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Entamoeba histolytica: FYVE-finger domains, phosphatidylinositol 3-phosphate biosensors, associate with phagosomes but not fluid filled endosomes
Endocytosis is an important virulence function for Entamoeba histolytica, the causative agent of amoebic dysentery. Although a number of E. histolytica proteins that regulate this process have been identified, less is known about the role of lipids. In other systems, phosphatidylinositol 3-phosphate...
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| Published in: | Experimental parasitology 2006-04, Vol.112 (4), p.221-231 |
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| Main Authors: | , , , , , |
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| cites | cdi_FETCH-LOGICAL-c424t-b0ee5df0e61b471836269fd2eb2dcc55ef4ac76502bfca055710ac84de6c98103 |
| container_end_page | 231 |
| container_issue | 4 |
| container_start_page | 221 |
| container_title | Experimental parasitology |
| container_volume | 112 |
| creator | Powell, R.R. Welter, B.H. Hwu, R. Bowersox, B. Attaway, C. Temesvari, L.A. |
| description | Endocytosis is an important virulence function for
Entamoeba histolytica, the causative agent of amoebic dysentery. Although a number of
E. histolytica proteins that regulate this process have been identified, less is known about the role of lipids. In other systems, phosphatidylinositol 3-phosphate (PI3P), a product of phosphatidylinositol 3-kinase (PI 3-kinase), has been shown to be required for endocytosis. FYVE-finger domains are protein motifs that bind specifically to PI3P. Using a PI3P biosensor consisting of glutathione-
S-transferase (GST) fused to two tandem FYVE-finger domains, we have localized PI3P to phagosomes but not fluid-phase pinosomes in
E. histolytica, suggesting a role for PI3P in phagocytosis. Treatment of cells with PI 3-kinase inhibitors impaired GST-2×FYVE-phagosome association supporting the authenticity of the biosensor staining. However, treatment with PI 3-kinase inhibitors did not inhibit
E. histolytica-particle interaction, indicating that PI3P is not required for the initial step, but is required for subsequent steps of phagocytosis. |
| doi_str_mv | 10.1016/j.exppara.2005.11.013 |
| format | article |
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Entamoeba histolytica, the causative agent of amoebic dysentery. Although a number of
E. histolytica proteins that regulate this process have been identified, less is known about the role of lipids. In other systems, phosphatidylinositol 3-phosphate (PI3P), a product of phosphatidylinositol 3-kinase (PI 3-kinase), has been shown to be required for endocytosis. FYVE-finger domains are protein motifs that bind specifically to PI3P. Using a PI3P biosensor consisting of glutathione-
S-transferase (GST) fused to two tandem FYVE-finger domains, we have localized PI3P to phagosomes but not fluid-phase pinosomes in
E. histolytica, suggesting a role for PI3P in phagocytosis. Treatment of cells with PI 3-kinase inhibitors impaired GST-2×FYVE-phagosome association supporting the authenticity of the biosensor staining. However, treatment with PI 3-kinase inhibitors did not inhibit
