scholarly journals MRE11 is crucial for malaria transmission and its absence affects expression of interconnected networks of key genes essential for life

2020 ◽  
Author(s):  
David S. Guttery ◽  
Abhinay Ramaprasad ◽  
David J. P. Ferguson ◽  
Mohammad Zeeshan ◽  
Rajan Pandey ◽  
...  

AbstractThe 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 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-sulphur 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.

Cells ◽  
2020 ◽  
Vol 9 (12) ◽  
pp. 2590
Author(s):  
David S. Guttery ◽  
Abhinay Ramaprasad ◽  
David J. P. Ferguson ◽  
Mohammad Zeeshan ◽  
Rajan Pandey ◽  
...  

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.


2016 ◽  
Vol 113 (26) ◽  
pp. 7183-7188 ◽  
Author(s):  
Jorge M. Santos ◽  
Neuza Duarte ◽  
Jessica Kehrer ◽  
Jai Ramesar ◽  
M. Cristina Avramut ◽  
...  

Transmission of the malaria parasite from the mammalian host to the mosquito vector requires the formation of adequately adapted parasite forms and stage-specific organelles. Here we show that formation of the crystalloid—a unique and short-lived organelle of the Plasmodium ookinete and oocyst stage required for sporogony—is dependent on the precisely timed expression of the S-acyl-transferase DHHC10. DHHC10, translationally repressed in female Plasmodium berghei gametocytes, is activated translationally during ookinete formation, where the protein is essential for the formation of the crystalloid, the correct targeting of crystalloid-resident protein LAP2, and malaria parasite transmission.


2021 ◽  
Author(s):  
Rabiatou A. Diarra ◽  
Mohamed M. Traore ◽  
Amy M Junnila ◽  
Sekou F. Traore ◽  
Seydou Doumbia ◽  
...  

