Inducing controlled cell cycle arrest and re-entry during asexual proliferation of Plasmodium falciparum malaria parasites

Riëtte van Biljon, Jandeli Niemand, Roelof van Wyk, Katherine Clark, Bianca Verlinden, Clarissa Abrie, Hilde von Grüning, Werner Smidt, Annél Smit, Janette Reader, Heather Painter, Manuel Llinás, Christian Doerig, Lyn Marié Birkholtz

Research output: Contribution to journalArticleResearchpeer-review

Abstract

The life cycle of the malaria parasite Plasmodium falciparum is tightly regulated, oscillating between stages of intense proliferation and quiescence. Cyclic 48-hour asexual replication of Plasmodium is markedly different from cell division in higher eukaryotes, and mechanistically poorly understood. Here, we report tight synchronisation of malaria parasites during the early phases of the cell cycle by exposure to DL-α-difluoromethylornithine (DFMO), which results in the depletion of polyamines. This induces an inescapable cell cycle arrest in G1 (~15 hours post-invasion) by blocking G1/S transition. Cell cycle-arrested parasites enter a quiescent G0-like state but, upon addition of exogenous polyamines, re-initiate their cell cycle. This ability to halt malaria parasites at a specific point in their cell cycle, and to subsequently trigger re-entry into the cell cycle, provides a valuable framework to investigate cell cycle regulation in these parasites. We subsequently used gene expression analyses to show that re-entry into the cell cycle involves expression of Ca2+-sensitive (cdpk4 and pk2) and mitotic kinases (nima and ark2), with deregulation of the pre-replicative complex associated with expression of pk2. Changes in gene expression could be driven through transcription factors MYB1 and two ApiAP2 family members. This new approach to parasite synchronisation therefore expands our currently limited toolkit to investigate cell cycle regulation in malaria parasites.

Original languageEnglish
Article number16581
JournalScientific Reports
Volume8
Issue number1
DOIs
Publication statusPublished - 1 Dec 2018

Cite this

van Biljon, R., Niemand, J., van Wyk, R., Clark, K., Verlinden, B., Abrie, C., ... Birkholtz, L. M. (2018). Inducing controlled cell cycle arrest and re-entry during asexual proliferation of Plasmodium falciparum malaria parasites. Scientific Reports, 8(1), [16581]. https://doi.org/10.1038/s41598-018-34964-w
van Biljon, Riëtte ; Niemand, Jandeli ; van Wyk, Roelof ; Clark, Katherine ; Verlinden, Bianca ; Abrie, Clarissa ; von Grüning, Hilde ; Smidt, Werner ; Smit, Annél ; Reader, Janette ; Painter, Heather ; Llinás, Manuel ; Doerig, Christian ; Birkholtz, Lyn Marié. / Inducing controlled cell cycle arrest and re-entry during asexual proliferation of Plasmodium falciparum malaria parasites. In: Scientific Reports. 2018 ; Vol. 8, No. 1.
@article{3a4d99b59e1b48d0bfb58157d2944e3e,
title = "Inducing controlled cell cycle arrest and re-entry during asexual proliferation of Plasmodium falciparum malaria parasites",
abstract = "The life cycle of the malaria parasite Plasmodium falciparum is tightly regulated, oscillating between stages of intense proliferation and quiescence. Cyclic 48-hour asexual replication of Plasmodium is markedly different from cell division in higher eukaryotes, and mechanistically poorly understood. Here, we report tight synchronisation of malaria parasites during the early phases of the cell cycle by exposure to DL-α-difluoromethylornithine (DFMO), which results in the depletion of polyamines. This induces an inescapable cell cycle arrest in G1 (~15 hours post-invasion) by blocking G1/S transition. Cell cycle-arrested parasites enter a quiescent G0-like state but, upon addition of exogenous polyamines, re-initiate their cell cycle. This ability to halt malaria parasites at a specific point in their cell cycle, and to subsequently trigger re-entry into the cell cycle, provides a valuable framework to investigate cell cycle regulation in these parasites. We subsequently used gene expression analyses to show that re-entry into the cell cycle involves expression of Ca2+-sensitive (cdpk4 and pk2) and mitotic kinases (nima and ark2), with deregulation of the pre-replicative complex associated with expression of pk2. Changes in gene expression could be driven through transcription factors MYB1 and two ApiAP2 family members. This new approach to parasite synchronisation therefore expands our currently limited toolkit to investigate cell cycle regulation in malaria parasites.",
author = "{van Biljon}, Ri{\"e}tte and Jandeli Niemand and {van Wyk}, Roelof and Katherine Clark and Bianca Verlinden and Clarissa Abrie and {von Gr{\"u}ning}, Hilde and Werner Smidt and Ann{\'e}l Smit and Janette Reader and Heather Painter and Manuel Llin{\'a}s and Christian Doerig and Birkholtz, {Lyn Mari{\'e}}",
year = "2018",
month = "12",
day = "1",
doi = "10.1038/s41598-018-34964-w",
language = "English",
volume = "8",
journal = "Scientific Reports",
issn = "2045-2322",
publisher = "Nature Publishing Group",
number = "1",

