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

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 hom...

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Autores principales: 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, Waters, Andrew P
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Blackwell Publishing Ltd 2011
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3429858/
https://www.ncbi.nlm.nih.gov/pubmed/21899698
http://dx.doi.org/10.1111/j.1462-5822.2011.01683.x
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author 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
Waters, Andrew P
author_facet 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
Waters, Andrew P
author_sort Laurentino, Eliane C
collection PubMed
description 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.
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spelling pubmed-34298582012-08-29 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 Waters, Andrew P Cell Microbiol Original Articles 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. Blackwell Publishing Ltd 2011-12 /pmc/articles/PMC3429858/ /pubmed/21899698 http://dx.doi.org/10.1111/j.1462-5822.2011.01683.x Text en © 2011 Blackwell Publishing Ltd http://creativecommons.org/licenses/by/2.5/ Re-use of this article is permitted in accordance with the Creative Commons Deed, Attribution 2.5, which does not permit commercial exploitation.
spellingShingle Original Articles
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
Waters, Andrew P
Experimentally controlled downregulation of the histone chaperone FACT in Plasmodium berghei reveals that it is critical to male gamete fertility
title Experimentally controlled downregulation of the histone chaperone FACT in Plasmodium berghei reveals that it is critical to male gamete fertility
title_full Experimentally controlled downregulation of the histone chaperone FACT in Plasmodium berghei reveals that it is critical to male gamete fertility
title_fullStr Experimentally controlled downregulation of the histone chaperone FACT in Plasmodium berghei reveals that it is critical to male gamete fertility
title_full_unstemmed Experimentally controlled downregulation of the histone chaperone FACT in Plasmodium berghei reveals that it is critical to male gamete fertility
title_short Experimentally controlled downregulation of the histone chaperone FACT in Plasmodium berghei reveals that it is critical to male gamete fertility
title_sort experimentally controlled downregulation of the histone chaperone fact in plasmodium berghei reveals that it is critical to male gamete fertility
topic Original Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3429858/
https://www.ncbi.nlm.nih.gov/pubmed/21899698
http://dx.doi.org/10.1111/j.1462-5822.2011.01683.x
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