Abstract Background Attractive Toxic Sugar Baits (ATSBs) successfully reduced Anopheles mosquito vector populations and malaria parasite transmission in Mali, but application methods need to be improved for wide-scale use, and effects on non-target organisms (NTOs) must be assessed. The goals of this study were to determine on a village level the effect of different outdoor configurations of ATSB bait stations to 1) achieve > 25% Anopheles mosquito vector daily feeding rate for both males and females and 2) minimize the effect on non-target organisms. Methods Dye was added to Attractive Sugar Bait Stations (ASB – without toxin) to mark mosquitoes feeding on the sugar baits, and CDC UV light traps were used to monitor mosquitoes for the presence of the dye. Yellow plates, pitfall traps, Malaise traps, UV light traps, UV tray traps, and sweep nets were used to trap and sample non-target organisms (NTOs) for dye, indicating feeding on the ASB. ASB stations were hung on outer walls of village homes to determine the impact of different densities of ASBs (1,2, or 3 per home) as well as the impact of ASB height (1 m or 1.8 m above the ground on sugar feeding by anophelines. These experiments were carried out separately, on consecutive nights for mosquito and NTO monitoring. Eight villages in the Koulikoro province were chosen as the experimental locations. Results The use of one ASB station per house marked 23.11% of female and 7.11% of male An. gambiae s.l. While two and three ASB stations per house gave feeding rates above the 25% goal, there was no statistical difference in the percentage of marked mosquitoes (p=0.3141 females; p=0.9336 males). There was no difference in sugar feeding on ASB stations when hung at 1.0 and 1.8 m and (p=0.5170 females; p=0.9934 males); however, ASBs at 1.8 m had less accidental damage from village residents and animals, and subsequent invasion of non-targets through rips or holes produced. ASB stations at 1.8 m above ground were fed on by three of seven monitored insect orders. Feeding rates were less than 0.015% of total trap catches and as low as 0.0001%. The monitored orders were: Hymenoptera [ants (Formicidae), bees (Apidae), and wasps (Vespidae)], Lepidoptera (Rhopalocera, Bombyces, Geometroidea, Noctuoidea, Sphingidae, Pyraloidea), Coleoptera (Carabidae, Tenebrionidae, Scarabaeidae, Cerambycidae, and Chrysomelidae), Diptera (Brachycera, Chironomidae), Hemiptera (Cicadomorpha and Heteroptera), Neuroptera (Myrmeleontiformia) and Orthoptera (Caelifera and Ensifera). Using one or two stations limited evidence of NTO feeding to ants (Hymenoptera), Brachycera, Heteroptera, Noctuiodea, Rhopalocera, wasps (Vespidae) and wild bees (Apidae) (both Hymenoptera) and had a significantly reduced percentage of stained individuals compared to three stations which had the highest feeding rates amongst NTOs. The percentages of stained individuals were as follows: 6.84 ± 2.03% Brachycera were stained followed by wasps (Hymenoptera: Vespidae) 5.32 ± 2.27%, and Rhopalocera 2.22 ± 1.79%. Hanging the optimal number of stations per house for catching mosquitoes (two) 1.8 m above ground, limited the groups of non-targets to Brachycera, Chironomidae, Noctuoidea, Rhopalocera, parasitic wasps and wasps (both Hymenoptera: Vespidae). The three most commonly stained non-target insect groups at this height were wasps (Vespidae) (1.65 ± 0.75%), Chironomidae (0.99 ± 0.37), and Brachycera (1.55 ± 0.69%). Feeding at this height only occurred when stations were damaged.Conclusions The goal of marking one quarter of the total Anopheles mosquito vector population per day was obtained using 2 bait stations at 1.8 m height above the ground on the outer walls of houses. This configuration of ATSB stations also had minimal effects on non-target insects: only 0.0001% to 0.013% of specimens (in three orders) were marked. Stations hung 1.8 m above the ground had less accidental damage from passing people and livestock. The minimal marking of non-target insects may be attributed to visual orientation of non-mosquito insects while mosquitoes, are mostly guided by olfactory cues. Furthermore, the bait stations have a membrane cover, which if intact, is impenetrable to most sugar feeding non-target insects but is pierced by the stylets of the mosquito proboscis. Thus, most non-target insects are not exposed to the toxin even if they approach the bait stations.


mBio ◽  
2015 ◽  
Vol 6 (2) ◽  
Author(s):  
Viswanathan Lakshmanan ◽  
Matthew E. Fishbaugher ◽  
Bob Morrison ◽  
Michael Baldwin ◽  
Michael Macarulay ◽  
...  

ABSTRACT Transmission of malaria occurs during Anopheles mosquito vector blood meals, when Plasmodium sporozoites that have invaded the mosquito salivary glands are delivered to the mammalian host. Sporozoites display a unique form of motility that is essential for their movement across cellular host barriers and invasion of hepatocytes. While the molecular machinery powering motility and invasion is increasingly well defined, the signaling events that control these essential parasite activities have not been clearly delineated. Here, we identify a phosphodiesterase (PDEγ) in Plasmodium, a regulator of signaling through cyclic nucleotide second messengers. Reverse transcriptase PCR (RT-PCR) analysis and epitope tagging of endogenous PDEγ detected its expression in blood stages and sporozoites of Plasmodium yoelii. Deletion of PDEγ (pdeγ−) rendered sporozoites nonmotile, and they failed to invade the mosquito salivary glands. Consequently, PDEγ deletion completely blocked parasite transmission by mosquito bite. Strikingly, pdeγ− sporozoites showed dramatically elevated levels of cyclic GMP (cGMP), indicating that a perturbation in cyclic nucleotide balance is involved in the observed phenotypic defects. Transcriptome sequencing (RNA-Seq) analysis of pdeγ− sporozoites revealed reduced transcript abundance of genes that encode key components of the motility and invasion apparatus. Our data reveal a crucial role for PDEγ in maintaining the cyclic nucleotide balance in the malaria parasite sporozoite stage, which in turn is essential for parasite transmission from mosquito to mammal. IMPORTANCE Malaria is a formidable threat to human health worldwide, and there is an urgent need to identify novel drug targets for this parasitic disease. The parasite is transmitted by mosquito bite, inoculating the host with infectious sporozoite stages. We show that cellular signaling by cyclic nucleotides is critical for transmission of the parasite from the mosquito vector to the mammalian host. Parasite phosphodiesterase γ is essential for maintaining cyclic nucleotide balance, and its deletion blocks transmission of sporozoites. A deeper understanding of the signaling mechanisms involved in transmission might inform the discovery of novel drugs that interrupt this essential step in the parasite life cycle.


