scholarly journals Plasmodium falciparum Calcium Dependent Protein Kinase 4 is critical for male gametogenesis and transmission to the mosquito vector

2021 ◽  
Author(s):  
SUDHIR KUMAR ◽  
Meseret T. Haile ◽  
Michael R. Hoopmann ◽  
Linh T. Tran ◽  
Samantha A. Michaels ◽  
...  
Keyword(s):  
Plasmodium Falciparum ◽  
Life Cycle ◽  
Mrna Translation ◽  
Complex Life Cycle ◽  
Mosquito Vector ◽  
Parasite Transmission ◽  
Calcium Dependent ◽  
Male Gametogenesis ◽  
Parasite Plasmodium ◽  
Dependent Protein

Gametocytes of the malaria parasite Plasmodium are taken up by the mosquito vector with an infectious blood meal, representing a critical stage for parasite transmission. Calcium dependent protein kinases play key roles in calcium mediated signaling across the complex life cycle of the parasite. We sought to understand their role in human parasite transmission from the host to the mosquito vector and thus investigated the role of the human infective parasite Plasmodium falciparum CDPK4 in the parasite life cycle. P. falciparum cdpk4 parasites created by targeted gene deletion showed no effect in blood stage development or gametocyte development. However, cdpk4 parasites showed a severe defect in male gametogenesis and the emergence of flagellated male gametes. To understand the molecular underpinnings of this defect, we performed mass spectrometry based phosphoproteomic analyses of wild type and Plasmodium falciparum cdpk4 late gametocyte stages, to identify key CDPK4 mediated phosphorylation events that may be important for the regulation of male gametogenesis. We further employed in vitro assays to identify these putative substrates of Plasmodium falciparum CDPK4. This indicated that CDPK4 regulates male gametogenesis by directly or indirectly controlling key essential events such as DNA replication, mRNA translation and cell motility. Taken together, our work demonstrates that PfCDPK4 is a central kinase that regulates exflagellation, and thereby is critical for parasite transmission to the mosquito vector.

scholarly journals Plasmodium falciparum Cyclic GMP-Dependent Protein Kinase Interacts with a Subunit of the Parasite Proteasome

2018 ◽  
Vol 87 (1) ◽  
Author(s):  
K. Govindasamy ◽  
R. Khan ◽  
M. Snyder ◽  
H. J. Lou ◽  
P. Du ◽  
...  
Keyword(s):  
Plasmodium Falciparum ◽  
Life Cycle ◽  
Protein Kinase ◽  
Cyclic Gmp ◽  
Phosphorylation Site ◽  
Complex Life Cycle ◽  
Mosquito Vector ◽  
Dependent Protein Kinase ◽  
Content Type ◽  
Dependent Protein

ABSTRACT Malaria is caused by the protozoan parasite Plasmodium, which undergoes a complex life cycle in a human host and a mosquito vector. The parasite’s cyclic GMP (cGMP)-dependent protein kinase (PKG) is essential at multiple steps of the life cycle. Phosphoproteomic studies in Plasmodium falciparum erythrocytic stages and Plasmodium berghei ookinetes have identified proteolysis as a major biological pathway dependent on PKG activity. To further understand PKG’s mechanism of action, we screened a yeast two-hybrid library for P. falciparum proteins that interact with P. falciparum PKG (PfPKG) and tested peptide libraries to identify its phosphorylation site preferences. Our data suggest that PfPKG has a distinct phosphorylation site and that PfPKG directly phosphorylates parasite RPT1, one of six AAA+ ATPases present in the 19S regulatory particle of the proteasome. PfPKG and RPT1 interact in vitro, and the interacting fragment of RPT1 carries a PfPKG consensus phosphorylation site; a peptide carrying this consensus site competes with the RPT1 fragment for binding to PfPKG and is efficiently phosphorylated by PfPKG. These data suggest that PfPKG’s phosphorylation of RPT1 could contribute to its regulation of parasite proteolysis. We demonstrate that proteolysis plays an important role in a biological process known to require Plasmodium PKG: invasion by sporozoites of hepatocytes. A small-molecule inhibitor of proteasomal activity blocks sporozoite invasion in an additive manner when combined with a Plasmodium PKG-specific inhibitor. Mining the previously described parasite PKG-dependent phosphoproteomes using the consensus phosphorylation motif identified additional proteins that are likely to be direct substrates of the enzyme.

scholarly journals A Novel Tool for the Generation of Conditional Knockouts To Study Gene Function across the Plasmodium falciparum Life Cycle

mBio ◽  
2019 ◽  
Vol 10 (5) ◽  
Author(s):  
Marta Tibúrcio ◽  
Annie S. P. Yang ◽  
Kazuhide Yahata ◽  
Pablo Suárez-Cortés ◽  
Hugo Belda ◽  
...  
Keyword(s):  
Plasmodium Falciparum ◽  
Life Cycle ◽  
Genetic Modification ◽  
Gene Deletion ◽  
Genetically Modify ◽  
Complex Life Cycle ◽  
Mosquito Vector ◽  
Parasite Line ◽  
Content Type ◽  
First Time

