Polysome profiling reveals translational control of gene expression in the human malaria parasite Plasmodium falciparum

BackgroundIn eukaryotic organisms, gene expression is regulated at multiple levels during the processes of transcription and translation. The absence of a tight regulatory network for transcription in the human malaria parasite suggests that gene expression may largely be controlled at post-transcriptional and translational levels.ResultsIn this study, we compare steady-state mRNA and polysome-associated mRNA levels of Plasmodium falciparum at different time points during its asexual cell cycle. For more than 30% of its genes, we observe a delay in peak transcript abundance in the polysomal fraction as compared to the steady-state mRNA fraction, suggestive of strong translational control. Our data show that key regulatory mechanisms could include inhibitory activity of upstream open reading frames and translational repression of the major virulence gene family by intronic transcripts. In addition, we observe polysomal mRNA-specific alternative splicing events and widespread transcription of non-coding transcripts.ConclusionsThese different layers of translational regulation are likely to contribute to a complex network that controls gene expression in this eukaryotic pathogen. Disrupting the mechanisms involved in such translational control could provide novel anti-malarial strategies.

[1]  W. Trager,et al.  Human malaria parasites in continuous culture. , 1976, Science.

[2]  C. Lambros,et al.  Synchronization of Plasmodium falciparum erythrocytic stages in culture. , 1979, The Journal of parasitology.

[3]  M. Berry,et al.  Type I iodothyronine deiodinase is a selenocysteine-containing enzyme , 1991, Nature.

[4]  M. Kozak Structural features in eukaryotic mRNAs that modulate the initiation of translation. , 1991, The Journal of biological chemistry.

[5]  J. Yates,et al.  An approach to correlate tandem mass spectral data of peptides with amino acid sequences in a protein database , 1994, Journal of the American Society for Mass Spectrometry.

[6]  J. McCarthy,et al.  The Relationship between Eukaryotic Translation and mRNA Stability , 1995, The Journal of Biological Chemistry.

[7]  D. Bedwell,et al.  The efficiency of translation termination is determined by a synergistic interplay between upstream and downstream sequences in Saccharomyces cerevisiae. , 1995, Journal of molecular biology.

[8]  A. Geballe,et al.  Translational inhibition by a human cytomegalovirus upstream open reading frame despite inefficient utilization of its AUG codon , 1995, Journal of virology.

[9]  Mats Wahlgren,et al.  Developmental selection of var gene expression in Plasmodium falciparum , 1998, Nature.

[10]  T. Franklin,et al.  Inhibitors of protein biosynthesis , 1998 .

[11]  G. Mize,et al.  The Inhibitory Upstream Open Reading Frame from Mammalian S-Adenosylmethionine Decarboxylase mRNA Has a Strict Sequence Specificity in Critical Positions* , 1998, The Journal of Biological Chemistry.

[12]  Jean-Pierre Rousset,et al.  UAG readthrough in mammalian cells: Effect of upstream and downstream stop codon contexts reveal different signals , 2001, BMC Molecular Biology.

[13]  T. Franklin,et al.  Biochemistry and Molecular Biology of Antimicrobial Drug Action , 1998, Springer Netherlands.

[14]  Timothy J. Mitchison,et al.  Comparison of three directly coupled HPLC MS/MS strategies for identification of proteins from complex mixtures: single-dimension LC-MS/MS, 2-phase MudPIT, and 3-phase MudPIT , 2002 .

[15]  J. Yates,et al.  DTASelect and Contrast: tools for assembling and comparing protein identifications from shotgun proteomics. , 2002, Journal of proteome research.

[16]  Patricia De la Vega,et al.  Discovery of Gene Function by Expression Profiling of the Malaria Parasite Life Cycle , 2003, Science.

[17]  J. Derisi,et al.  The Transcriptome of the Intraerythrocytic Developmental Cycle of Plasmodium falciparum , 2003, PLoS biology.

[18]  L. Aravind,et al.  Plasmodium Biology Genomic Gleanings , 2003, Cell.

[19]  Yingyao Zhou,et al.  Global analysis of transcript and protein levels across the Plasmodium falciparum life cycle. , 2004, Genome research.

[20]  C. Ouzounis,et al.  Comparative genomics of transcriptional control in the human malaria parasite Plasmodium falciparum. , 2004, Genome research.

[21]  J. McCarthy,et al.  Reinitiation and recycling are distinct processes occurring downstream of translation termination in yeast. , 2004, Journal of molecular biology.

