The malarial CDK Pfmrk and its effector PfMAT1 phosphorylate DNA replication proteins and co-localize in the nucleus

Jirage, Dayadevi; Chen, Yue‐Qin; Caridha, Diana; O’Neil, Michael T.; Eyase, Fredrick; Witola, William H.; Mamoun, Choukri Ben; Waters, Norman C.

doi:10.1016/j.molbiopara.2010.03.009

Cited by 26 publications

(22 citation statements)

References 59 publications

(88 reference statements)

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“…Individual pairwise interactions have been demonstrated previously (16). However, this is the first biochemical identification of the trimeric complex of PfCyc1/PfMAT1/PfMRK in an apicomplexan species, and the first isolation of this complex from live parasites.…”

Section: Observationmentioning

confidence: 53%

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The Malaria Parasite Cyclin H Homolog PfCyc1 Is Required for Efficient Cytokinesis in Blood-Stage Plasmodium falciparum

Robbins

Absalon

Streva

et al. 2017

mBio

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All well-studied eukaryotic cell cycles are driven by cyclins, which activate cyclin-dependent kinases (CDKs), and these protein kinase complexes are viable drug targets. The regulatory control of the Plasmodium falciparum cell division cycle remains poorly understood, and the roles of the various CDKs and cyclins remain unclear. The P. falciparum genome contains multiple CDKs, but surprisingly, it does not contain any sequence-identifiable G1-, S-, or M-phase cyclins. We demonstrate that P. falciparum Cyc1 (PfCyc1) complements a G1 cyclin-depleted Saccharomyces cerevisiae strain and confirm that other identified malaria parasite cyclins do not complement this strain. PfCyc1, which has the highest sequence similarity to the conserved cyclin H, cannot complement a temperature-sensitive yeast cyclin H mutant. Coimmunoprecipitation of PfCyc1 from P. falciparum parasites identifies PfMAT1 and PfMRK as specific interaction partners and does not identify PfPK5 or other CDKs. We then generate an endogenous conditional allele of PfCyc1 in blood-stage P. falciparum using a destabilization domain (DD) approach and find that PfCyc1 is essential for blood-stage proliferation. PfCyc1 knockdown does not impede nuclear division, but it prevents proper cytokinesis. Thus, we demonstrate that PfCyc1 has a functional divergence from bioinformatic predictions, suggesting that the malaria parasite cell division cycle has evolved to use evolutionarily conserved proteins in functionally novel ways.

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Section: Observationmentioning

confidence: 53%

“…PfMRK interacts with PfMAT1 in vitro , and PfCyc1 stimulates PfMRK kinase activity (16). Surprisingly, PfMRK has been found to interact with DNA replication proteins by a bacterial two-hybrid assay (16). This suggests the possibility of nonconserved functions for this kinase complex.…”

Section: Observationmentioning

confidence: 99%

The Malaria Parasite Cyclin H Homolog PfCyc1 Is Required for Efficient Cytokinesis in Blood-Stage Plasmodium falciparum

Robbins

Absalon

Streva

et al. 2017

mBio

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“…Also, Pf MRK is inhibited by bromohydrosulfonylacetamides which possess moderate antimalarial activity against drug resistant parasites, but not broad spectrum CDK inhibitors [35]. Pf MRK and its effector Pf MAT1 (where the C -terminal domain of Pf MAT1 is the Pf MRK activator domain) have been demonstrated to be co-localized to the parasite nucleus and that Pf MRK phosphorylates two plasmodial DNA replication proteins suggesting that Pf MRK in the nucleus is involved with the regulation of the DNA replication machinery [36]. CDK enzyme, cgd6_1420 is half the size of its orthologue, Pf CRK-3 (PFD0740w) with 1339 residues, which has been implicated as crucial in intraerythrocytic development of P. falciparum via regulation of gene expression [37].…”