2018 ◽  
Vol 9 (1) ◽  
Author(s):  
Jung-Hee Lee ◽  
Seon-Joo Park ◽  
Gurusamy Hariharasudhan ◽  
Min-Ji Kim ◽  
Sung Mi Jung ◽  
...  

2016 ◽  
Vol 113 (26) ◽  
pp. E3676-E3685 ◽  
Author(s):  
Nicholas A. Willis ◽  
Chunshui Zhou ◽  
Andrew E. H. Elia ◽  
Johanne M. Murray ◽  
Antony M. Carr ◽  
...  

The cellular response to DNA damage during S-phase regulates a complicated network of processes, including cell-cycle progression, gene expression, DNA replication kinetics, and DNA repair. In fission yeast, this S-phase DNA damage response (DDR) is coordinated by two protein kinases: Rad3, the ortholog of mammalian ATR, and Cds1, the ortholog of mammalian Chk2. Although several critical downstream targets of Rad3 and Cds1 have been identified, most of their presumed targets are unknown, including the targets responsible for regulating replication kinetics and coordinating replication and repair. To characterize targets of the S-phase DDR, we identified proteins phosphorylated in response to methyl methanesulfonate (MMS)-induced S-phase DNA damage in wild-type, rad3∆, and cds1∆ cells by proteome-wide mass spectrometry. We found a broad range of S-phase–specific DDR targets involved in gene expression, stress response, regulation of mitosis and cytokinesis, and DNA replication and repair. These targets are highly enriched for proteins required for viability in response to MMS, indicating their biological significance. Furthermore, the regulation of these proteins is similar in fission and budding yeast, across 300 My of evolution, demonstrating a deep conservation of S-phase DDR targets and suggesting that these targets may be critical for maintaining genome stability in response to S-phase DNA damage across eukaryotes.


2003 ◽  
Vol 23 (12) ◽  
pp. 4247-4256 ◽  
Author(s):  
Zhi-Xiang Xu ◽  
Anna Timanova-Atanasova ◽  
Rui-Xun Zhao ◽  
Kun-Sang Chang

ABSTRACT The PML tumor suppressor gene is consistently disrupted by t(15;17) in patients with acute promyelocytic leukemia. Promyelocytic leukemia protein (PML) is a multifunctional protein that plays essential roles in cell growth regulation, apoptosis, transcriptional regulation, and genome stability. Our study here shows that PML colocalizes and associates in vivo with the DNA damage response protein TopBP1 in response to ionizing radiation (IR). Both PML and TopBP1 colocalized with the IR-induced bromodeoxyuridine single-stranded DNA foci. PML and TopBP1 also colocalized with Rad50, Brca1, ATM, Rad9, and BLM. IR and interferon (IFN) coinduce the expression levels of both TopBP1 and PML. In PML-deficient NB4 cells, TopBP1 was unable to form IR-induced foci. All-trans-retinoic acid induced reorganization of the PML nuclear body (NB) and reappearance of the IR-induced TopBP1 foci. Inhibition of PML expression by siRNA is associated with a significant decreased in TopBP1 expression. Furthermore, PML-deficient cells express a low level of TopBP1, and its expression cannot be induced by IR or IFN. Adenovirus-mediated overexpression of PML in PML−/− mouse embryo fibroblasts substantially increased TopBP1 expression, which colocalized with the PML NBs. These studies demonstrated a mechanism of PML-dependent expression of TopBP1. PML overexpression induced TopBP1 protein but not the mRNA expression. Pulse-chase labeling analysis demonstrated that PML overexpression stabilized the TopBP1 protein, suggesting that PML plays a role in regulating the stability of TopBP1 in response to IR. Together, our findings demonstrate that PML regulates TopBP1 functions by association and stabilization of the protein in response to IR-induced DNA damage.