[22]  Ian A. Williams,et al.  Genome-wide prediction of stop codon readthrough during translation in the yeast Saccharomyces cerevisiae. , 2004, Nucleic acids research.

[23]  Jonathan Schug,et al.  Widespread distribution of antisense transcripts in the Plasmodium falciparum genome. , 2004, Molecular and biochemical parasitology.

[24]  M. Madan Babu,et al.  Discovery of the principal specific transcription factors of Apicomplexa and their implication for the evolution of the AP2-integrase DNA binding domains , 2005, Nucleic acids research.

[25]  Alisson M. Gontijo,et al.  Telomeric Heterochromatin Propagation and Histone Acetylation Control Mutually Exclusive Expression of Antigenic Variation Genes in Malaria Parasites , 2005, Cell.

[26]  John R Yates,et al.  A Comprehensive Survey of the Plasmodium Life Cycle by Genomic, Transcriptomic, and Proteomic Analyses , 2005, Science.

[27]  Manoj T. Duraisingh,et al.  Heterochromatin Silencing and Locus Repositioning Linked to Regulation of Virulence Genes in Plasmodium falciparum , 2005, Cell.

[28]  Allan Jacobson,et al.  Ribosome occupancy of the yeast CPA1 upstream open reading frame termination codon modulates nonsense-mediated mRNA decay. , 2005, Molecular cell.

[29]  Weltgesundheitsorganisation World malaria report , 2005 .

[30]  Michael P Washburn,et al.  Proteomic analysis by multidimensional protein identification technology. , 2006, Methods in molecular biology.

[31]  Neil Hall,et al.  Regulation of Sexual Development of Plasmodium by Translational Repression , 2006, Science.

[32]  Alisson M. Gontijo,et al.  5′ flanking region of var genes nucleate histone modification patterns linked to phenotypic inheritance of virulence traits in malaria parasites , 2007, Molecular microbiology.

[33]  Joseph L DeRisi,et al.  Whole-genome analysis of mRNA decay in Plasmodium falciparum reveals a global lengthening of mRNA half-life during the intra-erythrocytic development cycle , 2007, Genome Biology.

[34]  T. Wellems,et al.  Mechanisms underlying mutually exclusive expression of virulence genes by malaria parasites , 2007, EMBO reports.

[35]  M. Ares,et al.  Structural RNAs of known and unknown function identified in malaria parasites by comparative genomics and RNA analysis. , 2007, RNA.

[36]  B. Barrell,et al.  Genome-wide discovery and verification of novel structured RNAs in Plasmodium falciparum. , 2008, Genome research.

[37]  Zbynek Bozdech,et al.  Quantitative protein expression profiling reveals extensive post-transcriptional regulation and post-translational modifications in schizont-stage malaria parasites , 2008, Genome Biology.

[38]  Christian Epp,et al.  Chromatin associated sense and antisense noncoding RNAs are transcribed from the var gene family of virulence genes of the malaria parasite Plasmodium falciparum. , 2008, RNA.

[39]  V. Mootha,et al.  Upstream open reading frames cause widespread reduction of protein expression and are polymorphic among humans , 2009, Proceedings of the National Academy of Sciences.

[40]  Jose-Juan Lopez-Rubio,et al.  Genome-wide analysis of heterochromatin associates clonally variant gene regulation with perinuclear repressive centers in malaria parasites. , 2009, Cell host & microbe.

[41]  Matthew D. Young,et al.  Gene ontology analysis for RNA-seq: accounting for selection bias , 2010, Genome Biology.

[42]  D. Silhavy,et al.  Plant upstream ORFs can trigger nonsense-mediated mRNA decay in a size-dependent manner , 2009, Plant Molecular Biology.

[43]  Lior Pachter,et al.  Sequence Analysis , 2020, Definitions.

[44]  M. Gerstein,et al.  RNA-Seq: a revolutionary tool for transcriptomics , 2009, Nature Reviews Genetics.

[45]  Gonçalo R. Abecasis,et al.  The Sequence Alignment/Map format and SAMtools , 2009, Bioinform..

[46]  Anika Wiese-Klinkenberg,et al.  Sucrose Control of Translation Mediated by an Upstream Open Reading Frame-Encoded Peptide1[W][OA] , 2009, Plant Physiology.

[47]  S. Lonardi,et al.  Supplemental Material to : Nucleosome landscape and control of transcription in the human malaria parasite , 2009 .

