on its own, localizes to early endosomes. GDP-locked RagC promotes recruitment of AP-5/SPG11/SPG15, while GTP-locked RagA prevents its recruitment. Our results uncover an interplay between AP-5/SPG11/SPG15 and the mTORC1 pathway and help to explain the phenotype of AP-5/SPG11/SPG15 deficiency in patients, including the defect in autophagic lysosome reformation.
Tatiana G. Kutateladze - One of the best experts on this subject based on the ideXlab platform.
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Membrane insertion of the FYVE Domain is modulated by pH
Proteins, 2009Co-Authors: Mohsin Vora, Rachel M. Haney, Grigory S. Filonov, Catherine A. Musselman, Christopher G. Burd, Andrei G. Kutateladze, Vladislav V. Verkhusha, Robert V. Stahelin, Tatiana G. KutateladzeAbstract:The FYVE Domain associates with phosphatidylinositol 3-phosphate [PtdIns(3)P] in membranes of early endosomes and penetrates bilayers. Here, we detail principles of membrane anchoring and show that the FYVE Domain insertion into PtdIns(3)P-enriched membranes and membrane-mimetics is substantially increased in acidic conditions. The EEA1 FYVE Domain binds to POPC/POPE/PtdIns(3)P vesicles with a Kd of 49 nM at pH 6.0, however associates ~24 fold weaker at pH 8.0. The decrease in the affinity is primarily due to much faster dissociation of the protein from the bilayers in basic media. Lowering the pH enhances the interaction of the Hrs, RUFY1, Vps27p and WDFY1 FYVE Domains with PtdIns(3)P-containing membranes in vitro and in vivo, indicating that pH-dependency is a general function of the FYVE finger family. The PtdIns(3)P binding and membrane insertion of the FYVE Domain is modulated by the two adjacent His residues of the R(R/K)HHCRXCG signature motif. Mutation of either His residue abolishes the pH-sensitivity. Both protonation of the His residues and nonspecific electrostatic contacts stabilize the FYVE Domain in the lipid-bound form, promoting its penetration and increasing the membrane residence time.
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Synthesis and Molecular Recognition of Phosphatidylinositol-3-methylenephosphate
Organic Letters, 2006Co-Authors: Joanna Gajewiak, Tatiana G. Kutateladze, Y. Xu, Glenn D PrestwichAbstract:Phosphatidylinositol-3-phosphate (PtdIns(3)P) is a spatial regulator of vesicular trafficking and other vital cellular processes. We describe the asymmetric total synthesis of a metabolically stabilized analogue, phosphatidylinositol-3-methylenephosphate (PtdIns(3)MP) from a differentially protected myo-inositol. NMR studies of PtdIns(3)MP bound to the 15N-labeled FYVE Domain showed significant 1H and 15N chemical shift changes relative to the unliganded protein.
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Synthesis and molecular recognition of phosphatidylinositol-3-methylenephosphate.
Organic letters, 2006Co-Authors: Joanna Gajewiak, Tatiana G. Kutateladze, Stephanie A. Lee, Glenn D PrestwichAbstract:[reaction: see text] Phosphatidylinositol-3-phosphate (PtdIns(3)P) is a spatial regulator of vesicular trafficking and other vital cellular processes. We describe the asymmetric total synthesis of a metabolically stabilized analogue, phosphatidylinositol-3-methylenephosphate (PtdIns(3)MP) from a differentially protected myo-inositol. NMR studies of PtdIns(3)MP bound to the (15)N-labeled FYVE Domain showed significant (1)H and (15)N chemical shift changes relative to the unliganded protein.
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Phosphatidylinositol 3-phosphate recognition and membrane docking by the FYVE Domain.
Biochimica et biophysica acta, 2006Co-Authors: Tatiana G. KutateladzeAbstract:The FYVE Domain is a small zinc binding module that recognizes phosphatidylinositol 3-phosphate [PtdIns(3)P], a phospholipid enriched in membranes of early endosomes and other endocytic vesicles. It is usually present as a single module or rarely as a tandem repeat in eukaryotic proteins involved in a variety of biological processes including endo- and exocytosis, membrane trafficking and phosphoinositide metabolism. A number of FYVE Domain-containing proteins are recruited to endocytic membranes through the specific interaction of their FYVE Domains with PtdIns(3)P. Structures and PtdIns(3)P binding modes of several FYVE Domains have recently been characterized, shedding light on the molecular basis underlying multiple cellular functions of these proteins. Here, structural and functional aspects and the current mechanism of the multivalent membrane anchoring by monomeric or dimeric FYVE Domain are reviewed. This mechanism involves stereospecific recognition of PtdIns(3)P that is facilitated by non-specific electrostatic contacts and modulated by the histidine switch, and is accompanied by hydrophobic insertion. Contributions of each component to the FYVE Domain specificity and affinity for PtdIns(3)P-containing membranes are discussed.
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Targeting of the FYVE Domain to endosomal membranes is regulated by a histidine switch.
