Protein Kinase A Is Essential for Invasion of <named-content content-type="genus-species">Plasmodium falciparum</named-content> into Human Erythrocytes

ABSTRACT Understanding the mechanisms behind host cell invasion by Plasmodium falciparum remains a major hurdle to developing antimalarial therapeutics that target the asexual cycle and the symptomatic stage of malaria. Host cell entry is enabled by a multitude of precisely timed and tightly regulat...

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Autores principales: Mary-Louise Wilde, Tony Triglia, Danushka Marapana, Jennifer K. Thompson, Alexei A. Kouzmitchev, Hayley E. Bullen, Paul R. Gilson, Alan F. Cowman, Christopher J. Tonkin
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Publicado: American Society for Microbiology 2019
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spelling oai:doaj.org-article:e57a13863f35463ca89cb2fe9e8b1b582021-11-15T15:59:41ZProtein Kinase A Is Essential for Invasion of <named-content content-type="genus-species">Plasmodium falciparum</named-content> into Human Erythrocytes10.1128/mBio.01972-192150-7511https://doaj.org/article/e57a13863f35463ca89cb2fe9e8b1b582019-10-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.01972-19https://doaj.org/toc/2150-7511ABSTRACT Understanding the mechanisms behind host cell invasion by Plasmodium falciparum remains a major hurdle to developing antimalarial therapeutics that target the asexual cycle and the symptomatic stage of malaria. Host cell entry is enabled by a multitude of precisely timed and tightly regulated receptor-ligand interactions. Cyclic nucleotide signaling has been implicated in regulating parasite invasion, and an important downstream effector of the cAMP-signaling pathway is protein kinase A (PKA), a cAMP-dependent protein kinase. There is increasing evidence that P. falciparum PKA (PfPKA) is responsible for phosphorylation of the cytoplasmic domain of P. falciparum apical membrane antigen 1 (PfAMA1) at Ser610, a cAMP-dependent event that is crucial for successful parasite invasion. In the present study, CRISPR-Cas9 and conditional gene deletion (dimerizable cre) technologies were implemented to generate a P. falciparum parasite line in which expression of the catalytic subunit of PfPKA (PfPKAc) is under conditional control, demonstrating highly efficient dimerizable Cre recombinase (DiCre)-mediated gene excision and complete knockdown of protein expression. Parasites lacking PfPKAc show severely reduced growth after one intraerythrocytic growth cycle and are deficient in host cell invasion, as highlighted by live-imaging experiments. Furthermore, PfPKAc-deficient parasites are unable to phosphorylate PfAMA1 at Ser610. This work not only identifies an essential role for PfPKAc in the P. falciparum asexual life cycle but also confirms that PfPKAc is the kinase responsible for phosphorylating PfAMA1 Ser610. IMPORTANCE Malaria continues to present a major global health burden, particularly in low-resource countries. Plasmodium falciparum, the parasite responsible for the most severe form of malaria, causes disease through rapid and repeated rounds of invasion and replication within red blood cells. Invasion into red blood cells is essential for P. falciparum survival, and the molecular events mediating this process have gained much attention as potential therapeutic targets. With no effective vaccine available, and with the emergence of resistance to antimalarials, there is an urgent need for the development of new therapeutics. Our research has used genetic techniques to provide evidence of an essential protein kinase involved in P. falciparum invasion. Our work adds to the current understanding of parasite signaling processes required for invasion, highlighting PKA as a potential drug target to inhibit invasion for the treatment of malaria.Mary-Louise WildeTony TrigliaDanushka MarapanaJennifer K. ThompsonAlexei A. KouzmitchevHayley E. BullenPaul R. GilsonAlan F. CowmanChristopher J. TonkinAmerican Society for MicrobiologyarticleAMP-activated kinasesPlasmodium falciparumhost cell invasionmalariaMicrobiologyQR1-502ENmBio, Vol 10, Iss 5 (2019)
institution DOAJ
collection DOAJ
language EN
topic AMP-activated kinases
Plasmodium falciparum
host cell invasion
malaria
Microbiology
QR1-502
spellingShingle AMP-activated kinases
Plasmodium falciparum
host cell invasion
malaria
Microbiology
QR1-502
Mary-Louise Wilde
