Polysome profiling of the malaria parasite Plasmodium falciparum

. Polysome profile from the malaria parasite Plasmodium falciparum, allowing for genome-wide analysis of translational regulation. [Display omitted] ► Intact polysomes were isolated from the malaria parasite Plasmodium falciparum. ► Polysome profiling offers a global view of translational activity i...

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Bibliographic Details
Published in:Molecular and biochemical parasitology 2011-09, Vol.179 (1), p.42-46
Main Authors: Lacsina, Joshua R., LaMonte, Gregory, Nicchitta, Christopher V., Chi, Jen-Tsan
Format: Article
Language:English
Subjects:
centrifugation
erythrocytes
Global
Malaria
messenger RNA
methodology
parasites
parasitology
Parasitology - methods
Plasmodium falciparum
Plasmodium falciparum - metabolism
polyribosomes
Polyribosomes - metabolism
Polysome
Post-transcriptional regulation
Protein Biosynthesis
RNA, Messenger - metabolism
sucrose
Translation
translation (genetics)
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Summary:. Polysome profile from the malaria parasite Plasmodium falciparum, allowing for genome-wide analysis of translational regulation. [Display omitted] ► Intact polysomes were isolated from the malaria parasite Plasmodium falciparum. ► Polysome profiling offers a global view of translational activity in the parasite. ► Allows determination of ribosomal loading and density for specific parasite mRNAs. In the malaria parasite Plasmodium falciparum, global studies of translational regulation have been hampered by the inability to isolate malaria polysomes. We describe here a novel method for polysome profiling in P. falciparum, a powerful approach which allows both a global view of translation and the measurement of ribosomal loading and density for specific mRNAs. Simultaneous lysis of infected erythrocytes and parasites releases stable, intact malaria polysomes, which are then purified by centrifugation through a sucrose cushion. The polysomes are resuspended, separated by velocity sedimentation and then fractionated, yielding a characteristic polysome profile reflecting the global level of translational activity in the parasite. RNA isolated from specific fractions can be used to determine the density of ribosomes loaded onto a particular transcript of interest, and is free of host ribosome contamination. Thus, our approach opens translational regulation in malaria to genome-wide analysis.
ISSN:0166-6851
1872-9428
DOI:10.1016/j.molbiopara.2011.05.003