[48]  Aaron R. Quinlan,et al.  Bioinformatics Applications Note Genome Analysis Bedtools: a Flexible Suite of Utilities for Comparing Genomic Features , 2022 .

[49]  E. Winzeler,et al.  The Plasmodium eukaryotic initiation factor-2α kinase IK2 controls the latency of sporozoites in the mosquito salivary glands , 2010, The Journal of experimental medicine.

[50]  N. Friedman,et al.  Strand-specific RNA sequencing reveals extensive regulated long antisense transcripts that are conserved across yeast species , 2010, Genome Biology.

[51]  M. Washburn,et al.  Refinements to label free proteome quantitation: how to deal with peptides shared by multiple proteins. , 2010, Analytical chemistry.

[52]  Samuel A. Assefa,et al.  New insights into the blood-stage transcriptome of Plasmodium falciparum using RNA-Seq , 2010, Molecular microbiology.

[53]  Samuel S. Shepard,et al.  Critical association of ncRNA with introns , 2010, Nucleic acids research.

[54]  M. Washburn,et al.  Unraveling the Ubiquitome of the Human Malaria Parasite* , 2011, The Journal of Biological Chemistry.

[55]  Nicholas T. Ingolia,et al.  Ribosome Profiling of Mouse Embryonic Stem Cells Reveals the Complexity and Dynamics of Mammalian Proteomes , 2011, Cell.

[56]  M. Selbach,et al.  Global quantification of mammalian gene expression control , 2011, Nature.

[57]  Sandrine Dudoit,et al.  GC-Content Normalization for RNA-Seq Data , 2011, BMC Bioinformatics.

[58]  Michael F. Lin,et al.  Evidence of abundant stop codon readthrough in Drosophila and other metazoa. , 2011, Genome research.

[59]  J. F. Atkins,et al.  Stimulation of stop codon readthrough: frequent presence of an extended 3′ RNA structural element , 2011, Nucleic acids research.

[60]  C. Nicchitta,et al.  Polysome profiling of the malaria parasite Plasmodium falciparum. , 2011, Molecular and biochemical parasitology.

[61]  J. Derisi,et al.  RNA-Seq analysis of splicing in Plasmodium falciparum uncovers new splice junctions, alternative splicing and splicing of antisense transcripts , 2011, Nucleic acids research.

[62]  S. Lonardi,et al.  Nucleosome occupancy at transcription start sites in the human malaria parasite: a hard-wired evolution of virulence? , 2011, Infection, genetics and evolution : journal of molecular epidemiology and evolutionary genetics in infectious diseases.

[63]  X. Su,et al.  Directional gene expression and antisense transcripts in sexual and asexual stages of Plasmodium falciparum , 2011, BMC Genomics.

[64]  Richard D Emes,et al.  Analysis of the spatial and temporal arrangement of transcripts over intergenic regions in the human malarial parasite Plasmodium falciparum , 2013, BMC Genomics.

[65]  M. Ahmad,et al.  Plasmodium falciparum DOZI, an RNA helicase interacts with eIF4E. , 2013, Gene.

[66]  V. Theodorou,et al.  GATA3 acts upstream of FOXA1 in mediating ESR1 binding by shaping enhancer accessibility , 2013, Genome research.

[67]  Johannes Griss,et al.  The Proteomics Identifications (PRIDE) database and associated tools: status in 2013 , 2012, Nucleic Acids Res..

[68]  Sol Katzman,et al.  Frac-seq reveals isoform-specific recruitment to polyribosomes , 2013, Genome research.

[69]  Joshua A. Arribere,et al.  Roles for transcript leaders in translation and mRNA decay revealed by transcript leader sequencing , 2013, Genome research.

[70]  K. L. Le Roch,et al.  An Apicoplast Localized Ubiquitylation System Is Required for the Import of Nuclear-encoded Plastid Proteins , 2013, PLoS pathogens.

[71]  L. Hurst,et al.  Positively Charged Residues Are the Major Determinants of Ribosomal Velocity , 2013, PLoS biology.

[72]  Imran Ullah,et al.  Functional analysis of the 5' untranslated region of the phosphoglutamase 2 transcript in Plasmodium falciparum. , 2013, Acta tropica.

[73]  E. Lai,et al.  Widespread and extensive lengthening of 3′ UTRs in the mammalian brain , 2013, Genome research.

[74]  K. Deitsch,et al.  A molecular switch in the efficiency of translation reinitiation controls expression of var2csa, a gene implicated in pregnancy‐associated malaria , 2013, Molecular microbiology.