E. histolytica-particle interaction, indicating that PI3P is not required for the initial step, but is required for subsequent steps of phagocytosis.</description><identifier>ISSN: 0014-4894</identifier><identifier>EISSN: 1090-2449</identifier><identifier>DOI: 10.1016/j.exppara.2005.11.013</identifier><identifier>PMID: 16387299</identifier><identifier>CODEN: EXPAAA</identifier><language>eng</language><publisher>San Diego, CA: Elsevier Inc</publisher><subject>Adhesion ; Amoeba ; Androstadienes - pharmacology ; Animals ; Biological and medical sciences ; Biosensing Techniques - methods ; Blotting, Western ; Chromones - pharmacology ; dimethylsulfoxide ; DMSO ; early endosome antigen 1 ; EEA1 ; EhPAK ; Electrophoresis, Polyacrylamide Gel ; Endocytosis ; Endosomes - physiology ; Entamoeba histolytica ; Entamoeba histolytica - physiology ; Enzyme Inhibitors - pharmacology ; Erythrocytes - immunology ; faciogenital dysplasia protein 1 ; FBS ; fetal bovine serum ; FGD1 ; Fundamental and applied biological sciences. Psychology ; FYVE-finger domain ; Gal/GalNAc ; galactose/ N-acetylgalactosamine ; GFP ; glutathione- S-transferase ; green fluorescent protein ; GST ; hepatocyte growth factor-regulated kinase substrate ; hRBCs ; Hrs ; human red blood cells ; Humans ; IPTG ; isopropyl-β- d-thiogalactopyranoside ; Life cycle. Host-agent relationship. Pathogenesis ; LY 294002 ; Morpholines - pharmacology ; p21-activated serine/threonine protein kinase ; PBS ; Phagocytosis ; Phagocytosis - drug effects ; Phagocytosis - physiology ; Phagosomes - physiology ; phosphate buffered saline ; Phosphatidylinositol 3-kinase ; Phosphatidylinositol 3-Kinases - antagonists & inhibitors ; Phosphatidylinositol 3-Kinases - physiology ; Phosphatidylinositol 3-phosphate ; phosphatidylinositol bisphosphate ; Phosphatidylinositol Phosphates - physiology ; phosphatidylinositol triphosphate ; PI 3-kinase ; PI3P ; Pinocytosis ; PIP 2 ; PIP 3 ; plekstrin homology ; Protozoa ; Rho guanine nucleotide exchange factor ; RhoGEF ; Silver Staining ; tetramethylrhodamine isothiocyanate ; TRITC ; Wortmannin</subject><ispartof>Experimental parasitology, 2006-04, Vol.112 (4), p.221-231</ispartof><rights>2005 Elsevier Inc.</rights><rights>2007 INIST-CNRS</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c424t-b0ee5df0e61b471836269fd2eb2dcc55ef4ac76502bfca055710ac84de6c98103</citedby><cites>FETCH-LOGICAL-c424t-b0ee5df0e61b471836269fd2eb2dcc55ef4ac76502bfca055710ac84de6c98103</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,780,784,27924,27925</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=18349584$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/16387299$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Powell, R.R.</creatorcontrib><creatorcontrib>Welter, B.H.</creatorcontrib><creatorcontrib>Hwu, R.</creatorcontrib><creatorcontrib>Bowersox, B.</creatorcontrib><creatorcontrib>Attaway, C.</creatorcontrib><creatorcontrib>Temesvari, L.A.</creatorcontrib><title>Entamoeba histolytica: FYVE-finger domains, phosphatidylinositol 3-phosphate biosensors, associate with phagosomes but not fluid filled endosomes</title><title>Experimental parasitology</title><addtitle>Exp Parasitol</addtitle><description>Endocytosis is an important virulence function for
Entamoeba histolytica, the causative agent of amoebic dysentery. Although a number of
E. histolytica proteins that regulate this process have been identified, less is known about the role of lipids. In other systems, phosphatidylinositol 3-phosphate (PI3P), a product of phosphatidylinositol 3-kinase (PI 3-kinase), has been shown to be required for endocytosis. FYVE-finger domains are protein motifs that bind specifically to PI3P. Using a PI3P biosensor consisting of glutathione-
S-transferase (GST) fused to two tandem FYVE-finger domains, we have localized PI3P to phagosomes but not fluid-phase pinosomes in
E. histolytica, suggesting a role for PI3P in phagocytosis. Treatment of cells with PI 3-kinase inhibitors impaired GST-2×FYVE-phagosome association supporting the authenticity of the biosensor staining. However, treatment with PI 3-kinase inhibitors did not inhibit