Section: Resultsmentioning

confidence: 99%

The Cryptosporidium parvum Kinome

et al. 2011

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BackgroundHundreds of millions of people are infected with cryptosporidiosis annually, with immunocompromised individuals suffering debilitating symptoms and children in socioeconomically challenged regions at risk of repeated infections. There is currently no effective drug available. In order to facilitate the pursuit of anti-cryptosporidiosis targets and compounds, our study spans the classification of the Cryptosporidium parvum kinome and the structural and biochemical characterization of representatives from the CDPK family and a MAP kinase.ResultsThe C. parvum kinome comprises over 70 members, some of which may be promising drug targets. These C. parvum protein kinases include members in the AGC, Atypical, CaMK, CK1, CMGC, and TKL groups; however, almost 35% could only be classified as OPK (other protein kinases). In addition, about 25% of the kinases identified did not have any known orthologues outside of Cryptosporidium spp. Comparison of specific kinases with their Plasmodium falciparum and Toxoplasma gondii orthologues revealed some distinct characteristics within the C. parvum kinome, including potential targets and opportunities for drug design. Structural and biochemical analysis of 4 representatives of the CaMK group and a MAP kinase confirms features that may be exploited in inhibitor design. Indeed, screening CpCDPK1 against a library of kinase inhibitors yielded a set of the pyrazolopyrimidine derivatives (PP1-derivatives) with IC50 values of < 10 nM. The binding of a PP1-derivative is further described by an inhibitor-bound crystal structure of CpCDPK1. In addition, structural analysis of CpCDPK4 identified an unprecedented Zn-finger within the CDPK kinase domain that may have implications for its regulation.ConclusionsIdentification and comparison of the C. parvum protein kinases against other parasitic kinases shows how orthologue- and family-based research can be used to facilitate characterization of promising drug targets and the search for new drugs.

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“…We identified kinases or their accessory factors of seven PK systems, six of which have been implicated in nuclear roles in Plasmodium or other eukaryotes: PfMAT1 (PFE0610c) [88]; the α- and β-subunits of casein kinase 2 (PF11_0096, PF11_0048) [89-91]; casein kinase 1 (PF11_0377) [92,93]; the catalytic and regulatory subunits of cAMP-dependent protein kinase (PFI1685w, PFL1110c) [94]; NIMA-related protein kinase PfNEK-1 (PFL1370w) [95,96]; and mitogen-activated protein kinase 2 PfMAP2 (PF11_0147) [97,98]. PfMAP2 is a substrate of PfNEK-1 [96] and both kinases are essential for completion of the IDC in P. falciparum [95,99], whereas in P. berghei PbMAP2 is implicated specifically in the development of male gametes [97].…”

Section: Discussionmentioning

confidence: 99%

Organellar proteomics reveals hundreds of novel nuclear proteins in the malaria parasite Plasmodium falciparum

et al. 2012

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BackgroundThe post-genomic era of malaria research provided unprecedented insights into the biology of Plasmodium parasites. Due to the large evolutionary distance to model eukaryotes, however, we lack a profound understanding of many processes in Plasmodium biology. One example is the cell nucleus, which controls the parasite genome in a development- and cell cycle-specific manner through mostly unknown mechanisms. To study this important organelle in detail, we conducted an integrative analysis of the P. falciparum nuclear proteome.ResultsWe combined high accuracy mass spectrometry and bioinformatic approaches to present for the first time an experimentally determined core nuclear proteome for P. falciparum. Besides a large number of factors implicated in known nuclear processes, one-third of all detected proteins carry no functional annotation, including many phylum- or genus-specific factors. Importantly, extensive experimental validation using 30 transgenic cell lines confirmed the high specificity of this inventory, and revealed distinct nuclear localization patterns of hitherto uncharacterized proteins. Further, our detailed analysis identified novel protein domains potentially implicated in gene transcription pathways, and sheds important new light on nuclear compartments and processes including regulatory complexes, the nucleolus, nuclear pores, and nuclear import pathways.ConclusionOur study provides comprehensive new insight into the biology of the Plasmodium nucleus and will serve as an important platform for dissecting general and parasite-specific nuclear processes in malaria parasites. Moreover, as the first nuclear proteome characterized in any protist organism, it will provide an important resource for studying evolutionary aspects of nuclear biology.

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The malarial CDK Pfmrk and its effector PfMAT1 phosphorylate DNA replication proteins and co-localize in the nucleus

Cited by 26 publications

References 59 publications

The Malaria Parasite Cyclin H Homolog PfCyc1 Is Required for Efficient Cytokinesis in Blood-Stage Plasmodium falciparum

The Malaria Parasite Cyclin H Homolog PfCyc1 Is Required for Efficient Cytokinesis in Blood-Stage Plasmodium falciparum

The Cryptosporidium parvum Kinome

Organellar proteomics reveals hundreds of novel nuclear proteins in the malaria parasite Plasmodium falciparum

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