The bZIP Dimer Localizes at DNA Full-Sites Where Each Basic Region Can Alternately Translocate and Bind to Subsites at the Half-Site

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• 作者：I-San Chan ; Taufik Al-Sarraj ; S. Hesam Shahravan ; Anna V. Fedorova ; Jumi A. Shin
• 刊名：Biochemistry
• 出版年：2012
• 出版时间：August 21, 2012
• 年：2012
• 卷：51
• 期：33
• 页码：6632-6643
• 全文大小：488K
• 年卷期：v.51,no.33(August 21, 2012)
• ISSN：1520-4995

文摘

Crystal structures of the GCN4 bZIP (basic region/leucine zipper) with the AP-1 or CRE site show how each GCN4 basic region binds to a 4 bp cognate half-site as a single DNA target; however, this may not always fully describe how bZIP proteins interact with their target sites. Previously, we showed that the GCN4 basic region interacts with all 5 bp in half-site TTGCG (termed 5H-LR) and that 5H-LR comprises two 4 bp subsites, TTGC and TGCG, which individually are also target sites of the basic region. In this work, we explore how the basic region interacts with 5H-LR when the bZIP dimer localizes to full-sites. Using AMBER molecular modeling, we simulated GCN4 bZIP complexes with full-sites containing 5H-LR to investigate in silico the interface between the basic region and 5H-LR. We also performed in vitro investigation of bZIP鈥揇NA interactions at a number of full-sites that contain 5H-LR versus either subsite: we analyzed results from DNase I footprinting and electrophoretic mobility shift assay (EMSA) and from EMSA titrations to quantify binding affinities. Our computational and experimental results together support a highly dynamic DNA-binding model: when a bZIP dimer localizes to its target full-site, the basic region can alternately recognize either subsite as a distinct target at 5H-LR and translocate between the subsites, potentially by sliding and hopping. This model provides added insights into how 伪-helical DNA-binding domains of transcription factors can localize to their gene regulatory sequences in vivo.