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

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Abstract

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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Acknowledgments

We thank Dr. James Bangs (The University of Wisconsin) for the use of the three-dimensional deconvolution microscope and Claudio Rogerio de Oliveira for the technical help. This work was supported by grants from FAPESP, FAPERJ, and CNPq (Brazil). SCN and JPCC are FAPESP (Brazil) fellows.

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  1. Julia Pinheiro Chagas da Cunha

    Present address: Ludwig Institute for Cancer Research, Rua João Julião 245—Paraiso, São Paulo, São Paulo, 01323-903, Brazil

Authors and Affiliations

  1. Departamento de Microbiologia, Imunologia e Parasitologia, Universidade Federal de São Paulo, Rua Botucatu 862 8A, São Paulo, São Paulo, 04023-062, Brazil

    Sheila Cristina Nardelli, Julia Pinheiro Chagas da Cunha & Sergio Schenkman

  2. Instituto de Biofísica Carlos Chagas Filho, Universidade Federal do Rio de Janeiro, CCS, Bloco G, Ilha do Fundão, Rio de Janeiro, 21949-900, Rio de Janeiro, Brazil

    Maria Cristina M. Motta

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  1. Sheila Cristina Nardelli
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  4. Sergio Schenkman
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Correspondence to Sergio Schenkman.

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Communicated by E.A. Nigg

Sheila Cristina Nardelli and Julia Pinheiro Chagas da Cunha contributed equally to this work.

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Nardelli, S.C., da Cunha, J.P.C., Motta, M.C.M. et al. Distinct acetylation of Trypanosoma cruzi histone H4 during cell cycle, parasite differentiation, and after DNA damage. Chromosoma 118, 487–499 (2009). https://doi.org/10.1007/s00412-009-0213-9

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