Dataset Information

Identification of nuclear Plasmodium falciparum proteins via mass spectrometry and proposed annotation

ABSTRACT: The nuclear proteome of mixed populations of Plasmodium falciparum was explored by GeLC-MS/MS and 2D-LC-MS/MS with a peptide exclusion procedure in order to increase the detection of low abundant proteins such as those involved in gene expression.

INSTRUMENT(S): HCTultra

ORGANISM(S): Plasmodium Falciparum (isolate 3d7)

SUBMITTER: Cédric Pionneau

LAB HEAD: Solenne Chardonnet

PROVIDER: PXD010427 | Pride | 2019-04-02

REPOSITORIES: pride

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Identification of Plasmodium falciparum nuclear proteins by mass spectrometry and proposed protein annotation.

Briquet Sylvie S Ourimi Asma A Pionneau Cédric C Bernardes Juliana J Carbone Alessandra A Chardonnet Solenne S Vaquero Catherine C

PloS one 20181031 10

The nuclear proteome of Plasmodium falciparum results from the continual shuttle of proteins between the cell cytoplasm-nucleus and vice versa. Using shotgun proteomics tools, we explored the nuclear proteins of mixed populations of Plasmodium falciparum extracted from infected erythrocytes. We combined GeLC-MS/MS and 2D-LC-MS/MS with a peptide ion exclusion procedure in order to increase the detection of low abundant proteins such as those involved in gene expression. We have identified 446 nuc ...[more]

PMID: 30379851

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Project description:For malaria transmission, the parasite must undergo sexual differentiation into mature gametocytes. However, the molecular basis for this critical transition in the parasites life cycle is unknown. Six previously uncharacterized genes, Pfg14.744, Pfg14.745, Pfg14.748, Pfg14.763, Pfg14.752 and Pfg6.6 that are members of a 36 gene Plasmodium falciparum-specific subtelomeric superfamily were found to be expressed in parasites that are committed to sexual development as suggested by co-expression of Pfs16 and Pfg27. Northern blots demonstrated that Pfg14.744 and Pfg14.748 were first expressed before the parasites differentiated into morphologically distinct gametocytes, transcription continued to increase until stage II gametocytes were formed and then rapidly decreased. Immunofluorescence assays indicated that both proteins were only produced in the subpopulation of ring stage parasites that are committed to gametocytogenesis and both localized to the parasitophorous vacuole (PV)b of the early ring stage parasites. As the parasites continued to develop Pfg14.748 remained within the parasitophorous vacuole, while Pfg14.744 was detected in the erythrocyte. The 5' flanking region of either gene alone was sufficient to drive early gametocyte specific expression of green fluorescent protein (GFP). In parasites transfected with a plasmid containing the Pfg14.748 5' flanking region immediately upstream of GFP, fluorescence was observed in a small number of schizonts the cycle before stage I gametocytes were observed. This expression pattern is consistent with commitment to sexual differentiation prior to merozoite release and erythrocyte invasion. Further investigation into the role of these genes in the transition from asexual to sexual differentiation could provide new strategies to block malaria transmission. Microarray analysis was used to compare two clones derived from Plasmodium falciparum strain 3D7 parasites that differ in their ability to undergo gametocytogenesis. Clone G+ produces gametocytes and clone G- produces very few if any gametocytes. RNA was harvested from the cultures when the asexual parasitemia was 0.9-1.48% (day 4) (n=4) after setting up the gametocyte cultures and 5.2-5.58% (day 6) (n=4) prior to the appearance of morphologically distinct gametocytes and used to generate cDNA that was labeled with Cy3 or Cy5 and hybridized to the Plasmodium falciparum 70 mer oligonucleotide microarray developed by DeRisi and co-workers.

			Action	DRS
	EPIE_EN1_1.dat	Other
	EPIE_EN1_1.mgf	Mgf
	EPIE_EN1_1_100mM.mzXML	Mzxml
	EPIE_EN1_1_10mM.mzXML	Mzxml
	EPIE_EN1_1_15mM.mzXML	Mzxml