scholarly journals Ca2+ signals critical for egress and gametogenesis in malaria parasites depend on a multipass membrane protein that interacts with PKG

2021 ◽  
Vol 7 (13) ◽  
pp. eabe5396
Author(s):  
Aurélia C. Balestra ◽  
Konstantinos Koussis ◽  
Natacha Klages ◽  
Steven A. Howell ◽  
Helen R. Flynn ◽  
...  
Keyword(s):  
Membrane Protein ◽  
Calcium Channels ◽  
Plasmodium Berghei ◽  
Disease Transmission ◽  
Calcium Mobilization ◽  
Dependent Protein Kinase ◽  
Malaria Parasites ◽  
Cgmp Dependent Protein Kinase ◽  
Intracellular Calcium Mobilization ◽  
Dependent Protein

Calcium signaling regulated by the cGMP-dependent protein kinase (PKG) controls key life cycle transitions in the malaria parasite. However, how calcium is mobilized from intracellular stores in the absence of canonical calcium channels in Plasmodium is unknown. Here, we identify a multipass membrane protein, ICM1, with homology to transporters and calcium channels that is tightly associated with PKG in both asexual blood stages and transmission stages. Phosphoproteomic analyses reveal multiple ICM1 phosphorylation events dependent on PKG activity. Stage-specific depletion of Plasmodium berghei ICM1 prevents gametogenesis due to a block in intracellular calcium mobilization, while conditional loss of Plasmodium falciparum ICM1 is detrimental for the parasite resulting in severely reduced calcium mobilization, defective egress, and lack of invasion. Our findings suggest that ICM1 is a key missing link in transducing PKG-dependent signals and provide previously unknown insights into atypical calcium homeostasis in malaria parasites essential for pathology and disease transmission.

scholarly journals A multipass membrane protein interacts with the cGMP-dependent protein kinase to regulate critical calcium signals in malaria parasites

2020 ◽  
Author(s):  
Aurélia C. Balestra ◽  
Konstantinos Koussis ◽  
Natacha Klages ◽  
Steven A. Howell ◽  
Helen R. Flynn ◽  
...  
Keyword(s):  
Protein Kinase ◽  
Membrane Protein ◽  
Calcium Channels ◽  
Disease Transmission ◽  
Calcium Signals ◽  
Dependent Protein Kinase ◽  
Malaria Parasites ◽  
Major Function ◽  
Cgmp Dependent Protein Kinase ◽  
Dependent Protein

AbstractIn malaria parasites, all cGMP-dependent signalling is mediated through a single cGMP-dependent protein kinase (PKG), a major function of which is to control essential calcium signals. However, how PKG transmits these signals in the absence of known second messenger-dependent calcium channels or scaffolding proteins is unknown. Here we identify a polytopic membrane protein, ICM1, with homology to transporters and calcium channels that is tightly-associated with PKG in both Plasmodium falciparum asexual blood stages and P. berghei gametocytes. Phosphoproteomic analyses in both Plasmodium species reveal multiple ICM1 phosphorylation events dependent upon PKG activity. Stage-specific depletion of P. berghei ICM1 blocks gametogenesis due to the inability of mutant parasites to mobilise intracellular calcium upon PKG activation, whilst conditional loss of P. falciparum ICM1 results in reduced calcium mobilisation, defective egress and lack of invasion. Our findings provide new insights into atypical calcium homeostasis in malaria parasites essential for pathology and disease transmission.

scholarly journals Gametogenesis in Malaria Parasites Is Mediated by the cGMP-Dependent Protein Kinase

PLoS Biology ◽  
2008 ◽  
Vol 6 (6) ◽  
pp. e139 ◽  
Author(s):  
Louisa McRobert ◽  
Cathy J Taylor ◽  
Wensheng Deng ◽  
Quinton L Fivelman ◽  
Ross M Cummings ◽  
...  
Keyword(s):  
Protein Kinase ◽  
Dependent Protein Kinase ◽  
Malaria Parasites ◽  
Cgmp Dependent Protein Kinase ◽  
Dependent Protein

scholarly journals Regulation of Cloned Cardiac L-type Calcium Channels by cGMP-dependent Protein Kinase

2000 ◽  
Vol 275 (9) ◽  
pp. 6135-6143 ◽  
Author(s):  
L. H. Jiang ◽  
D. J. Gawler ◽  
N. Hodson ◽  
C. J. Milligan ◽  
H. A. Pearson ◽  
...  
Keyword(s):  
Protein Kinase ◽  
Calcium Channels ◽  
Dependent Protein Kinase ◽  
Cgmp Dependent Protein Kinase ◽  
Dependent Protein

scholarly journals Structures of the cGMP-dependent protein kinase in malaria parasites reveal a unique structural relay mechanism for activation

