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Engineering of monotransregulators.

By Mesut Muyan (785672), Gizem Güpür (785673), Pelin Yaşar (785674), Gamze Ayaz (785675), Sırma Damla User (785676), Hasan Hüseyin Kazan (785677) and Yanfang Huang (785678)


<p>(<b>A</b>) Schematic of ERα. 595 amino-acid long ERα is a modular protein that contains an amino-terminally located ligand-independent activation function, A/B, domain followed by the DNA binding, C, domain. The hinge, D, domain encompasses a nuclear localization signal and connects the C domain to the carboxyl-terminus, E/F, domain. E/F is a multi-functional domain containing ligand binding, dimerization and ligand-dependent activation functions. ERα binds to specific DNA sequences, so-called estrogen responsive elements (EREs), derivatives of the consensus GGTCAnnnTGACC wherein ‘n’ denotes three non-specific nucleotides. (<b>B</b>) The engineered monomeric ERE binding module CDC is composed of two C domains joined with the D domain. (<b>C</b>) The cDNA of the activation domain (AD) of p65 and VP16 proteins or of the repression domain (RD) KRAB of KOX-1 of and/or SID of Mad1 was genetically joined to the 5′ and 3′ ends, respectively, of the CDC-cDNA or the ERE binding defective CDC<sub>EBD</sub> to generate the monotransregulators. The constructs also contain sequences encoding the Flag and 6xHis epitopes at the amino and carboxyl-termini, respectively.</p

Topics: Uncategorised, rd, gene expressions, model monotransrepressor, estrogen response elements, dbd, DNA binding domain, transcription factors, er, monomeric transcription activators, chromatin context, Cell proliferation, protein engineering platform, Cellular Proliferation, heterologous repression domains, reporter gene expressions, Cell growth, ere, DNA synthesis, ad, Polar Directions, transcription activation domains, monotransactivator, ebm, cell cycle progression
Year: 2015
DOI identifier: 10.1371/journal.pone.0136423.g001
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Provided by: FigShare
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