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Gene Synthesis> | … 1). Yeast Nap1 (Nap1) was purified from E. coli using 6× histidine-tag following the vendor's instruction (GenScript, L00223-10) and quantified with Bradford reagents (Biosciences, 786–012) (Supplementary Fig. 1). 2.2. DNA substrates … | Get A Quote |
Eukaryotes have evolved a specific strategy to package DNA. The nucleosome is a 147-base-pair DNA segment wrapped around histone core proteins that plays important roles regulating DNA-dependent biosynthesis and gene expression. Chromatin remodeling complexes (RSC, Remodel the Structure of Chromatin) hydrolyze ATP to perturb DNA-histone contacts, leading to nucleosome sliding and ejection. Here, we utilized tethered particle motion (TPM) experiments to investigate the mechanism of RSC-mediated nucleosome remodeling in detail. We observed ATP-dependent RSC-mediated DNA looping and nucleosome ejection along individual mononucleosomes and dinucleosomes. We found that nucleosome assembly protein 1 (Nap1) enha... More