2019 ◽  
Vol 116 (28) ◽  
pp. 14164-14173 ◽  
Author(s):  
Majida El Bakkouri ◽  
Imène Kouidmi ◽  
Amy K. Wernimont ◽  
Mehrnaz Amani ◽  
Ashley Hutchinson ◽  
...  
Keyword(s):  
Protein Kinase ◽  
Conformational Changes ◽  
Catalytic Domain ◽  
Kinase Domain ◽  
Complex Life Cycle ◽  
Dependent Protein Kinase ◽  
Malaria Parasites ◽  
Cgmp Dependent Protein Kinase ◽  
Dependent Protein ◽  
Developmental Switches

The cyclic guanosine-3′,5′-monophosphate (cGMP)-dependent protein kinase (PKG) was identified >25 y ago; however, efforts to obtain a structure of the entire PKG enzyme or catalytic domain from any species have failed. In malaria parasites, cooperative activation of PKG triggers crucial developmental transitions throughout the complex life cycle. We have determined the cGMP-free crystallographic structures of PKG fromPlasmodium falciparumandPlasmodium vivax, revealing how key structural components, including an N-terminal autoinhibitory segment (AIS), four predicted cyclic nucleotide-binding domains (CNBs), and a kinase domain (KD), are arranged when the enzyme is inactive. The four CNBs and the KD are in a pentagonal configuration, with the AIS docked in the substrate site of the KD in a swapped-domain dimeric arrangement. We show that although the protein is predominantly a monomer (the dimer is unlikely to be representative of the physiological form), the binding of the AIS is necessary to keepPlasmodiumPKG inactive. A major feature is a helix serving the dual role of the N-terminal helix of the KD as well as the capping helix of the neighboring CNB. A network of connecting helices between neighboring CNBs contributes to maintaining the kinase in its inactive conformation. We propose a scheme in which cooperative binding of cGMP, beginning at the CNB closest to the KD, transmits conformational changes around the pentagonal molecule in a structural relay mechanism, enabling PKG to orchestrate rapid, highly regulated developmental switches in response to dynamic modulation of cGMP levels in the parasite.

scholarly journals Role of Plasmodium berghei cGMP-dependent Protein Kinase in Late Liver Stage Development

2009 ◽  
Vol 285 (5) ◽  
pp. 3282-3288 ◽  
Author(s):  
Adebola Falae ◽  
Audrey Combe ◽  
Anburaj Amaladoss ◽  
Teresa Carvalho ◽  
Robert Menard ◽  
...  
Keyword(s):  
Protein Kinase ◽  
Plasmodium Berghei ◽  
Dependent Protein Kinase ◽  
Liver Stage ◽  
Cgmp Dependent Protein Kinase ◽  
Dependent Protein ◽  
Stage Development

scholarly journals Simultaneous multiple allelic replacement in the malaria parasite enables dissection of PKG function

2020 ◽  
Vol 3 (4) ◽  
pp. e201900626 ◽  
Author(s):  
Konstantinos Koussis ◽  
Chrislaine Withers-Martinez ◽  
David A Baker ◽  
Michael J Blackman
Keyword(s):  
Plasmodium Falciparum ◽  
Gene Function ◽  
Disease Transmission ◽  
Malaria Parasite ◽  
Gene Copy ◽  
Dependent Protein Kinase ◽  
Cgmp Dependent Protein Kinase ◽  
Allelic Replacement ◽  
Dependent Protein ◽  
Multiple Recombination

Over recent years, a plethora of new genetic tools has transformed conditional engineering of the malaria parasite genome, allowing functional dissection of essential genes in the asexual and sexual blood stages that cause pathology or are required for disease transmission, respectively. Important challenges remain, including the desirability to complement conditional mutants with a correctly regulated second gene copy to confirm that observed phenotypes are due solely to loss of gene function and to analyse structure–function relationships. To meet this challenge, here we combine the dimerisable Cre (DiCre) system with the use of multiple lox sites to simultaneously generate multiple recombination events of the same gene. We focused on the Plasmodium falciparum cGMP-dependent protein kinase (PKG), creating in parallel conditional disruption of the gene plus up to two allelic replacements. We use the approach to demonstrate that PKG has no scaffolding or adaptor role in intraerythrocytic development, acting solely at merozoite egress. We also show that a phosphorylation-deficient PKG is functionally incompetent. Our method provides valuable new tools for analysis of gene function in the malaria parasite.

cDNA Cloning and Gene Expression of Human Type Iα cGMP-Dependent Protein Kinase

Hypertension ◽  
1996 ◽  
Vol 27 (3) ◽  
pp. 552-557 ◽  
Author(s):  
Naohisa Tamura ◽  
Hiroshi Itoh ◽  
Yoshihiro Ogawa ◽  
Osamu Nakagawa ◽  
Masaki Harada ◽  
...  
Keyword(s):  
Gene Expression ◽  
Protein Kinase ◽  
Cdna Cloning ◽  
Dependent Protein Kinase ◽  
Human Type ◽  
Cgmp Dependent Protein Kinase ◽  
Dependent Protein
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