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Microtubular organization visualized by immunofluorescence microscopy during erythrocytic schizogony in Plasmodium falciparum and investigation of post-translational modifications of parasite tubulin

Published online by Cambridge University Press:  06 April 2009

M. Read ,
T. Sherwin ,
S. P. Holloway ,
K. Gull  and
J. E. Hyde
M. Read
Affiliation:
Department of Biochemistry and Applied Molecular Biology, University of Manchester Institute of Science and Technology (UMIST), Manchester M60 1QD, UK
T. Sherwin
Affiliation:
Department of Biochemistry and Molecular Biology, The Medical School, University of Manchester, Manchester M13 9PT, UK
S. P. Holloway
Affiliation:
Department of Biochemistry and Applied Molecular Biology, University of Manchester Institute of Science and Technology (UMIST), Manchester M60 1QD, UK
K. Gull
Affiliation:
Department of Biochemistry and Molecular Biology, The Medical School, University of Manchester, Manchester M13 9PT, UK
J. E. Hyde*
Affiliation:
Department of Biochemistry and Applied Molecular Biology, University of Manchester Institute of Science and Technology (UMIST), Manchester M60 1QD, UK
*
*Reprint requests to Dr J. E. Hyde, Department of Biochemistry and Applied Molecular Biology, University of Manchester Institute of Science and Technology (UMIST), P.O. Box 88, Manchester M60 1QD, UK.
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Summary

We describe a novel procedure for the immunofluorescent investigation of Plasmodium falciparum. This has allowed us to visualize clearly microtubular structures and their changing conformation through the erythrocytic cell-cycle, to the stage of cytodifferentiation leading to merozoite release. The images of spindle development we observed, together with an analysis of nuclear body numbers in large numbers of parasites, indicate that there is an apparent asynchrony in chromosomal multiplication within a single parasite. Using antibodies specific for post-translational modification of α- tubulin, we also demonstrate that the C-terminal tyrosine-containing epitope of P. falciparum α-tubulin I is similar to that of other organisms. Lysine-40 in the same molecule, a target for highly specific in vivo acetylation in some organisms, is unmodified in the blood stages we examined here. After in vitro acetylation of this residue, however, the epitope to which it contributes was recognized by antibody, showing that the conformation of this part of the molecule is also conserved, despite a lack of primary sequence homology immediately downstream of the target lysine residue.

Keywords

Plasmodium falciparumimmunofluorescencemicrotubulesmitosispost-translational modificationsacetylation.
Type
Research Article
Information
Parasitology , Volume 106 , Issue 3 , April 1993 , pp. 223 - 232
Copyright
Copyright © Cambridge University Press 1993

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