PMID- 19901063
OWN - NLM
STAT- MEDLINE
DCOM- 20100121
LR  - 20211020
IS  - 1098-5522 (Electronic)
IS  - 0019-9567 (Print)
IS  - 0019-9567 (Linking)
VI  - 78
IP  - 1
DP  - 2010 Jan
TI  - Localization of phosphatidylinositol (3,4,5)-trisphosphate to phagosomes in 
      entamoeba histolytica achieved using glutathione S-transferase- and green 
      fluorescent protein-tagged lipid biosensors.
PG  - 125-37
LID - 10.1128/IAI.00719-09 [doi]
AB  - Entamoeba histolytica is an intestinal protozoan parasite that causes amoebic 
      dysentery and liver abscess. Phagocytosis by the parasite is a critical virulence 
      process, since it is a prerequisite for tissue invasion and establishment of 
      chronic infection. While the roles of many of the proteins that regulate 
      phagocytosis-related signaling events in E. histolytica have been characterized, 
      the functions of lipids in this cellular process remain largely unknown in this 
      parasite. In other systems, phosphatidylinositol (3,4,5)-trisphosphate (PIP(3)), 
      a major product of phosphoinositide 3 kinase (PI3-kinase) activity, is essential 
      for phagocytosis. Pleckstrin homology (PH) domains are protein domains that 
      specifically bind to PIP(3). In this study, we utilized glutathione S-transferase 
      (GST)- and green fluorescent protein (GFP)-labeled PH domains as lipid biosensors 
      to characterize the spatiotemporal aspects of PIP(3) distribution during various 
      endocytic processes in E. histolytica. PIP(3)-specific biosensors accumulated at 
      extending pseudopodia and in phagosomal cups in trophozoites exposed to 
      erythrocytes but did not localize to pinocytic compartments during the uptake of 
      a fluid-phase marker, dextran. Our results suggest that PIP(3) is involved in the 
      early stages of phagosome formation in E. histolytica. In addition, we 
      demonstrated that PIP(3) exists at high steady-state levels in the plasma 
      membrane of E. histolytica and that these levels, unlike those in mammalian 
      cells, are not abolished by serum withdrawal. Finally, expression of a PH domain 
      in trophozoites inhibited erythrophagocytosis and enhanced motility, providing 
      genetic evidence supporting the role of PI3-kinase signaling in these processes 
      in E. histolytica.
FAU - Byekova, Yevgeniya A
AU  - Byekova YA
AD  - Department of Biological Sciences, Clemson University, Clemson, South Carolina 
      29634, USA.
FAU - Powell, Rhonda R
AU  - Powell RR
FAU - Welter, Brenda H
AU  - Welter BH
FAU - Temesvari, Lesly A
AU  - Temesvari LA
LA  - eng
GR  - R01 AI046414/AI/NIAID NIH HHS/United States
GR  - R01 AI046414-08/AI/NIAID NIH HHS/United States
GR  - R01AI046414/AI/NIAID NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
PT  - Research Support, Non-U.S. Gov't
DEP - 20091109
PL  - United States
TA  - Infect Immun
JT  - Infection and immunity
JID - 0246127
RN  - 0 (Lipids)
RN  - 0 (Phosphatidylinositol Phosphates)
RN  - 0 (phosphatidylinositol 3,4,5-triphosphate)
RN  - 147336-22-9 (Green Fluorescent Proteins)
RN  - EC 2.5.1.18 (Glutathione Transferase)
RN  - EC 2.7.10.1 (Protein-Tyrosine Kinases)
RN  - EC 2.7.10.2 (Agammaglobulinaemia Tyrosine Kinase)
SB  - IM
MH  - Agammaglobulinaemia Tyrosine Kinase
MH  - Animals
MH  - Biosensing Techniques
MH  - Endocytosis/physiology
MH  - Entamoeba histolytica/cytology/*metabolism
MH  - Gene Expression Regulation/physiology
MH  - Glutathione Transferase/chemistry/*metabolism
MH  - Green Fluorescent Proteins/*chemistry/metabolism
MH  - Lipids/*chemistry
MH  - Phagosomes/*metabolism
MH  - Phosphatidylinositol Phosphates/*metabolism
MH  - Protein Structure, Tertiary
MH  - Protein-Tyrosine Kinases/chemistry
PMC - PMC2798214
EDAT- 2009/11/11 06:00
MHDA- 2010/01/22 06:00
PMCR- 2010/07/01
CRDT- 2009/11/11 06:00
PHST- 2009/11/11 06:00 [entrez]
PHST- 2009/11/11 06:00 [pubmed]
PHST- 2010/01/22 06:00 [medline]
PHST- 2010/07/01 00:00 [pmc-release]
AID - IAI.00719-09 [pii]
AID - 0719-09 [pii]
AID - 10.1128/IAI.00719-09 [doi]
PST - ppublish
SO  - Infect Immun. 2010 Jan;78(1):125-37. doi: 10.1128/IAI.00719-09. Epub 2009 Nov 9.