ABSTRACT Plasmodium falciparum has a complex life cycle that involves interaction with multiple tissues inside the human and mosquito hosts. Identification of essential genes at all different stages of the P. falciparum life cycle is urgently required for clinical development of tools for malaria control and eradication. However, the study of P. falciparum is limited by the inability to genetically modify the parasite throughout its life cycle with the currently available genetic tools. Here, we describe the detailed characterization of a new marker-free P. falciparum parasite line that expresses rapamycin-inducible Cre recombinase across the full life cycle. Using this parasite line, we were able to conditionally delete the essential invasion ligand AMA1 in three different developmental stages for the first time. We further confirm efficient gene deletion by targeting the nonessential kinase FIKK7.1. IMPORTANCE One of the major limitations in studying P. falciparum is that so far only asexual stages are amenable to rapid conditional genetic modification. The most promising drug targets and vaccine candidates, however, have been refractory to genetic modification because they are essential during the blood stage or for transmission in the mosquito vector. This leaves a major gap in our understanding of parasite proteins in most life cycle stages and hinders genetic validation of drug and vaccine targets. Here, we describe a method that supports conditional gene deletion across the P. falciparum life cycle for the first time. We demonstrate its potential by deleting essential and nonessential genes at different parasite stages, which opens up completely new avenues for the study of malaria and drug development. It may also allow the realization of novel vaccination strategies using attenuated parasites.

scholarly journals Plasmodium falciparum Calcium-Dependent Protein Kinase 4 is Critical for Male Gametogenesis and Transmission to the Mosquito Vector

mBio ◽  
2021 ◽  
Author(s):  
Sudhir Kumar ◽  
Meseret T. Haile ◽  
Michael R. Hoopmann ◽  
Linh T. Tran ◽  
Samantha A. Michaels ◽  
...  
Keyword(s):  
Male Gamete ◽  
Mosquito Vector ◽  
Parasite Life Cycle ◽  
Dependent Protein Kinase ◽  
Calcium Dependent ◽  
Male Gametogenesis ◽  
Phosphorylation Of Proteins ◽  
Male Gametes ◽  
Dependent Protein ◽  
Calcium Dependent Protein Kinase

Transmission of the malaria parasite to the mosquito vector is critical for the completion of the sexual stage of the parasite life cycle and is dependent on the release of male gametes from the gametocyte body inside the mosquito midgut. In the present study, we demonstrate that PfCDPK4 is critical for male gametogenesis and is involved in phosphorylation of proteins essential for male gamete emergence.

scholarly journals Mitosis in the Human Malaria Parasite Plasmodium falciparum

2011 ◽  
Vol 10 (4) ◽  
pp. 474-482 ◽  
Author(s):  
Noel Gerald ◽  
Babita Mahajan ◽  
Sanjai Kumar
Keyword(s):  
Plasmodium Falciparum ◽  
Life Cycle ◽  
Complex Life Cycle ◽  
Anopheline Mosquito ◽  
Essential Requirement ◽  
Human Malaria Parasite ◽  
Content Type ◽  
Parasite Plasmodium ◽  
Parasite Plasmodium Falciparum ◽  
Key Events

ABSTRACT Malaria is caused by intraerythrocytic protozoan parasites belonging to Plasmodium spp. (phylum Apicomplexa ) that produce significant morbidity and mortality, mostly in developing countries. Plasmodium parasites have a complex life cycle that includes multiple stages in anopheline mosquito vectors and vertebrate hosts. During the life cycle, the parasites undergo several cycles of extreme population growth within a brief span, and this is critical for their continued transmission and a contributing factor for their pathogenesis in the host. As with other eukaryotes, successful mitosis is an essential requirement for Plasmodium reproduction; however, some aspects of Plasmodium mitosis are quite distinct and not fully understood. In this review, we will discuss the current understanding of the architecture and key events of mitosis in Plasmodium falciparum and related parasites and compare them with the traditional mitotic events described for other eukaryotes.

scholarly journals The ApiAP2 factor PfAP2-HC is an integral component of heterochromatin in the malaria parasite Plasmodium falciparum

2020 ◽  
Author(s):  
Eilidh Carrington ◽  
Roel H. M. Cooijmans ◽  
Dominique Keller ◽  
Christa G. Toenhake ◽  
Richárd Bártfai ◽  
...  
Keyword(s):  
Gene Expression ◽  
Plasmodium Falciparum ◽  
Life Cycle ◽  
Dna Binding ◽  
Regulation Of Gene Expression ◽  
Complex Life Cycle ◽  
Parasite Development ◽  
Specific Gene ◽  
Mosquito Vector ◽  
Malaria Parasites