E. histolytica-particle interaction, indicating that PI3P is not required for the initial step, but is required for subsequent steps of phagocytosis.</description><subject>Adhesion</subject><subject>Amoeba</subject><subject>Androstadienes - pharmacology</subject><subject>Animals</subject><subject>Biological and medical sciences</subject><subject>Biosensing Techniques - methods</subject><subject>Blotting, Western</subject><subject>Chromones - pharmacology</subject><subject>dimethylsulfoxide</subject><subject>DMSO</subject><subject>early endosome antigen 1</subject><subject>EEA1</subject><subject>EhPAK</subject><subject>Electrophoresis, Polyacrylamide Gel</subject><subject>Endocytosis</subject><subject>Endosomes - physiology</subject><subject>Entamoeba histolytica</subject><subject>Entamoeba histolytica - physiology</subject><subject>Enzyme Inhibitors - pharmacology</subject><subject>Erythrocytes - immunology</subject><subject>faciogenital dysplasia protein 1</subject><subject>FBS</subject><subject>fetal bovine serum</subject><subject>FGD1</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>FYVE-finger domain</subject><subject>Gal/GalNAc</subject><subject>galactose/ N-acetylgalactosamine</subject><subject>GFP</subject><subject>glutathione- S-transferase</subject><subject>green fluorescent protein</subject><subject>GST</subject><subject>hepatocyte growth factor-regulated kinase substrate</subject><subject>hRBCs</subject><subject>Hrs</subject><subject>human red blood cells</subject><subject>Humans</subject><subject>IPTG</subject><subject>isopropyl-β- d-thiogalactopyranoside</subject><subject>Life cycle. Host-agent relationship. Pathogenesis</subject><subject>LY 294002</subject><subject>Morpholines - pharmacology</subject><subject>p21-activated serine/threonine protein kinase</subject><subject>PBS</subject><subject>Phagocytosis</subject><subject>Phagocytosis - drug effects</subject><subject>Phagocytosis - physiology</subject><subject>Phagosomes - physiology</subject><subject>phosphate buffered saline</subject><subject>Phosphatidylinositol 3-kinase</subject><subject>Phosphatidylinositol 3-Kinases - antagonists & inhibitors</subject><subject>Phosphatidylinositol 3-Kinases - physiology</subject><subject>Phosphatidylinositol 3-phosphate</subject><subject>phosphatidylinositol bisphosphate</subject><subject>Phosphatidylinositol Phosphates - physiology</subject><subject>phosphatidylinositol triphosphate</subject><subject>PI 3-kinase</subject><subject>PI3P</subject><subject>Pinocytosis</subject><subject>PIP 2</subject><subject>PIP 3</subject><subject>plekstrin homology</subject><subject>Protozoa</subject><subject>Rho guanine nucleotide exchange factor</subject><subject>RhoGEF</subject><subject>Silver Staining</subject><subject>tetramethylrhodamine isothiocyanate</subject><subject>TRITC</subject><subject>Wortmannin</subject><issn>0014-4894</issn><issn>1090-2449</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2006</creationdate><recordtype>article</recordtype><recordid>eNqF0c1u1DAUBeAIgehQeASQN7Aiw7Xj_JhNhaopIFViA0isLMe-6XiU2ME3AeYxeGMymqAuu7J0_R3b8smylxy2HHj17rDFP-NoktkKgHLL-RZ48SjbcFCQCynV42wDwGUuGyUvsmdEBwBouJBPswteFU0tlNpkf3dhMkPE1rC9pyn2x8lb857d_Pi-yzsf7jAxFwfjA71l4z7SuDeTd8feh0h-8azI_4-RtT4SBoppwYYoWn-a_vbTfsmau0hxQGLtPLEQJ9b1s3es832PjmFw5-3n2ZPO9IQv1vUy-3az-3r9Kb_98vHz9Yfb3Eohp7wFxNJ1gBVvZc2bohKV6pzAVjhryxI7aWxdlSDazhooy5qDsY10WFnVcCguszfnc8cUf85Ikx48Wex7EzDOpKu6rqAq1YNQgFKirvkCyzO0KRIl7PSY_GDSUXPQp9L0Qa-l6VNpmnO9lLbkXq0XzO2A7j61trSA1yswZE3fJROsp3vXFFKVjVzc1dnh8m-_PCZN1mOw6HxCO2kX_QNP-QfZr7v2</recordid><startdate>20060401</startdate><enddate>20060401</enddate><creator>Powell, R.R.</creator><creator>Welter, B.H.</creator><creator>Hwu, R.</creator><creator>Bowersox, B.</creator><creator>Attaway, C.</creator><creator>Temesvari, L.A.