Abstract
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 nuclear proteins covering all expected nuclear protein families involved in gene regulation. All structural ribosomal (40S and 60S) proteins were identified which is consistent with the nuclear localization of ribosomal biogenesis. Proteins involved in the translation machinery were also found suggesting that translational events might occur in the nucleus in P. falciparum as previously hypothesized in eukaryotes. These data were compared to the protein list established by PlasmoDB and submitted to Plasmobase a recently reported Plasmodium annotation website to propose new functional putative annotation of several unknown proteins found in the nuclear extracts.
Highlights
In eukaryote cells, the nucleus is a highly dynamic and complex organelle [1] [2] where major regulatory gene expression events take place such as DNA replication, RNA synthesis within transcriptional machinery, mRNA processing and transport to the cytoplasm as well as ribosomal sub-units biogenesis
Our aim was to identify P. falciparum nuclear proteins with a special focus on all proteins involved in overall gene regulation from DNA replication to RNA transcription [23], mRNA maturation and transport as well as mRNA translation
P. falciparum nuclear protein identification and annotation validate the quality of our nuclear extracts, the content of nuclear (NE1 and NE2) and corresponding cytosolic extracts (CE1 and CE2) was evaluated by SDS-PAGE
Summary
The nucleus is a highly dynamic and complex organelle [1] [2] where major regulatory gene expression events take place such as DNA replication, RNA synthesis within transcriptional machinery, mRNA processing and transport to the cytoplasm as well as ribosomal sub-units biogenesis. The nucleus is organized to participate in RNA, protein and ribosomal sub-unit trafficking in and out of the nucleus [3]. A continuous cytoplasm-nucleus protein shuttling occurs in eukaryotes weakening the characterisation and definition of nuclear proteins even though proteins accumulate either in the cytoplasm or in the nucleus according to the cellular development [5]. This back and forth protein transport through the nuclear pore plays an important role in the control of gene expression
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