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Functional analysis of the leading malaria vaccine candidate AMA-1 reveals an essential role for the cytoplasmic domain in the invasion process

Treeck, Moritz, Zacherl, Sonja, Herrmann, Susann, Cabrera, Ana, Kono, Maya, Struck, Nicole S., Engelberg, Klemens, Haase, Silvia, Frischknecht, Friedrich, Miura, Kota, Spielmann, Tobias and Gilberger, Tim W. 2009, Functional analysis of the leading malaria vaccine candidate AMA-1 reveals an essential role for the cytoplasmic domain in the invasion process, PLoS pathogens, vol. 5, no. 3, pp. 1-13, doi: 10.1371/journal.ppat.1000322.

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Title Functional analysis of the leading malaria vaccine candidate AMA-1 reveals an essential role for the cytoplasmic domain in the invasion process
Author(s) Treeck, Moritz
Zacherl, Sonja
Herrmann, Susann
Cabrera, Ana
Kono, Maya
Struck, Nicole S.
Engelberg, Klemens
Haase, Silvia
Frischknecht, Friedrich
Miura, Kota
Spielmann, Tobias
Gilberger, Tim W.
Journal name PLoS pathogens
Volume number 5
Issue number 3
Article ID e1000322
Start page 1
End page 13
Publisher Public Library of Science
Place of publication San Francisco, Calif.
Publication date 2009-03
ISSN 1553-7366
1553-7374
Keyword(s) malaria
Summary A key process in the lifecycle of the malaria parasite Plasmodium falciparum is the fast invasion of human erythrocytes. Entry into the host cell requires the apical membrane antigen 1 (AMA-1), a type I transmembrane protein located in the micronemes of the merozoite. Although AMA-1 is evolving into the leading blood-stage malaria vaccine candidate, its precise role in invasion is still unclear. We investigate AMA-1 function using live video microscopy in the absence and presence of an AMA-1 inhibitory peptide. This data reveals a crucial function of AMA-1 during the primary contact period upstream of the entry process at around the time of moving junction formation. We generate a Plasmodium falciparum cell line that expresses a functional GFP-tagged AMA-1. This allows the visualization of the dynamics of AMA-1 in live parasites. We functionally validate the ectopically expressed AMA-1 by establishing a complementation assay based on strain-specific inhibition. This method provides the basis for the functional analysis of essential genes that are refractory to any genetic manipulation. Using the complementation assay, we show that the cytoplasmic domain of AMA-1 is not required for correct trafficking and surface translocation but is essential for AMA-1 function. Although this function can be mimicked by the highly conserved cytoplasmic domains of P. vivax and P. berghei, the exchange with the heterologous domain of the microneme protein EBA-175 or the rhoptry protein Rh2b leads to a loss of function. We identify several residues in the cytoplasmic tail that are essential for AMA-1 function. We validate this data using additional transgenic parasite lines expressing AMA-1 mutants with TY1 epitopes. We show that the cytoplasmic domain of AMA-1 is phosphorylated. Mutational analysis suggests an important role for the phosphorylation in the invasion process, which might translate into novel therapeutic strategies.
Language eng
DOI 10.1371/journal.ppat.1000322
Field of Research 110803 Medical Parasitology
Socio Economic Objective 970103 Expanding Knowledge in the Chemical Sciences
HERDC Research category C1.1 Refereed article in a scholarly journal
Copyright notice ©2009, The Authors
Free to Read? Yes
Use Rights Creative Commons Attribution licence
Persistent URL http://hdl.handle.net/10536/DRO/DU:30040965

Document type: Journal Article
Collections: School of Medicine
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Created: Wed, 07 Dec 2011, 11:14:52 EST by Leanne Swaneveld

Every reasonable effort has been made to ensure that permission has been obtained for items included in DRO. If you believe that your rights have been infringed by this repository, please contact drosupport@deakin.edu.au.