</creator><general>Elsevier Inc</general><general>Elsevier</general><scope>IQODW</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7QO</scope><scope>8FD</scope><scope>FR3</scope><scope>M7N</scope><scope>P64</scope><scope>7X8</scope></search><sort><creationdate>20060401</creationdate><title>Entamoeba histolytica: FYVE-finger domains, phosphatidylinositol 3-phosphate biosensors, associate with phagosomes but not fluid filled endosomes</title><author>Powell, R.R. ; Welter, B.H. ; Hwu, R. ; Bowersox, B. ; Attaway, C. ; Temesvari, L.A.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c424t-b0ee5df0e61b471836269fd2eb2dcc55ef4ac76502bfca055710ac84de6c98103</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2006</creationdate><topic>Adhesion</topic><topic>Amoeba</topic><topic>Androstadienes - pharmacology</topic><topic>Animals</topic><topic>Biological and medical sciences</topic><topic>Biosensing Techniques - methods</topic><topic>Blotting, Western</topic><topic>Chromones - pharmacology</topic><topic>dimethylsulfoxide</topic><topic>DMSO</topic><topic>early endosome antigen 1</topic><topic>EEA1</topic><topic>EhPAK</topic><topic>Electrophoresis, Polyacrylamide Gel</topic><topic>Endocytosis</topic><topic>Endosomes - physiology</topic><topic>Entamoeba histolytica</topic><topic>Entamoeba histolytica - physiology</topic><topic>Enzyme Inhibitors - pharmacology</topic><topic>Erythrocytes - immunology</topic><topic>faciogenital dysplasia protein 1</topic><topic>FBS</topic><topic>fetal bovine serum</topic><topic>FGD1</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>FYVE-finger domain</topic><topic>Gal/GalNAc</topic><topic>galactose/ N-acetylgalactosamine</topic><topic>GFP</topic><topic>glutathione- S-transferase</topic><topic>green fluorescent protein</topic><topic>GST</topic><topic>hepatocyte growth factor-regulated kinase substrate</topic><topic>hRBCs</topic><topic>Hrs</topic><topic>human red blood cells</topic><topic>Humans</topic><topic>IPTG</topic><topic>isopropyl-β- d-thiogalactopyranoside</topic><topic>Life cycle. Host-agent relationship. Pathogenesis</topic><topic>LY 294002</topic><topic>Morpholines - pharmacology</topic><topic>p21-activated serine/threonine protein kinase</topic><topic>PBS</topic><topic>Phagocytosis</topic><topic>Phagocytosis - drug effects</topic><topic>Phagocytosis - physiology</topic><topic>Phagosomes - physiology</topic><topic>phosphate buffered saline</topic><topic>Phosphatidylinositol 3-kinase</topic><topic>Phosphatidylinositol 3-Kinases - antagonists & inhibitors</topic><topic>Phosphatidylinositol 3-Kinases - physiology</topic><topic>Phosphatidylinositol 3-phosphate</topic><topic>phosphatidylinositol bisphosphate</topic><topic>Phosphatidylinositol Phosphates - physiology</topic><topic>phosphatidylinositol triphosphate</topic><topic>PI 3-kinase</topic><topic>PI3P</topic><topic>Pinocytosis</topic><topic>PIP 2</topic><topic>PIP 3</topic><topic>plekstrin homology</topic><topic>Protozoa</topic><topic>Rho guanine nucleotide exchange factor</topic><topic>RhoGEF</topic><topic>Silver Staining</topic><topic>tetramethylrhodamine isothiocyanate</topic><topic>TRITC</topic><topic>Wortmannin</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Powell, R.R.</creatorcontrib><creatorcontrib>Welter, B.H.</creatorcontrib><creatorcontrib>Hwu, R.</creatorcontrib><creatorcontrib>Bowersox, B.</creatorcontrib><creatorcontrib>Attaway, C.</creatorcontrib><creatorcontrib>Temesvari, L.A.</creatorcontrib><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Biotechnology Research Abstracts</collection><collection>Technology Research Database</collection><collection>Engineering Research Database</collection><collection>Algology Mycology and Protozoology Abstracts (Microbiology C)</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>Experimental parasitology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Powell, R.R.