Entamoeba histolytica: protein arginine transferase 1a methylates arginine residues and potentially modify the H4 histone

Borbolla-Vázquez, Jessica; Orozco, Esther; Betanzos, Abigail; Rodríguez, Mario A.

doi:10.1186/s13071-015-0820-7

Cited by 12 publications

(21 citation statements)

References 44 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Experimental evidence indicates that chromatin may also regulate gene expression in E. histolytica , as it has been demonstrated that E. histolytica DNA is organized into chromatin and that the nucleosome (the fundamental unit of chromatin) consists of four canonical histone dimers. In addition, E. histolytica produces proteins that acetylate, deacetylate and methylate histones [ 6 , 8 , 24 ], which are responsible for chromatin structure modifications. Post-translational modifications of histones and DNA methylation are two of the epigenetic mechanisms that have been best studied in eukaryotic organisms.…”

Section: Discussionmentioning

confidence: 99%

Identification of repressive and active epigenetic marks and nuclear bodies in Entamoeba histolytica

et al. 2016

View full text Add to dashboard Cite

BackgroundIn human hosts, Entamoeba histolytica cysts can develop into trophozoites, suggesting that the life cycle of this parasite are regulated by changes in gene expression. To date, some evidence has suggested that epigenetic mechanisms such as DNA methylation and histone modification are involved in the regulation of gene expression in Entamoeba. Some post–translational modifications (PTMs) at the N-terminus of E. histolytica’s histones have been reported experimentally, including tri-methylation in the lysine 4 of histone H3 (H3K4me3) and dimethylation in the lysine 27 of histone H3 (H3K27me2), dimethylation of arginine 3 (H4R3me2) and the indirect acetylation of histone H4 in the N-terminal region. However, it is not known which residues of histone H4 are subject to acetylation and/or methylation or where in the nucleus these epigenetic marks are located.MethodsHistones from trophozoites of E. histolytica were obtained and analyzed by LC-MS/MS. WB assays were performed using antibodies against epigenetic marks (acetylated lysines and methylated arginines). Immunofluorescence assays (IFA) were carried out to determine the distribution of PTMs and the localization of DNA methylation as a heterochromatin marker. Nuclear bodies such as the nucleolus were identified by using antibodies against fibrillarin and nucleolin and speckles by using anti-PRP6 antibody.ResultsSome new PTMs in histone H4 of E. histolytica, such as the acetylation of lysines 5, 8, 12 and 16 and the monomethylation of arginine 3, were identified by WB. IFA demonstrated that some marks are associated with transcriptional activity (such as acetylation and/or methylation) and that these marks are distributed throughout the E. histolytica nucleus. Staining with antibodies against anti-pan-acetylated lysine H4 histone and 5-methyl cytosine showed that the activation and transcriptional repression marks converge. Additionally, two nuclear bodies, the nucleolus and speckles, were identified in this parasite.ConclusionsThis study provides the first evidence that the nucleus of E. histolytica is not compartmentalized and contains two nuclear bodies, the nucleolus and speckles, the latter of which was not identified previously. The challenge is now to understand how these epigenetic marks and nuclear bodies work together to regulate gene expression in E. histolytica.Electronic supplementary materialThe online version of this article (doi:10.1186/s13071-016-1298-7) contains supplementary material, which is available to authorized users.

show abstract

Section: Discussionmentioning

confidence: 99%

Identification of repressive and active epigenetic marks and nuclear bodies in Entamoeba histolytica

et al. 2016

View full text Add to dashboard Cite

show abstract

“…To our knowledge, no PRMT substrates have been identified in Entamoeba spp., nor have any functional studies been performed. Recently, PRMT1a of E. histolytica was reported to be associated with gene expression regulation by dimethylation of H4R8 [ 27 ]. T. gondii CARM1 (PRMT homologue) resulted in decreased parasite viability due to the transient expression of enzymedead mutant TgCARM1 in tachyzoites [ 28 ].…”

