The asexual life cycle of Plasmodium falciparum parasites takes only 48 hours, allowing for rapid
replication. The continuous infection, rupturing and re-infection of erythrocytes results in the
pathogenicity of this disease. Schizogony (nuclear division) in P. falciparum parasites occurs via
alternation between the S and M phases of the cell cycle where DNA synthesis occurs in the
mature trophozoite and schizont stages, followed by mitosis to form daughter merozoites.
Merozoites then give rise to ring stages after they have infected erythrocytes and the ring stages
continue their development to trophozoites. This cyclic development, known as the intraerythrocytic
developmental cycle, has a unique transcriptional regulation, which is closely linked to
cell cycle regulation. However, the intricacies that these mechanisms are controlled by are still
unidentified. One of the means in which the P. falciparum parasite’s complex life cycle is controlled
is by means of epigenetics. Epigenetics refers to the heritable changes on a phenotypic level,
which are independent of changes on a genetic level.
One group of enzymes that participates in the parasite’s epigenetic control is the Plasmodium
histone deacetylases. Inhibition of histone deacetylases (HDACs) results in hyperacetylation,
which causes aberrant gene transcription and eventually results in parasite death. Comparative
analyses of three histone isolation methods and analysis of P. falciparum parasite histones and
their post-translational modifications (PTMs) by mass spectrometry techniques identified both
epigenetically relevant and novel PTMs in P. falciparum parasite histones and led to the discovery
of an adapted histone isolation method for investigation of histone PTM landscapes. When this
modified method was used for the investigation of histones that were isolated from P. falciparum
parasites treated with HDAC inhibitors compared to untreated parasites, differences were seen in the PTM landscape. Subsequent in silico screening strategies were used to identify ten compounds from the Medicines
for Malaria Venture (MMV) Malaria Box, which target the active site of the zinc-requiring PfHDAC1.
From these, eight compounds showed inhibition of proliferation of cultured P. falciparum parasites.
Ensuing, the adaptation of an HDAC assay to investigate histone deacetylase inhibition was used
to validate these compounds as possible PfHDAC1 inhibitors, with at least two of the compounds
showing significant inhibition of PfHDAC1 activity, comparable to that of the known HDAC inhibitor,
suberoylanilide hydroxamic acid, SAHA. The use of in silico screening of a large library of
compounds, such as the MMV Malaria Box, successfully narrows down candidates for possible
anti-malarials with drug-like properties by identification of their cellular targets. This work is
method-based and facilitates the investigation of the epigenetic landscape of histones, and the
identification of novel HDAC inhibitors. / Dissertation (MSc)--University of Pretoria, 2014. / gm2014 / Biochemistry / unrestricted
| Identifer | oai:union.ndltd.org:netd.ac.za/oai:union.ndltd.org:up/oai:repository.up.ac.za:2263/40345 |
| Date | January 2014 |
| Creators | Ismail, Nabila |
| Contributors | Birkholtz, Lyn-Marie, smile.nabila@gmail.com, Louw, A. I. |
| Source Sets | South African National ETD Portal |
| Language | English |
| Detected Language | English |
| Type | Dissertation |
| Rights | © 2013 University of Pretoria. All rights reserved. The copyright in this work vests in the University of Pretoria. No part of this work may be reproduced or transmitted in any form or by any means, without the prior written permission of the University of Pretoria. |
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