Abstract
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.
DOI
10.1111/j.1462-5822.2011.01683.x
Publication Date
2011-12-01
Publication Title
Cellular Microbiology
Volume
13
Issue
12
Publisher
Hindawi Limited
ISSN
1462-5822
Embargo Period
2024-11-19
First Page
1956
Last Page
1974
Recommended Citation
Laurentino, E., Taylor, S., Mair, G., Lasonder, E., & et al. (2011) 'Experimentally controlled downregulation of the histone chaperone FACT inPlasmodium bergheireveals that it is critical to male gamete fertility', Cellular Microbiology, 13(12), pp. 1956-1974. Hindawi Limited: Available at: https://doi.org/10.1111/j.1462-5822.2011.01683.x