2020 ◽  
Author(s):  
Rabiatou A. Diarra A. Diarra ◽  
Mohamed M. Traore ◽  
Amy M Junnila ◽  
Sekou F. Traore ◽  
Seydou Doumbia ◽  
...  

Abstract Background Attractive Toxic Sugar Baits (ATSBs) successfully reduced Anopheles mosquito vector populations and malaria parasite transmission in Mali but application methods need to be improved for wide-scale use, and minimization of effects on non-target organisms (NTOs) must be assessed. The goals of this study were to determine on a village level the effect of different outdoor configurations of ATSB bait stations to 1) achieve > 25% Anopheles mosquito vector daily feeding rate for both males and females and 2) minimize the effect on non-target organisms. Methods Dye was added to Attractive Sugar Bait Stations (ASB – without toxin) to mark mosquitoes feeding on the sugar baits, and CDC UV light traps were used to monitor mosquitoes for the presence of the dye. Yellow plates, pitfall traps, Malaise traps, UV light traps, UV tray traps, and sweep nets were used to trap and sample non-target organisms (NTOs) for dye, indicating feeding on the ASB. ASB stations were hung on outer walls of village homes to determine the impact of different densities of ASBs (1,2, or 3 per home) as well as the impact of ASB height (1 m or 1.8 m above the ground on sugar feeding by anophelines. These experiments were carried out separately, on consecutive nights for mosquito and NTO monitoring. Eight villages in the Koulikoro province were chosen as the experimental locations Results The use of one ASB station per house marked 23.11% of female and 14.64% of male An. gambiae s.l. While two and three ASB stations per house gave feeding rates above the 25% goal, there was no statistical difference in the percentage of marked mosquitoes (p = 0.3141 females; p = 0.9336 males). There was no difference in sugar feeding on ASB stations when hung at 1.0 and 1.8 m and (p = 0.5170 females; p = 0.9934 males); however, ASBs at 1.8 m had less accidental damage from village residents and animals, and subsequent invasion of non-targets through rips or holes produced. ASB stations at 1.8 m above ground were fed on by three of seven monitored insect orders. Feeding rates were less than 0.015% of total trap catches and as low as 0.0001%. The monitored orders were: Hymenoptera [ants (Formicidae), bees (Apidae), and wasps (Vespidae)], Lepidoptera (Rhopalocera, Bombyces, Geometroidea, Noctuoidea, Sphingidae, Pyraloidea), Coleoptera (Carabidae, Tenebrionidae, Scarabaeidae, Cerambycidae, and Chrysomelidae), Diptera (Brachycera, Chironomidae), Hemiptera (Cicadomorpha and Heteroptera), Neuroptera (Myrmeleontiformia) and Orthoptera (Caelifera and Ensifera). Using one or two stations limited evidence of NTO feeding to ants (Hymenoptera), Brachycera, Heteroptera, Noctuiodea, Rhopalocera, wasps (Vespidae) and wild bees (Apidae) (both Hymenoptera) and had a significantly reduced percentage of stained individuals compared to three stations which produced the highest number of NTOs. The percentages of stained individuals were as follows: Brachycera accounted for 6.84 ± 2.03% of stained insects, wasps (Hymenoptera: Vespidae) for 5.32 ± 2.27%, and Rhopalocera for 2.22 ± 1.79%. Hanging the optimal number of stations per house for catching mosquitoes (two) 1.8 m above ground limited the groups of non-targets to Brachycera, Chironomidae, Noctuoidea, Rhopalocera, parasitic wasps and wasps (both Hymenoptera: Vespidae). The three most commonly stained non-target insect groups at this height were wasps (Vespidae) (1.65 ± 0.75%), Chironomidae (0.99 ± 0.37), and Brachycera (1.55 ± 0.69%). Feeding at this height only occurred when stations were damaged. Conclusions The goal of marking one quarter of the total Anopheles mosquito vector population per day was obtained using 2 bait stations at 1.8 m height above the ground on the outer walls of houses. This configuration of ATSB stations also had minimal effects on non-target insects: only 0.0001–0.013% of specimens (in three orders) were marked. Stations hung 1.8 m above the ground had less accidental damage from passing people and livestock. The minimal marking of non-target insects may be attributed to visual orientation of non-mosquito insects while mosquitoes, are mostly guided by olfactory cues. Furthermore, the bait stations have a membrane cover, which if intact, is impenetrable to most sugar feeding non-target insects but is pierced by the stylets of the mosquito proboscis. Thus, most non-target insects are not exposed to the toxin even if they approach the bait stations.


