doi: 10

doi: 10.1186/1471-2164-10-38. Download Physique?S2, TIF file, Splenopentin Acetate 0.4 MB mbo001162658sf2.tif (441K) GUID:?AECE27A9-0FAF-471E-B84C-AAAFF4C47517 Data Set?S1&#x000a0: Transcriptomes of the wild-type, strains. Three biological replicates of cRNA made from RNA harvested from wild-type, parasites (WT, KO, and Cm, respectively) were hybridized to an Affymetrix expression array and analyzed using GeneSpring software as detailed in Materials and Methods. Both natural hybridization and log2-transformed values are shown. Genes with significant expression differences with their normalized expression are listed. (The full list includes 3,781 genes by ANOVA with Benjamini-Hochberg correction for multiple testing [ 0.05].) The initial gene list was further filtered to identify those genes that showed 2-fold changes in expression compared to at least one other strain (639 genes with membership in either the G1 or S/M gene sets). There were differences in gene expression between all three strains tested. As the cell cycle phenotype was only evident in but not complemented parasites or wild-type parasites, we manually filtered the list of 639 genes to compile a list of 58 genes differentially expressed in the strain compared to both the wild-type and strains. Among these were candidate genes that could impact the assembly of the centrosome during parasite replication (Fig.?7; highlighted in yellow in the table). Download Data Set?S1, XLSX file, 0.9 MB mbo001162658sd1.xlsx (916K) GUID:?1AC796D1-5198-4203-92FB-2301E25999F4 ABSTRACT The arginine methyltransferase family (PRMT) has been implicated in a variety of cellular processes, including signal transduction, epigenetic regulation, and DNA repair pathways. PRMT1 is usually thought to be responsible for the majority of PRMT activity in mutants lacking PRMT1 (gene. parasites exhibit morphological defects during cell division and grow slowly, and this phenotype reverses in the complemented strain. Tagged PRMT1 localizes primarily in the cytoplasm with enrichment at the pericentriolar material, and the strain lacking PRMT1 is unable to segregate progeny accurately. Unlike wild-type and complemented parasites, parasites have abnormal daughter buds, perturbed centrosome stoichiometry, and loss of synchronous replication. Whole-genome expression profiling demonstrated differences in expression of cell-cycle-regulated genes in the strain relative to the complemented and parental wild-type strains, but Ginkgolide C these changes do not correlate with a specific block in cell cycle. Although PRMT1s primary biological function was previously proposed to be methylation of histones, our studies suggest that PRMT1 plays an important role within the centrosome to ensure the proper replication Ginkgolide C of the parasite. IMPORTANCE Apicomplexan parasites include several important pathogens, including by modifying histone methylation. Using genetic techniques, we show that disruption of PRMT1 Ginkgolide C affects the parasites ability to perform accurate cell division. Our studies uncover an unexpected role for arginine methylation in centrosome biology and regulation of parasite replication. INTRODUCTION is usually a category B biodefense priority pathogen. The parasite is usually widespread in its host range and geographical distribution, with an age-adjusted prevalence of 12.9% in the United States in 2009 2009 to 2010 (1). can differentiate from the rapidly replicating tachyzoite stage into a latent cyst form, the bradyzoite stage. Differentiation between parasite life cycle stages is usually accompanied by significant alterations in the expressed transcriptome, as well as remodeling of chromatin structure, a major mechanism by which the access to genomic DNA is restricted and regulated. Histone modifications in the Apicomplexa are proposed to act in concert with other putative epigenetic information carriers (histone variants and small RNAs) and ApiAP2 DNA sequence-specific transcription factors (2) to trigger the recruitment from the transcriptional equipment to particular genes. Methylation of arginine and lysine residues in histones can modulate gene manifestation positively or adversely (3), and both arginine and lysine methyltransferases are suggested to make a difference regulators of gene manifestation in (4). A thorough repertoire of arginine methylation equipment exists in weighed against.