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PMID: 11448993 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

The Toxoplasma gondii protein ROP2 mediates host organelle association with the parasitophorous vacuole membrane.

The Journal of cell biology ·Vol. 154 ·No. 1 ·2001-07-09 ·Pages 95-108

Sinai AP, Joiner KA

Abstract

Toxoplasma gondii replicates within a specialized vacuole surrounded by the parasitophorous vacuole membrane (PVM). The PVM forms intimate interactions with host mitochondria and endoplasmic reticulum (ER) in a process termed PVM-organelle association. In this study we identify a likely mediator of this process, the parasite protein ROP2. ROP2, which is localized to the PVM, is secreted from anterior organelles termed rhoptries during parasite invasion into host cells. The NH(2)-terminal domain of ROP2 (ROP2hc) within the PVM is exposed to the host cell cytosol, and has characteristics of a mitochondrial targeting signal. In in vitro assays, ROP2hc is partially translocated into the mitochondrial outer membrane and behaves like an integral membrane protein. Although ROP2hc does not translocate across the ER membrane, it does exhibit carbonate-resistant binding to this organelle. In vivo, ROP2hc expressed as a soluble fragment in the cytosol of uninfected cells associates with both mitochondria and ER. The 30-amino acid (aa) NH(2)-terminal sequence of ROP2hc, when fused to green fluorescent protein (GFP), is sufficient for mitochondrial targeting. Deletion of the 30-aa NH(2)-terminal signal from ROP2hc results in robust localization of the truncated protein to the ER. These results demonstrate a new mechanism for tight association of different membrane-bound organelles within the cell cytoplasm.

MeSH Terms
Amino Acid Sequence Animals CHO Cells Cells, Cultured Cricetinae Cytoplasm/metabolism Cytosol/metabolism Endoplasmic Reticulum/metabolism Fibroblasts/metabolism Green Fluorescent Proteins Humans Intracellular Membranes/metabolism Luminescent Proteins/metabolism Membrane Proteins/metabolism,physiology Mice Microsomes, Liver/metabolism Mitochondria/metabolism Mitochondria, Liver/metabolism Models, Genetic Molecular Sequence Data Nuclear Matrix/metabolism Plasmids/metabolism Protein Binding Protein Structure, Tertiary Protein Transport Protozoan Proteins/metabolism,physiology Subcellular Fractions Toxoplasma/chemistry Transfection Trypsin/pharmacology Vacuoles/metabolism
Chemicals
Luminescent Proteins Membrane Proteins Protozoan Proteins ROP 2 protein, Toxoplasma gondii Green Fluorescent Proteins Trypsin
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Sinai A P
Infectious Diseases Section, Department of Internal Medicine, Yale University School of Medicine, 333 Cedar Street, New Haven, CT 06520, USA. sinai@pop.uky.edu
Joiner K A
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Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.200101073
Email
sinai@pop.uky.edu
Published
2001-07-09
Pages
95-108
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2196872
Subset
IM
Grants
NIAID NIH HHS · N01AI30060 · United States
External Links
PubMed source DOI: 10.1083/jcb.200101073 PMC Full Text View journal details
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