2014 ◽  
Vol 25 (10) ◽  
pp. 1641-1652 ◽  
Author(s):  
Mattia Poletto ◽  
Lisa Lirussi ◽  
David M. Wilson ◽  
Gianluca Tell

Nucleophosmin (NPM1) is a multifunctional protein that controls cell growth and genome stability via a mechanism that involves nucleolar–cytoplasmic shuttling. It is clear that NPM1 also contributes to the DNA damage response, yet its exact function is poorly understood. We recently linked NPM1 expression to the functional activation of the major abasic endonuclease in mammalian base excision repair (BER), apurinic/apyrimidinic endonuclease 1 (APE1). Here we unveil a novel role for NPM1 as a modulator of the whole BER pathway by 1) controlling BER protein levels, 2) regulating total BER capacity, and 3) modulating the nucleolar localization of several BER enzymes. We find that cell treatment with the genotoxin cisplatin leads to concurrent relocalization of NPM1 and BER components from nucleoli to the nucleoplasm, and cellular experiments targeting APE1 suggest a role for the redistribution of nucleolar BER factors in determining cisplatin toxicity. Finally, based on the use of APE1 as a representative protein of the BER pathway, our data suggest a function for BER proteins in the regulation of ribogenesis.


Cells ◽  
2019 ◽  
Vol 8 (10) ◽  
pp. 1157
Author(s):  
Hisako Fujihara ◽  
Tadashige Nozaki ◽  
Masahiro Tsutsumi ◽  
Mayu Isumi ◽  
Shinji Shimoda ◽  
...  

Poly(ADP-ribose) polymerase (Parp)-1 catalyzes polyADP-ribosylation using NAD+ and is involved in the DNA damage response, genome stability, and transcription. In this study, we demonstrated that aged Parp-1−/− mouse incisors showed more frequent dental dysplasia in both ICR/129Sv mixed background and C57BL/6 strain compared to aged Parp-1+/+ incisors, suggesting that Parp-1 deficiency could be involved in development of dental dysplasia at an advanced age. Computed tomography images confirmed that dental dysplasia was observed at significantly higher incidences in Parp-1−/− mice. The relative calcification levels of Parp-1−/− incisors were higher in both enamel and dentin (p < 0.05). Immunohistochemical analysis revealed (1) Parp-1 positivity in ameloblasts and odontoblasts in Parp-1+/+ incisor, (2) weaker dentin sialoprotein positivity in dentin of Parp-1−/− incisor, and (3) bone sialoprotein positivity in dentin of Parp-1−/− incisor, suggesting ectopic osteogenic formation in dentin of Parp-1−/− incisor. These results indicate that Parp-1 deficiency promotes odontogenic failure in incisors at an advanced age. Parp-1 deficiency did not affect dentinogenesis during the development of mice, suggesting that Parp-1 is not essential in dentinogenesis during development but is possibly involved in the regulation of continuous dentinogenesis in the incisors at an advanced age.


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