Tony Triglia
Danushka Marapana
Jennifer K. Thompson
Alexei A. Kouzmitchev
Hayley E. Bullen
Paul R. Gilson
Alan F. Cowman
Christopher J. Tonkin
Protein Kinase A Is Essential for Invasion of <named-content content-type="genus-species">Plasmodium falciparum</named-content> into Human Erythrocytes
description ABSTRACT Understanding the mechanisms behind host cell invasion by Plasmodium falciparum remains a major hurdle to developing antimalarial therapeutics that target the asexual cycle and the symptomatic stage of malaria. Host cell entry is enabled by a multitude of precisely timed and tightly regulated receptor-ligand interactions. Cyclic nucleotide signaling has been implicated in regulating parasite invasion, and an important downstream effector of the cAMP-signaling pathway is protein kinase A (PKA), a cAMP-dependent protein kinase. There is increasing evidence that P. falciparum PKA (PfPKA) is responsible for phosphorylation of the cytoplasmic domain of P. falciparum apical membrane antigen 1 (PfAMA1) at Ser610, a cAMP-dependent event that is crucial for successful parasite invasion. In the present study, CRISPR-Cas9 and conditional gene deletion (dimerizable cre) technologies were implemented to generate a P. falciparum parasite line in which expression of the catalytic subunit of PfPKA (PfPKAc) is under conditional control, demonstrating highly efficient dimerizable Cre recombinase (DiCre)-mediated gene excision and complete knockdown of protein expression. Parasites lacking PfPKAc show severely reduced growth after one intraerythrocytic growth cycle and are deficient in host cell invasion, as highlighted by live-imaging experiments. Furthermore, PfPKAc-deficient parasites are unable to phosphorylate PfAMA1 at Ser610. This work not only identifies an essential role for PfPKAc in the P. falciparum asexual life cycle but also confirms that PfPKAc is the kinase responsible for phosphorylating PfAMA1 Ser610. IMPORTANCE Malaria continues to present a major global health burden, particularly in low-resource countries. Plasmodium falciparum, the parasite responsible for the most severe form of malaria, causes disease through rapid and repeated rounds of invasion and replication within red blood cells. Invasion into red blood cells is essential for P. falciparum survival, and the molecular events mediating this process have gained much attention as potential therapeutic targets. With no effective vaccine available, and with the emergence of resistance to antimalarials, there is an urgent need for the development of new therapeutics. Our research has used genetic techniques to provide evidence of an essential protein kinase involved in P. falciparum invasion. Our work adds to the current understanding of parasite signaling processes required for invasion, highlighting PKA as a potential drug target to inhibit invasion for the treatment of malaria.
format article
author Mary-Louise Wilde
Tony Triglia
Danushka Marapana
Jennifer K. Thompson
Alexei A. Kouzmitchev
Hayley E. Bullen
Paul R. Gilson
Alan F. Cowman
Christopher J. Tonkin
author_facet Mary-Louise Wilde
Tony Triglia
Danushka Marapana
Jennifer K. Thompson
Alexei A. Kouzmitchev
Hayley E. Bullen
Paul R. Gilson
Alan F. Cowman
Christopher J. Tonkin
author_sort Mary-Louise Wilde
title Protein Kinase A Is Essential for Invasion of <named-content content-type="genus-species">Plasmodium falciparum</named-content> into Human Erythrocytes
title_short Protein Kinase A Is Essential for Invasion of <named-content content-type="genus-species">Plasmodium falciparum</named-content> into Human Erythrocytes
title_full Protein Kinase A Is Essential for Invasion of <named-content content-type="genus-species">Plasmodium falciparum</named-content> into Human Erythrocytes
title_fullStr Protein Kinase A Is Essential for Invasion of <named-content content-type="genus-species">Plasmodium falciparum</named-content> into Human Erythrocytes
title_full_unstemmed Protein Kinase A Is Essential for Invasion of <named-content content-type="genus-species">Plasmodium falciparum</named-content> into Human Erythrocytes
title_sort protein kinase a is essential for invasion of <named-content content-type="genus-species">plasmodium falciparum</named-content> into human erythrocytes
publisher American Society for Microbiology
publishDate 2019
url https://doaj.org/article/e57a13863f35463ca89cb2fe9e8b1b58
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