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Molecular Mechanisms of the Xanf1 Homeobox Gene Expression Regulation during the Early Development of the Forebrain Rudiment in the Clawed Frog

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Abstract

As a result of the analysis of the promoter region of the Xanf1 gene encoding the transcriptional regulator of the early development of the forebrain in the clawed frog, we found several promoter elements responsible for the expression of this gene and its localization in the anterior head region of the neural plate at the early neurula stage. A search for transcription factors that are able to bind to the found sites of the promoter was carried out. The binding of the found promoter elements to the transcription activators SoxD and Otx2 was shown using the electrophoretic mobility shift assay (EMSA) of DNA–protein complexes. We identified 10 inhibitory transcription factors, capable of binding to the regulatory element of the Xanf1 gene, using the procedure developed based on the yeast one-hybrid system. The effect of the found factors on the localization of the Xanf1 gene expression in the cells of the anterior head region of the neural plate was studied. In sum, the data allows us to put forward a hypothesis about the localization of the Xanf1 gene expression in the anterior head region of the neural plate, which completely corresponds to the classical Nieuwkoop model of neural induction.

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Abbreviations

MO:

morpholino oligonucleotides

BDGR:

binding domain of glucocorticoid receptor

EMSA:

electrophoretic mobility shift assay

FLD:

fluorescein lysine dextran

hd:

homeodomain

PMSF:

phenylmethylsulfonyl fluoride

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Correspondence to N. Y. Martynova.

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Original Russian Text © F.M. Eroshkin, A.V. Bayramov, G.V. Ermakova, A.G. Zaraisky, N.Y. Martynova, 2018, published in Bioorganicheskaya Khimiya, 2018, Vol. 44, No. 3, pp. 303–315.

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Eroshkin, F.M., Bayramov, A.V., Ermakova, G.V. et al. Molecular Mechanisms of the Xanf1 Homeobox Gene Expression Regulation during the Early Development of the Forebrain Rudiment in the Clawed Frog. Russ J Bioorg Chem 44, 310–321 (2018). https://doi.org/10.1134/S1068162018030032

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