Proceedings of the National Academy of Sciences of the United States of America, 2005Co-Authors: Stephanie A. Lee, Christopher G. Burd, Vladislav V. Verkhusha, Michael Overduin, Matthew L. Cheever, Rosemary Eyeson, Jinming Geng, Tatiana G. KutateladzeAbstract:Specific recognition of phosphatidylinositol 3-phosphate [PtdIns(3)P] by the FYVE Domain targets cytosolic proteins to endosomal membranes during key signaling and trafficking events within eukaryotic cells. Here, we show that this membrane targeting is regulated by the acidic cellular environment. Lowering the cytosolic pH enhances PtdIns(3)P affinity of the FYVE Domain, reinforcing the anchoring of early endosome antigen 1 (EEA1) to endosomal membranes. Reversibly, increasing the pH disrupts phosphoinositide binding and leads to cytoplasmic redistribution of EEA1. pH dependency is due to a pair of conserved His residues, the successive protonation of which is required for PtdIns(3)P head group recognition as revealed by NMR. Substitution of the His residues abolishes PtdIns(3)P binding by the FYVE Domain in vitro and in vivo. Another PtdIns(3)P-binding module, the PX Domain of Vam7 and p40phox is shown to be pH-independent. This provides the fundamental functional distinction between the two phosphoinositide-recognizing Domains. The presented mode of FYVE regulation establishes the unique function of FYVE proteins as low pH sensors of PtdIns(3)P and reveals the critical role of the histidine switch in targeting of these proteins to endosomal membranes.
James H Hurley - One of the best experts on this subject based on the ideXlab platform.
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crystal structure of a phosphatidylinositol 3 phosphate specific membrane targeting motif the FYVE Domain of vps27p
Cell, 1999Co-Authors: Saurav Misra, James H HurleyAbstract:Abstract Phosphatidylinositol 3-phosphate regulates membrane trafficking and signaling pathways by interacting with the FYVE Domains of target proteins. The 1.15 A structure of the Vps27p FYVE Domain reveals two antiparallel β sheets and an α helix stabilized by two Zn 2+ -binding clusters. The core secondary structures are similar to a rabphilin-3A Zn 2+ -binding Domain and to the C1 and LIM Domains. Phosphatidylinositol 3-phosphate binds to a pocket formed by the (R/K)(R/K)HHCR motif. A lattice contact shows how anionic ligands can interact with the phosphatidylinositol 3-phosphate-binding site. The tip of the FYVE Domain has basic and hydrophobic surfaces positioned so that nonspecific interactions with the phospholipid bilayer can abet specific binding to phosphatidylinositol 3-phosphate.
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Crystal structure of a phosphatidylinositol 3-phosphate-specific membrane-targeting motif, the FYVE Domain of Vps27p.
Cell, 1999Co-Authors: Saurav Misra, James H HurleyAbstract:Phosphatidylinositol 3-phosphate regulates membrane trafficking and signaling pathways by interacting with the FYVE Domains of target proteins. The 1.15 A structure of the Vps27p FYVE Domain reveals two antiparallel beta sheets and an alpha helix stabilized by two Zn2+-binding clusters. The core secondary structures are similar to a rabphilin-3A Zn2+-binding Domain and to the C1 and LIM Domains. Phosphatidylinositol 3-phosphate binds to a pocket formed by the (R/K)(R/K)HHCR motif. A lattice contact shows how anionic ligands can interact with the phosphatidylinositol 3-phosphate-binding site. The tip of the FYVE Domain has basic and hydrophobic surfaces positioned so that nonspecific interactions with the phospholipid bilayer can abet specific binding to phosphatidylinositol 3-phosphate.
Anneclaude Gingras - One of the best experts on this subject based on the ideXlab platform.
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rag gtpases and phosphatidylinositol 3 phosphate mediate recruitment of the ap 5 spg11 spg15 complex
Journal of Cell Biology, 2021Co-Authors: Jennifer Hirst, G Hesketh, Anneclaude Gingras, Margaret S RobinsonAbstract:Adaptor protein complex 5 (AP-5) and its partners, SPG11 and SPG15, are recruited onto late endosomes and lysosomes. Here we show that recruitment of AP-5/SPG11/SPG15 is enhanced in starved cells and occurs by coincidence detection, requiring both phosphatidylinositol 3-phosphate (PI3P) and Rag GTPases. PI3P binding is via the SPG15 FYVE Domain, which, on its own, localizes to early endosomes. GDP-locked RagC promotes recruitment of AP-5/SPG11/SPG15, while GTP-locked RagA prevents its recruitment. Our results uncover an interplay between AP-5/SPG11/SPG15 and the mTORC1 pathway and help to explain the phenotype of AP-5/SPG11/SPG15 deficiency in patients, including the defect in autophagic lysosome reformation.
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Rag GTPases and phosphatidylinositol 3-phosphate mediate recruitment of the AP-5/SPG11/SPG15 complex.
The Journal of cell biology, 2021Co-Authors: Jennifer Hirst, Anneclaude Gingras, Geoffrey G Hesketh, Margaret S RobinsonAbstract:Adaptor protein complex 5 (AP-5) and its partners, SPG11 and SPG15, are recruited onto late endosomes and lysosomes. Here we show that recruitment of AP-5/SPG11/SPG15 is enhanced in starved cells and occurs by coincidence detection, requiring both phosphatidylinositol 3-phosphate (PI3P) and Rag GTPases. PI3P binding is via the SPG15 FYVE Domain, which, on its own, localizes to early endosomes. GDP-locked RagC promotes recruitment of AP-5/SPG11/SPG15, while GTP-locked RagA prevents its recruitment. Our results uncover an interplay between AP-5/SPG11/SPG15 and the mTORC1 pathway and help to explain the phenotype of AP-5/SPG11/SPG15 deficiency in patients, including the defect in autophagic lysosome reformation.
