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Mol Biochem Parasitol. 2014 Oct;197(1-2):36-42. doi: 10.1016/j.molbiopara.2014.09.009. Epub 2014 Oct 5.

Canonical histone H2Ba and H2A.X dimerize in an opposite genomic localization to H2A.Z/H2B.Z dimers in Toxoplasma gondii.

Author information

1
Laboratorio de Parasitología Molecular, IIB-INTECH, CONICET-UNSAM, Av. Intendente Marino Km. 8.2, C.C 164, (B7130IIWA), Chascomús, Prov. Buenos Aires, Argentina.
2
Laboratorio de Parasitología Molecular, IIB-INTECH, CONICET-UNSAM, Av. Intendente Marino Km. 8.2, C.C 164, (B7130IIWA), Chascomús, Prov. Buenos Aires, Argentina; Scientific Research Commission (CIC, Buenos Aires), Argentina.
3
Department of Microbiology & Immunology, Albert Einstein College of Medicine, Bronx, NY, USA; Department of Pathology, Albert Einstein College of Medicine, Bronx, NY, USA; Department of Medicine, Albert Einstein College of Medicine, Bronx, NY, USA.
4
Department of Pharmacology & Toxicology, Indiana University School of Medicine, Indianapolis, IN, USA; Department of Microbiology & Immunology, Indiana University School of Medicine, Indianapolis, IN, USA.
5
Laboratorio de Parasitología Molecular, IIB-INTECH, CONICET-UNSAM, Av. Intendente Marino Km. 8.2, C.C 164, (B7130IIWA), Chascomús, Prov. Buenos Aires, Argentina. Electronic address: vanagas@intech.gov.ar.

Abstract

Histone H2Ba of Toxoplasma gondii was expressed as recombinant protein (rH2Ba) and used to generate antibody in mouse that is highly specific. Antibody recognizing rH2Ba detects a single band in tachyzoite lysate of the expected molecular weight (12kDa). By indirect immunofluorescence (IFA) in in vitro grown tachyzoites and bradyzoites, the signal was detected only in the parasite nucleus. The nucleosome composition of H2Ba was determined through co-immunoprecipitation assays. H2Ba was detected in the same immunocomplex as H2A.X, but not with H2A.Z. Through chromatin immunoprecipitation (ChIP) assays and qPCR, it was observed that H2Ba is preferentially located at promoters of inactive genes and silent regions, accompanying H2A.X and opposed to H2A.Z/H2B.Z dimers.

KEYWORDS:

Canonical histones; ChIP; Chromatin; Epigenetics; Nucleosome; Toxoplasma

PMID:
25286383
PMCID:
PMC4254167
DOI:
10.1016/j.molbiopara.2014.09.009
[Indexed for MEDLINE]
Free PMC Article

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