Molecular characterization of the Schistosoma mansoni zinc finger protein SmZF1 as a transcription factor

Drummond, Marcela G., Calzavara-Silva, Carlos E., D'Astolfo, Diego S., Cardoso, Fernanda C., Rajao, Matheus A., Mourão, Marina M., Gava, Elisandra, Oliveira, Sérgio C., Macedo, Andréa M., Machado, Carlos R., Pena, Sérgio D. J., Kitten, Gregory T. and Franco, Glória R. (2009) Molecular characterization of the Schistosoma mansoni zinc finger protein SmZF1 as a transcription factor. PLoS Neglected Tropical Diseases, 3 11: e547.1-e547.10. doi:10.1371/journal.pntd.0000547

Author Drummond, Marcela G.
Calzavara-Silva, Carlos E.
D'Astolfo, Diego S.
Cardoso, Fernanda C.
Rajao, Matheus A.
Mourão, Marina M.
Gava, Elisandra
Oliveira, Sérgio C.
Macedo, Andréa M.
Machado, Carlos R.
Pena, Sérgio D. J.
Kitten, Gregory T.
Franco, Glória R.
Title Molecular characterization of the Schistosoma mansoni zinc finger protein SmZF1 as a transcription factor
Formatted title
Molecular characterization of the Schistosoma mansoni zinc finger protein SmZF1 as a transcription factor
Journal name PLoS Neglected Tropical Diseases   Check publisher's open access policy
ISSN 1935-2727
Publication date 2009-11
Sub-type Article (original research)
DOI 10.1371/journal.pntd.0000547
Open Access Status DOI
Volume 3
Issue 11
Start page e547.1
End page e547.10
Total pages 10
Place of publication San Francisco, CA, U.S.A.
Publisher Public Library of Science
Language eng
Formatted abstract
Background: During its development, the parasite Schistosoma mansoni is exposed to different environments and undergoes many morphological and physiological transformations as a result of profound changes in gene expression. Characterization of proteins involved in the regulation of these processes is of importance for the understanding of schistosome biology. Proteins containing zinc finger motifs usually participate in regulatory processes and are considered the major class of transcription factors in eukaryotes. It has already been shown, by EMSA (Eletrophoretic Mobility Shift Assay), that SmZF1, a S. mansoni zinc finger (ZF) protein, specifically binds both DNA and RNA oligonucleotides. This suggests that this protein might act as a transcription factor in the parasite.

Methodology/Principal Findings: In this study we extended the characterization of SmZF1 by determining its subcellular localization and by verifying its ability to regulate gene transcription. We performed immunohistochemistry assays using adult male and female worms, cercariae and schistosomula to analyze the distribution pattern of SmZF1 and verified that the protein is mainly detected in the cells nuclei of all tested life cycle stages except for adult female worms. Also, SmZF1 was heterologously expressed in mammalian COS-7 cells to produce the recombinant protein YFP-SmZF1, which was mainly detected in the nucleus of the cells by confocal microscopy and Western blot assays. To evaluate the ability of this protein to regulate gene transcription, cells expressing YFP-SmZF1 were tested in a luciferase reporter system. In this system, the luciferase gene is downstream of a minimal promoter, upstream of which a DNA region containing four copies of the SmZF1 putative best binding site (D1-3DNA) was inserted. SmZF1 increased the reporter gene transcription by two fold (p≤0.003) only when its specific binding site was present.

Conclusion: Taken together, these results strongly support the hypothesis that SmZF1 acts as a transcription factor in S. mansoni.

Author Summary: Schistosomes are parasites that exhibit a complex life cycle during which they progress through many morphological and physiological transformations. These transformations are likely accompanied by alterations in gene expression, making genetic regulation important for parasite development. Here we describe a Schistosoma mansoni protein (SmZF1) that may act as a parasite transcription factor. These factors are key proteins for gene regulation. We have previously demonstrated that SmZF1 is able to bind DNA and that its mRNA is present at different stages during the parasite life cycle. In this study we aimed to define if this protein can function as a transcription factor in S. mansoni. SmZF1 was detected in the nucleus of adult male worms, cercariae and schistosomula cells. It was not, however, observed in female cells, suggesting it to be gender specific. We used mammalian cells expressing recombinant SmZF1 to analyze if SmZF1 protein is able to activate/repress gene transcription and demonstrated that it increased the expression of a reporter gene by two-fold. The results obtained confirm SmZF1 as a S. mansoni transcription factor.
Q-Index Code C1
Q-Index Status Provisional Code
Institutional Status Non-UQ
Additional Notes Article # e547

Document type: Journal Article
Sub-type: Article (original research)
Collections: ERA 2012 Admin Only
Institute for Molecular Bioscience - Publications
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Citation counts: TR Web of Science Citation Count  Cited 3 times in Thomson Reuters Web of Science Article | Citations
Scopus Citation Count Cited 5 times in Scopus Article | Citations
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Created: Tue, 22 Mar 2011, 11:44:42 EST by Susan Allen on behalf of Institute for Molecular Bioscience