AbstractMalaria parasites undergo a highly complex life cycle in the human host and the mosquito vector. The ApiAP2 family of sequence-specific DNA-binding proteins plays a dominant role in parasite development and life cycle progression. Of the ApiAP2 factors studied to date, most act as transcription factors regulating stage-specific gene expression. Here, we characterised a new ApiAP2 factor in Plasmodium falciparum (PF3D7_1456000) that we termed PfAP2-HC. Via detailed investigation of several single or double genetically engineered parasite lines, we demonstrate that PfAP2-HC specifically binds to heterochromatin throughout the genome. Intriguingly, PfAP2-HC does not bind DNA in vivo and recruitment of PfAP2-HC to heterochromatin is independent of its DNA-binding domain but strictly dependent on heterochromatin protein 1. Furthermore, our results suggest that PfAP2-HC functions neither in the regulation of gene expression nor in heterochromatin formation or maintenance. In summary, our findings reveal that PfAP2-HC constitutes a core component of heterochromatin in malaria parasites. They furthermore identify unexpected properties of ApiAP2 factors and suggest substantial functional divergence among the members of this important family of regulatory proteins.

scholarly journals MRE11 Is Crucial for Malaria Parasite Transmission and Its Absence Affects Expression of Interconnected Networks of Key Genes Essential for Life

Cells ◽  
2020 ◽  
Vol 9 (12) ◽  
pp. 2590
Author(s):  
David S. Guttery ◽  
Abhinay Ramaprasad ◽  
David J. P. Ferguson ◽  
Mohammad Zeeshan ◽  
Rajan Pandey ◽  
...  
Keyword(s):  
Genome Stability ◽  
Malaria Parasite ◽  
Mosquito Vector ◽  
Parasite Transmission ◽  
Biological Processes ◽  
Cluster Assembly ◽  
Iron Sulfur Cluster ◽  
Iron Sulfur ◽  
Parasite Plasmodium ◽  
Damage Response

The meiotic recombination 11 protein (MRE11) plays a key role in DNA damage response and maintenance of genome stability. However, little is known about its function during development of the malaria parasite Plasmodium. Here, we present a functional, ultrastructural and transcriptomic analysis of Plasmodium parasites lacking MRE11 during its life cycle in both mammalian and mosquito vector hosts. Genetic disruption of Plasmodium berghei mre11 (PbMRE11) results in significant retardation of oocyst development in the mosquito midgut associated with cytoplasmic and nuclear degeneration, along with concomitant ablation of sporogony and subsequent parasite transmission. Further, absence of PbMRE11 results in significant transcriptional downregulation of genes involved in key interconnected biological processes that are fundamental to all eukaryotic life including ribonucleoprotein biogenesis, spliceosome function and iron–sulfur cluster assembly. Overall, our study provides a comprehensive functional analysis of MRE11′s role in Plasmodium development during the mosquito stages and offers a potential target for therapeutic intervention during malaria parasite transmission.

scholarly journals Probing Plasmodium falciparum sexual commitment at the single-cell level

2018 ◽  
Vol 3 ◽  
pp. 70 ◽  
Author(s):  
Nicolas M.B. Brancucci ◽  
Mariana De Niz ◽  
Timothy J. Straub ◽  
Deepali Ravel ◽  
Lauriane Sollelis ◽  
...  
Keyword(s):  
Plasmodium Falciparum ◽  
Life Cycle ◽  
Single Cell ◽  
Transcriptional Profiling ◽  
Quantitative Imaging ◽  
Complex Life Cycle ◽  
Major Transitions ◽  
Transcriptional Signature ◽  
Life Cycle Stages ◽  
Parasite Populations

Background: Malaria parasites go through major transitions during their complex life cycle, yet the underlying differentiation pathways remain obscure. Here we apply single cell transcriptomics to unravel the program inducing sexual differentiation in Plasmodium falciparum. Parasites have to make this essential life-cycle decision in preparation for human-to-mosquito transmission. Methods: By combining transcriptional profiling with quantitative imaging and genetics, we defined a transcriptional signature in sexually committed cells. Results: We found this transcriptional signature to be distinct from general changes in parasite metabolism that can be observed in response to commitment-inducing conditions. Conclusions: This proof-of-concept study provides a template to capture transcriptional diversity in parasite populations containing complex mixtures of different life-cycle stages and developmental programs, with important implications for our understanding of parasite biology and the ongoing malaria elimination campaign.

scholarly journals Revisiting gametocyte biology in malaria parasites

2019 ◽  
Vol 43 (4) ◽  
pp. 401-414 ◽  
Author(s):  
Priscilla Ngotho ◽  
Alexandra Blancke Soares ◽  
Franziska Hentzschel ◽  
Fiona Achcar ◽  
Lucia Bertuccini ◽  
...  
Keyword(s):  
Plasmodium Falciparum ◽  
Blood Circulation ◽  
Malaria Parasite ◽  
Mosquito Vector ◽  
Human Malaria Parasite ◽  
Malaria Burden ◽  
Parasite Plasmodium ◽  
Parasite Plasmodium Falciparum ◽  
Low Levels ◽  
Transmission Biology

ABSTRACT Gametocytes are the only form of the malaria parasite that is transmissible to the mosquito vector. They are present at low levels in blood circulation and significant knowledge gaps exist in their biology. Recent reductions in the global malaria burden have brought the possibility of elimination and eradication, with renewed focus on malaria transmission biology as a basis for interventions. This review discusses recent insights into gametocyte biology in the major human malaria parasite, Plasmodium falciparum and related species.

Sign in / Sign up

Export Citation Format

Share Document