Trypanosoma cruzi bromodomain factor 2 (BDF2) binds to acetylated histones and is accumulated after UV irradiation

Villanova, Gabriela Vanina; Nardelli, Sheila Cristina; Cribb, Pamela; Magdaleno, Anahí; Silber, Ariel Mariano; Motta, María Cristina M.; Schenkman, Sérgio; Serra, Esteban

doi:10.1016/j.ijpara.2008.11.013

Cited by 42 publications

(47 citation statements)

References 37 publications

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“…6C). There was no cross-reactivity between bromodomain factors: recombinant TcBDF2 recognized only the H4K14Ac peptide (25), and TcBDF3 recognized only the acetylated ␣-tubulin peptide. This suggests that each BDF can recognize and bind to one (or a limited number of) specific acetylated lysine residues.…”

Section: Resultsmentioning

confidence: 97%

“…The TcBDF3-specific immunoreactive band was observed only in the nonnuclear fraction. TcTAT (39) is a cytosolic protein, and TcHMGB (30), TcBDF2 (25), and histone H4 (40) are all nuclear proteins (Fig. 1A).…”

Section: Resultsmentioning

confidence: 99%

“…Protein extracts (30 to 50 g per well) were separated by SDS-PAGE and transferred to nitrocellulose membranes. The transferred proteins were visualized with Ponceau S. The membranes were treated with 10% nonfat milk in PBS for 2 h and then incubated with specific antibodies diluted in PBS for 3 h. The antibodies used were polyclonal rabbit and mouse anti-TcBDF3, monoclonal mouse anti-acetylated ␣-tubulin antibody clone 6-11B-1 (Sigma-Aldrich), monoclonal mouse anti-trypanosome ␣-tubulin clone TAT-1 (a gift from K. Gull, University of Oxford, Oxford, England, United Kingdom), mouse anti-paraflagellar rod protein 2 (a gift from Ariel Silber, University of São Pablo, São Pablo, Brazil), mouse anti-T. cruzi histone H4 (a gift from Sergio Shenkman, Universidade Federal de São Paulo, São Paulo, Brazil), rabbit anti-T. cruzi high-mobility group B (TcHMGB) (30), and anti-T. cruzi bromodomain factor 2 (TcBDF2) (25). Bound antibodies were detected using peroxidase-labeled anti-mouse or anti-rabbit IgGs (GE Healthcare) and ECL Prime (GE Healthcare) according to the manufacturer's protocol.…”

Section: Methodsmentioning

confidence: 99%

“…(24). We previously characterized T. cruzi BDF2 (TcBDF2), which binds histone H4 (25). Here, we describe bromodomain factor 3 from T. cruzi (TcBDF3), the first exclusively nonnuclear bromodomain-containing protein reported so far.…”

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confidence: 98%

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Trypanosoma cruzi Bromodomain Factor 3 Binds Acetylated α-Tubulin and Concentrates in the Flagellum during Metacyclogenesis

et al. 2014

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cBromodomains are highly conserved acetyl-lysine binding domains found mainly in proteins associated with chromatin and nuclear acetyltransferases. The Trypanosoma cruzi genome encodes at least four bromodomain factors (TcBDFs). We describe here bromodomain factor 3 (TcBDF3), a bromodomain-containing protein localized in the cytoplasm. TcBDF3 cytolocalization was determined, using purified antibodies, by Western blot and immunofluorescence analyses in all life cycle stages of T. cruzi. In epimastigotes and amastigotes, it was detected in the cytoplasm, the flagellum, and the flagellar pocket, and in trypomastigotes only in the flagellum. Subcellular localization of TcBDF3 was also determined by digitonin extraction, ultrastructural immunocytochemistry, and expression of TcBDF3 fused to cyan fluorescent protein (CFP). Tubulin can acquire different posttranslational modifications, which modulate microtubule functions. Acetylated ␣-tubulin has been found in the axonemes of flagella and cilia, as well as in the subpellicular microtubules of trypanosomatids. TcBDF3 and acetylated ␣-tubulin partially colocalized in isolated cytoskeletons and flagella from T. cruzi epimastigotes and trypomastigotes. Interaction between the two proteins was confirmed by coimmunoprecipitation and far-Western blot assays with synthetic acetylated ␣-tubulin peptides and recombinant TcBDF3.