Section: Discussionmentioning

confidence: 99%

Identification of Protein Arginine Methyltransferase 5 as a Regulator for Encystation of <i>Acanthamoeba</i>

et al. 2016

View full text Add to dashboard Cite

Encystation is an essential process for Acanthamoeba survival under nutrient-limiting conditions and exposure to drugs. The expression of several genes has been observed to increase or decrease during encystation. Epigenetic processes involved in regulation of gene expression have been shown to play a role in several pathogenic parasites. In the present study, we identified the protein arginine methyltransferase 5 (PRMT5), a known epigenetic regulator, in Acanthamoeba castellanii. PRMT5 of A. castellanii (AcPRMT5) contained domains found in S-adenosylmethionine-dependent methyltransferases and in PRMT5 arginine-N-methyltransferase. Expression levels of AcPRMT5 were increased during encystation of A. castellanii. The EGFP-PRMT5 fusion protein was mainly localized in the nucleus of trophozoites. A. castellanii transfected with siRNA designed against AcPRMT5 failed to form mature cysts. The findings of this study lead to a better understanding of epigenetic mechanisms behind the regulation of encystation in cyst-forming pathogenic protozoa.

show abstract

“…For western blot (WB) assays, membranes were stained with Ponceau S Red staining solution (Sigma-Aldrich) and incubated 1 h with 5% non-fat milk. Then were incubated overnight (ON) with α-TRF-like I (1:2,500), α-TRF-like III (1:1,000), α-EhKMT4 (1:1,000) (Borbolla-Vázquez et al, 2015 ); α-H3K27me3 (1:1,000, Cell Signaling Technology), α-H4K20me3 (1:1,000, Abcam), α-Lamin B1 (1:1,000, Abcam), α-actin (1:1,000, Sigma-Aldrich) or α-Myc (1:1,000, Cell Signaling Technology) antibodies and then, incubated for 2 h with the corresponding horseradish peroxidase (HRP) labeled secondary antibodies (1:8,000, Santa Cruz Biotechnology). Protein bands were detected and visualized by chemiluminescence on X-ray films.…”

Section: Methodsmentioning

confidence: 99%

Telomeric Repeat-Binding Factor Homologs in Entamoeba histolytica: New Clues for Telomeric Research

Rendón-Gandarilla

Álvarez-Hernández

Castañeda-Ortiz

et al. 2018

Front. Cell. Infect. Microbiol.

Self Cite

View full text Add to dashboard Cite

Telomeric Repeat Binding Factors (TRFs) are architectural nuclear proteins with critical roles in telomere-length regulation, chromosome end protection and, fusion prevention, DNA damage detection, and senescence regulation. Entamoeba histolytica, the parasite responsible of human amoebiasis, harbors three homologs of human TRFs, based on sequence similarities to their Myb DNA binding domain. These proteins were dubbed EhTRF-like I, II and III. In this work, we revealed that EhTRF-like I and II share similarity with human TRF1, while EhTRF-like III shares similarity with human TRF2 by in silico approach. The analysis of ehtrf-like genes showed they are expressed differentially under basal culture conditions. We also studied the cellular localization of EhTRF-like I and III proteins using subcellular fractionation and western blot assays. EhTRF-like I and III proteins were enriched in the nuclear fraction, but they were also present in the cytoplasm. Indirect immunofluorescence showed that these proteins were located at the nuclear periphery co-localizing with Lamin B1 and trimethylated H4K20, which is a characteristic mark of heterochromatic regions and telomeres. We found by transmission electron microscopy that EhTRF-like III was located in regions of more condensed chromatin. Finally, EMSA assays showed that EhTRF-like III forms specific DNA-protein complexes with telomeric related sequences. Our data suggested that EhTRF-like proteins play a role in the maintenance of the chromosome ends in this parasite.

show abstract

Entamoeba histolytica: protein arginine transferase 1a methylates arginine residues and potentially modify the H4 histone

Cited by 12 publications

References 44 publications

Identification of repressive and active epigenetic marks and nuclear bodies in Entamoeba histolytica

Identification of repressive and active epigenetic marks and nuclear bodies in Entamoeba histolytica

Identification of Protein Arginine Methyltransferase 5 as a Regulator for Encystation of <i>Acanthamoeba</i>

Telomeric Repeat-Binding Factor Homologs in Entamoeba histolytica: New Clues for Telomeric Research

Contact Info

Product

Resources

About