| Literature DB >> 12051841 |
Veronica De Sanctis1, Sabrina La Terra, Alessandro Bianchi, David Shore, Luciano Burderi, Ernesto Di Mauro, Rodolfo Negri.
Abstract
We have analyzed in detail the structure of RAP1-UAS(RPG) complexes in Saccharomyces cerevisiae cells using multi-hit KMnO(4), UV and micrococcal nuclease high-resolution footprinting. Three copies of the Rap1 protein are bound to the promoter simultaneously in exponentially growing cells, as shown by KMnO(4) multi-hit footprinting analysis, causing extended and diagnostic changes in the DNA structure of the region containing the UAS(RPG). Amino acid starvation does not cause loss of Rap1p from the complex; however, in vivo UV-footprinting reveals the occurrence of structural modifications of the complex. Moreover, low-resolution micrococcal nuclease digestion shows that the chromatin of the entire region is devoid of positioned nucleosomes but is susceptible to changes in accessibility to the nuclease upon amino acid starvation. The implications of these results for the mechanism of Rap1p action are discussed. (c) 2002 Elsevier Science Ltd.Entities:
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Year: 2002 PMID： 12051841 DOI： 10.1016/S0022-2836(02)00110-9
Source DB: PubMed Journal: J Mol Biol ISSN： 0022-2836 Impact factor: 5.469