Experimentally controlled downregulation of the histone chaperone FACT in Plasmodium berghei reveals that it is critical to male gamete fertility

Laurentino, Eliane C., Taylor, Sonya, Mair, Gunnar R., Lasonder, Edwin, Bartfai, Richard, Stunnenberg, Hendrik G., Kroeze, Hans, Ramesar, Jai, Franke‐Fayard, Blandine, Khan, Shahid M., Janse, Chris J. and Waters, Andrew P. (2011) Experimentally controlled downregulation of the histone chaperone FACT in Plasmodium berghei reveals that it is critical to male gamete fertility. Cellular Microbiology, 13 (12). pp. 1956-1974. ISSN 1462-5814

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Official URL: https://doi.org/10.1111/j.1462-5822.2011.01683.x


Human FACT (facilitates chromatin transcription) consists of the proteins SPT16 and SSRP1 and acts as a histone chaperone in the (dis)assembly of nucleosome (and thereby chromatin) structure during transcription and DNA replication. We identified a Plasmodium berghei protein, termed FACT-L, with homology to the SPT16 subunit of FACT. Epitope tagging of FACT-L showed nuclear localization with high expression in the nuclei of (activated) male gametocytes. The gene encoding FACT-L could not be deleted indicating an essential role during blood-stage development. Using a ‘promoter-swap’ approach whereby the fact-l promoter was replaced by an ‘asexual blood stage-specific’ promoter that is silent in gametocytes, transcription of fact-l in promoter-swap mutant gametocytes was downregulated compared with wild-type gametocytes. These mutant male gametocytes showed delayed DNA replication and gamete formation. Male gamete fertility was strongly reduced while female gamete fertility was unaffected; residual ookinetes generated oocysts that arrested early in development and failed to enter sporogony. Therefore FACT is critically involved in the formation of fertile male gametes and parasite transmission. ‘Promoter swapping’ is a powerful approach for the functional analysis of proteins in gametocytes (and beyond) that are essential during asexual blood-stage development.

Item Type: Article
Additional Information: Funding information: A postdoctoral fellowship from the Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) and from Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) to Eliane C. Laurentino is gratefully acknowledged. This work was supported by the Network of Excellence ‘Biology and Pathology of the Malaria Parasite’, BIOMALPAR of the European Commission (FP6), the Wellcome Trust and the Leverhulme Foundation. We thank Dr T. Pace and Dr I. Siden-Kiamos for communication of unpublished data and permission to cite it here. A.P.W., H.G.S and C.J.J. are supported by grants of the European Community's Seventh Framework Programme (FP7/2007-2013) under grant agreement N°242095.
Subjects: C700 Molecular Biology, Biophysics and Biochemistry
Department: Faculties > Health and Life Sciences > Applied Sciences
Depositing User: Elena Carlaw
Date Deposited: 08 Jun 2021 09:42
Last Modified: 31 Jul 2021 11:04
URI: http://nrl.northumbria.ac.uk/id/eprint/46377

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