}

van Biljon, R, Niemand, J, van Wyk, R, Clark, K, Verlinden, B, Abrie, C, von Grüning, H, Smidt, W, Smit, A, Reader, J, Painter, H, Llinás, M, Doerig, C & Birkholtz, LM 2018, 'Inducing controlled cell cycle arrest and re-entry during asexual proliferation of Plasmodium falciparum malaria parasites' Scientific Reports, vol. 8, no. 1, 16581. https://doi.org/10.1038/s41598-018-34964-w

Inducing controlled cell cycle arrest and re-entry during asexual proliferation of Plasmodium falciparum malaria parasites. / van Biljon, Riëtte; Niemand, Jandeli; van Wyk, Roelof; Clark, Katherine; Verlinden, Bianca; Abrie, Clarissa; von Grüning, Hilde; Smidt, Werner; Smit, Annél; Reader, Janette; Painter, Heather; Llinás, Manuel; Doerig, Christian; Birkholtz, Lyn Marié.

In: Scientific Reports, Vol. 8, No. 1, 16581, 01.12.2018.

Research output: Contribution to journalArticleResearchpeer-review

TY - JOUR

T1 - Inducing controlled cell cycle arrest and re-entry during asexual proliferation of Plasmodium falciparum malaria parasites

AU - van Biljon, Riëtte

AU - Niemand, Jandeli

AU - van Wyk, Roelof

AU - Clark, Katherine

AU - Verlinden, Bianca

AU - Abrie, Clarissa

AU - von Grüning, Hilde

AU - Smidt, Werner

AU - Smit, Annél

AU - Reader, Janette

AU - Painter, Heather

AU - Llinás, Manuel

AU - Doerig, Christian

AU - Birkholtz, Lyn Marié

PY - 2018/12/1

Y1 - 2018/12/1

N2 - The life cycle of the malaria parasite Plasmodium falciparum is tightly regulated, oscillating between stages of intense proliferation and quiescence. Cyclic 48-hour asexual replication of Plasmodium is markedly different from cell division in higher eukaryotes, and mechanistically poorly understood. Here, we report tight synchronisation of malaria parasites during the early phases of the cell cycle by exposure to DL-α-difluoromethylornithine (DFMO), which results in the depletion of polyamines. This induces an inescapable cell cycle arrest in G1 (~15 hours post-invasion) by blocking G1/S transition. Cell cycle-arrested parasites enter a quiescent G0-like state but, upon addition of exogenous polyamines, re-initiate their cell cycle. This ability to halt malaria parasites at a specific point in their cell cycle, and to subsequently trigger re-entry into the cell cycle, provides a valuable framework to investigate cell cycle regulation in these parasites. We subsequently used gene expression analyses to show that re-entry into the cell cycle involves expression of Ca2+-sensitive (cdpk4 and pk2) and mitotic kinases (nima and ark2), with deregulation of the pre-replicative complex associated with expression of pk2. Changes in gene expression could be driven through transcription factors MYB1 and two ApiAP2 family members. This new approach to parasite synchronisation therefore expands our currently limited toolkit to investigate cell cycle regulation in malaria parasites.

AB - The life cycle of the malaria parasite Plasmodium falciparum is tightly regulated, oscillating between stages of intense proliferation and quiescence. Cyclic 48-hour asexual replication of Plasmodium is markedly different from cell division in higher eukaryotes, and mechanistically poorly understood. Here, we report tight synchronisation of malaria parasites during the early phases of the cell cycle by exposure to DL-α-difluoromethylornithine (DFMO), which results in the depletion of polyamines. This induces an inescapable cell cycle arrest in G1 (~15 hours post-invasion) by blocking G1/S transition. Cell cycle-arrested parasites enter a quiescent G0-like state but, upon addition of exogenous polyamines, re-initiate their cell cycle. This ability to halt malaria parasites at a specific point in their cell cycle, and to subsequently trigger re-entry into the cell cycle, provides a valuable framework to investigate cell cycle regulation in these parasites. We subsequently used gene expression analyses to show that re-entry into the cell cycle involves expression of Ca2+-sensitive (cdpk4 and pk2) and mitotic kinases (nima and ark2), with deregulation of the pre-replicative complex associated with expression of pk2. Changes in gene expression could be driven through transcription factors MYB1 and two ApiAP2 family members. This new approach to parasite synchronisation therefore expands our currently limited toolkit to investigate cell cycle regulation in malaria parasites.

UR - http://www.scopus.com/inward/record.url?scp=85056266415&partnerID=8YFLogxK

U2 - 10.1038/s41598-018-34964-w

DO - 10.1038/s41598-018-34964-w

M3 - Article

VL - 8

JO - Scientific Reports

JF - Scientific Reports

SN - 2045-2322

IS - 1

M1 - 16581

ER -