Destabilization of nucleosomes by an unusual DNA conformation adopted by poly(dA) small middle dotpoly(dT) tracts in vivo.
Poly(dA) small middle dotpoly(dT) tracts are common and often found upstream of genes in eukaryotes. It has been suggested that poly(dA) small middle dotpoly(dT) promotes transcription in vivo by affecting nucleosome formation. On the other hand, in vitro studies show that poly(dA) small middle dotpoly(dT) can be easily incorporated into nucleosomes. Therefore, the roles of these tracts in nucleosome organization in vivo remain to be established. We have developed an assay system that can evaluate nucleosome formation in yeast cells, and demonstrated that relatively longer tracts such as A(15)TATA(16) and A(34) disrupt an array of positioned nucleosomes, whereas a shorter A(5)TATA(4) tract is incorporated in positioned nucleosomes of yeast minichromosomes. Thus, nucleosomes are destabilized by poly(dA) small middle dotpoly(dT) in vivo in a length-dependent manner. Furthermore, in vivo UV footprinting revealed that the longer tracts adopt an unusual DNA structure in yeast cells that corresponds to the B' conformation described in vitro. Our results support a mechanism in which a unique poly(dA) small middle dot poly(dT) conformation presets chromatin structure to which transcription factors are accessible.
Shimizu
M
M
Department of Chemistry, Meisei University, Hino, Tokyo 191-8506 and School of Pharmacy, Tokyo University of Pharmacy and Life Science, Hachioji, Tokyo 192-0392, Japan. shimizum@chem.meisei-u.ac.jp
Mori
T
T
Sakurai
T
T
Shindo
H
H
D001483:Base Sequence; D018983:DNA Footprinting / Q000379:methods; D004271:DNA, Fungal / Q000737:chemistry*; D006868:Hydrolysis; D008836:Micrococcal Nuclease / Q000378:metabolism; D009690:Nucleic Acid Conformation; D009707:Nucleosomes / Q000737:chemistry* / Q000378:metabolism; D011067:Poly dA-dT / Q000737:chemistry*; D012441:Saccharomyces cerevisiae / Q000235:genetics; D014466:Ultraviolet Rays
D004271:DNA, Fungal; D009707:Nucleosomes; D011067:Poly dA-dT; D008836:Micrococcal Nuclease