Full Text Available

Access Repository
Note: Clicking the button above will open the full text document at the original institutional repository in a new window.
Cover Image

Expression and initial characterisation of the Plasmodium falciparum general transcription factors TFIIB and TLP

Malaria is a leading cause of morbidity and mortality worldwide, and results in approximately 600,000 deaths annually. The life cycle of the parasite is complex, and has several distinct stages of development. The transitions between these stages are brought about through tightly controlled and high...

Full description

Saved in:
Main Author: Bing, Steven
Other Authors: Oelgeschläger, Thomas
Format: Thesis
Language:English
Published: Department of Molecular and Cell Biology 2016
Subjects:
Molecular and Cell Biology
Tags: Add Tag
No Tags, Be the first to tag this record!
_version_ 1867613261660160000
access_status_str Open Access
author Bing, Steven
author2 Oelgeschläger, Thomas
author_browse Bing, Steven
Oelgeschläger, Thomas
author_facet Oelgeschläger, Thomas
Bing, Steven
author_sort Bing, Steven
collection Thesis
description Malaria is a leading cause of morbidity and mortality worldwide, and results in approximately 600,000 deaths annually. The life cycle of the parasite is complex, and has several distinct stages of development. The transitions between these stages are brought about through tightly controlled and highly synchronized changes in gene expression. Plasmodium falciparum causes the most lethal form of malaria in humans. The parasite is particularly virulent as it is able to evade immune detection by the infected host. This virulence is directly related to the expression of variable antigens on the surface of infected red blood cells. The control of gene expression is known to be largely regulated via RNA Polymerase II (RNAPII) transcription initiation, but in P. falciparum the underlying mechanisms have not been determined. This primarily because very little is known about both the key protein factors and DNA elements which guide the assembly of RNAPII components into the transcription initiation complex. Bioinformatics studies have shown that there is very little amino acid sequence conservation between human and Plasmodium RNAPII transcription initiation components. Together with the observation that the Plasmodium genome has an extremely high A+T content, this suggests that Plasmodium may have specific mechanisms to initiate transcription, which could be targeted by novel anti-malarials. The general transcription factor TFIIB and the TBP-like protein (TLP) are key proteins involved in the recognition of the core promoter, and the initiation of RNAPII transcription initiation complex assembly. TFIIB stabilises DNA binding of the primary promoter recognition factor, TATA-box binding protein (TBP), and is involved in promoter recognition through interactions with specific DNA sequences up- and downstream of the TBP DNA binding site. TBP-like protein is a member of the TBP protein family that has been implicated in life cycle stage specific gene transcription initiation in various eukaryotic model organisms. This research study reports the first successful expression and purification of recombinant epitope-tagged Plasmodium falciparum TFIIB and TLP proteins. Preliminary assays demonstrate DNA-binding activity for the recombinant Plasmodium TBP-like protein, and suggest DNA-binding activity in Plasmodium TFIIB protein, which has not been demonstrated before in eukaryotic TFIIB.
format Thesis
id oai:open.uct.ac.za:11427/19965
institution University of Cape Town (South Africa)
language eng
last_indexed 2026-06-10T12:33:19.547Z
license_str Not specified — see source repository
provenance_str_mv Harvested via OAI-PMH from UCTD — University of Cape Town Open Access Repository
publishDate 2016
publishDateRange 2016
publishDateSort 2016
publisher Department of Molecular and Cell Biology
publisherStr Department of Molecular and Cell Biology
record_format dspace
source_str UCTD — University of Cape Town Open Access Repository
spelling oai:open.uct.ac.za:11427/19965 Expression and initial characterisation of the Plasmodium falciparum general transcription factors TFIIB and TLP Bing, Steven Oelgeschläger, Thomas Molecular and Cell Biology Malaria is a leading cause of morbidity and mortality worldwide, and results in approximately 600,000 deaths annually. The life cycle of the parasite is complex, and has several distinct stages of development. The transitions between these stages are brought about through tightly controlled and highly synchronized changes in gene expression. Plasmodium falciparum causes the most lethal form of malaria in humans. The parasite is particularly virulent as it is able to evade immune detection by the infected host. This virulence is directly related to the expression of variable antigens on the surface of infected red blood cells. The control of gene expression is known to be largely regulated via RNA Polymerase II (RNAPII) transcription initiation, but in P. falciparum the underlying mechanisms have not been determined. This primarily because very little is known about both the key protein factors and DNA elements which guide the assembly of RNAPII components into the transcription initiation complex. Bioinformatics studies have shown that there is very little amino acid sequence conservation between human and Plasmodium RNAPII transcription initiation components. Together with the observation that the Plasmodium genome has an extremely high A+T content, this suggests that Plasmodium may have specific mechanisms to initiate transcription, which could be targeted by novel anti-malarials. The general transcription factor TFIIB and the TBP-like protein (TLP) are key proteins involved in the recognition of the core promoter, and the initiation of RNAPII transcription initiation complex assembly. TFIIB stabilises DNA binding of the primary promoter recognition factor, TATA-box binding protein (TBP), and is involved in promoter recognition through interactions with specific DNA sequences up- and downstream of the TBP DNA binding site. TBP-like protein is a member of the TBP protein family that has been implicated in life cycle stage specific gene transcription initiation in various eukaryotic model organisms. This research study reports the first successful expression and purification of recombinant epitope-tagged Plasmodium falciparum TFIIB and TLP proteins. Preliminary assays demonstrate DNA-binding activity for the recombinant Plasmodium TBP-like protein, and suggest DNA-binding activity in Plasmodium TFIIB protein, which has not been demonstrated before in eukaryotic TFIIB. 2016-06-09T11:17:38Z 2016-06-09T11:17:38Z 2015 Master Thesis Masters MSc http://hdl.handle.net/11427/19965 eng application/pdf Department of Molecular and Cell Biology Faculty of Science University of Cape Town
spellingShingle Molecular and Cell Biology
Bing, Steven
Expression and initial characterisation of the Plasmodium falciparum general transcription factors TFIIB and TLP
thesis_degree_str Master's
title Expression and initial characterisation of the Plasmodium falciparum general transcription factors TFIIB and TLP
title_full Expression and initial characterisation of the Plasmodium falciparum general transcription factors TFIIB and TLP
title_fullStr Expression and initial characterisation of the Plasmodium falciparum general transcription factors TFIIB and TLP
title_full_unstemmed Expression and initial characterisation of the Plasmodium falciparum general transcription factors TFIIB and TLP
title_short Expression and initial characterisation of the Plasmodium falciparum general transcription factors TFIIB and TLP
title_sort expression and initial characterisation of the plasmodium falciparum general transcription factors tfiib and tlp
topic Molecular and Cell Biology
url http://hdl.handle.net/11427/19965
work_keys_str_mv AT bingsteven expressionandinitialcharacterisationoftheplasmodiumfalciparumgeneraltranscriptionfactorstfiibandtlp

Similar Items