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Section: Resultsmentioning

confidence: 97%

Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

mentioning

confidence: 98%

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Trypanosoma cruzi Bromodomain Factor 3 Binds Acetylated α-Tubulin and Concentrates in the Flagellum during Metacyclogenesis

et al. 2014

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“…The first to be described was TcBDF2, which is expressed throughout the life cycle in the parasite nucleus and associates with chromatin through interactions with H4K10ac and H4K14ac. Little is known about the biological function of TcBDF2 other than that its levels increase following UV irradiation (88), so it may be involved in coordinating a response to DNA damage.…”

Section: Trypanosoma Cruzimentioning

confidence: 99%

Bromodomains in Protozoan Parasites: Evolution, Function, and Opportunities for Drug Development

Jeffers

Yang

Huang

et al. 2017

Microbiol Mol Biol Rev

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SUMMARY Parasitic infections remain one of the most pressing global health concerns of our day, affecting billions of people and producing unsustainable economic burdens. The rise of drug-resistant parasites has created an urgent need to study their biology in hopes of uncovering new potential drug targets. It has been established that disrupting gene expression by interfering with lysine acetylation is detrimental to survival of apicomplexan (Toxoplasma gondii and Plasmodium spp.) and kinetoplastid (Leishmania spp. and Trypanosoma spp.) parasites. As “readers” of lysine acetylation, bromodomain proteins have emerged as key gene expression regulators and a promising new class of drug target. Here we review recent studies that demonstrate the essential roles played by bromodomain-containing proteins in parasite viability, invasion, and stage switching and present work showing the efficacy of bromodomain inhibitors as novel antiparasitic agents. In addition, we performed a phylogenetic analysis of bromodomain proteins in representative pathogens, some of which possess unique features that may be specific to parasite processes and useful in future drug development.

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Distinct acetylation of Trypanosoma cruzi histone H4 during cell cycle, parasite differentiation, and after DNA damage

et al. 2009

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Histones of trypanosomes are quite divergent when compared to histones of most eukaryotes. Nevertheless, the histone H4 of Trypanosoma cruzi, the protozoan that causes Chagas' disease, is acetylated in the N terminus at lysines 4, 10, and 14. Here, we investigated the cellular distribution of histone H4 containing each one of these posttranslational modifications by using specific antibodies. Histone H4 acetylated at lysine 4 (H4-K4ac) is found in the entire nuclear space preferentially at dense chromatin regions, excluding the nucleolus of replicating epimastigote forms of the parasite. In contrast, histone H4 acetylated either at K10 or K14 is found at dispersed foci all over the nuclei and at the interface between dense and nondense chromatin areas as observed by ultrastructural immunocytochemistry. The level of acetylation at K4 decreases in nonreplicating forms of the parasites when compared to K10 and K14 acetylations. Antibodies recognizing the K14 acetylation strongly labeled cells at G2 and M stages of the cell cycle. Besides that, hydroxyurea synchronized parasites show an increased acetylation at K4, K10, and K14 after S phase. Moreover, we do not observed specific colocalization of K4 modifications with the major sites of RNA polymerase II. Upon γ-irradiation that stops parasite replication until the DNA is repaired, dense chromatin disappears and K4 acetylation decreases, while K10 and K14 acetylation increase. These results indicate that each lysine acetylation has a different role in T. cruzi. While K4 acetylation occurs preferentially in proliferating situations and accumulates in packed chromatin, K10 and K14 acetylations have a particular distribution probably at the boundaries between packed and unpacked chromatin.

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Trypanosoma cruzi bromodomain factor 2 (BDF2) binds to acetylated histones and is accumulated after UV irradiation

Cited by 42 publications

References 37 publications

Trypanosoma cruzi Bromodomain Factor 3 Binds Acetylated α-Tubulin and Concentrates in the Flagellum during Metacyclogenesis

Trypanosoma cruzi Bromodomain Factor 3 Binds Acetylated α-Tubulin and Concentrates in the Flagellum during Metacyclogenesis

Bromodomains in Protozoan Parasites: Evolution, Function, and Opportunities for Drug Development

Distinct acetylation of Trypanosoma cruzi histone H4 during cell cycle, parasite differentiation, and after DNA damage

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