</au><au>Welter, B.H.</au><au>Hwu, R.</au><au>Bowersox, B.</au><au>Attaway, C.</au><au>Temesvari, L.A.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Entamoeba histolytica: FYVE-finger domains, phosphatidylinositol 3-phosphate biosensors, associate with phagosomes but not fluid filled endosomes</atitle><jtitle>Experimental parasitology</jtitle><addtitle>Exp Parasitol</addtitle><date>2006-04-01</date><risdate>2006</risdate><volume>112</volume><issue>4</issue><spage>221</spage><epage>231</epage><pages>221-231</pages><issn>0014-4894</issn><eissn>1090-2449</eissn><coden>EXPAAA</coden><abstract>Endocytosis is an important virulence function for
Entamoeba histolytica, the causative agent of amoebic dysentery. Although a number of
E. histolytica proteins that regulate this process have been identified, less is known about the role of lipids. In other systems, phosphatidylinositol 3-phosphate (PI3P), a product of phosphatidylinositol 3-kinase (PI 3-kinase), has been shown to be required for endocytosis. FYVE-finger domains are protein motifs that bind specifically to PI3P. Using a PI3P biosensor consisting of glutathione-
S-transferase (GST) fused to two tandem FYVE-finger domains, we have localized PI3P to phagosomes but not fluid-phase pinosomes in
E. histolytica, suggesting a role for PI3P in phagocytosis. Treatment of cells with PI 3-kinase inhibitors impaired GST-2×FYVE-phagosome association supporting the authenticity of the biosensor staining. However, treatment with PI 3-kinase inhibitors did not inhibit
E. histolytica-particle interaction, indicating that PI3P is not required for the initial step, but is required for subsequent steps of phagocytosis.</abstract><cop>San Diego, CA</cop><pub>Elsevier Inc</pub><pmid>16387299</pmid><doi>10.1016/j.exppara.2005.11.013</doi><tpages>11</tpages></addata></record> |
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| ispartof | Experimental parasitology, 2006-04, Vol.112 (4), p.221-231 |
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| subjects | Adhesion Amoeba Androstadienes - pharmacology Animals Biological and medical sciences Biosensing Techniques - methods Blotting, Western Chromones - pharmacology dimethylsulfoxide DMSO early endosome antigen 1 EEA1 EhPAK Electrophoresis, Polyacrylamide Gel Endocytosis Endosomes - physiology Entamoeba histolytica Entamoeba histolytica - physiology Enzyme Inhibitors - pharmacology Erythrocytes - immunology faciogenital dysplasia protein 1 FBS fetal bovine serum FGD1 Fundamental and applied biological sciences. Psychology FYVE-finger domain Gal/GalNAc galactose/ N-acetylgalactosamine GFP glutathione- S-transferase green fluorescent protein GST hepatocyte growth factor-regulated kinase substrate hRBCs Hrs human red blood cells Humans IPTG isopropyl-β- d-thiogalactopyranoside Life cycle. Host-agent relationship. Pathogenesis LY 294002 Morpholines - pharmacology p21-activated serine/threonine protein kinase PBS Phagocytosis Phagocytosis - drug effects Phagocytosis - physiology Phagosomes - physiology phosphate buffered saline Phosphatidylinositol 3-kinase Phosphatidylinositol 3-Kinases - antagonists & inhibitors Phosphatidylinositol 3-Kinases - physiology Phosphatidylinositol 3-phosphate phosphatidylinositol bisphosphate Phosphatidylinositol Phosphates - physiology phosphatidylinositol triphosphate PI 3-kinase PI3P Pinocytosis PIP 2 PIP 3 plekstrin homology Protozoa Rho guanine nucleotide exchange factor RhoGEF Silver Staining tetramethylrhodamine isothiocyanate TRITC Wortmannin |
| title | Entamoeba histolytica: FYVE-finger domains, phosphatidylinositol 3-phosphate biosensors, associate with phagosomes but not fluid filled endosomes |
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