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. 2020 May 1;48(10):5426–5441. doi: 10.1093/nar/gkaa316

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© The Author(s) 2020. Published by Oxford University Press on behalf of Nucleic Acids Research.

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 8. — The importance of A/T-rich regions flanking Y8RE for Yap8 binding to the ACR3 promoter. (A) Alignment of confirmed (ScACR3, ScFRM2, KlACR2, KlACR3) and putative (SkAcr3, LfACR3, LnACR3, TmACR3) Y8RE promoter regions targeted by Yap8 orthologues from S. cerevisiae (Sc), S. kudriavzevii (Sk), K. lactis (Kl), L. fermentati (Lf), L. nothofagi (Ln) and T. microellipsoides (Tm). Putative interaction sites of unstructured N-tails of Yap8 are indicated by black bars. The sequence conservation logo of 13-bp Y8REs together with 6-bp flanking regions was determined by the WebLogo application (https://weblogo.berkeley.edu). (B) Binding of Yap8 protein to the ACR3 promoter variants as determined by EMSA. Purified GST-Yap8 protein was incubated with indicated biotin-labeled oligonucleotides corresponding to Y8RE-containing promoter fragments of ACR3 gene followed by electrophoresis. (C) Fluorescence anisotropy assays performed with purified GST-Yap8 and the FAM-labeled wild type and G/C-rich ACR3 promoter (ACR3-M3) fragments.