TRANSCRIPTION THROUGH NUCLEOSOMES BY RNA POLYMERASE II

Basic Sciences

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Abstract

The DNA within the eukaryotic nucleus is packaged as chromatin - a tight nucleoprotein complex composed of repeating units called nucleosomes. Pol II has developed a special mechanism to transcribe through such a tightly packed DNA. This project is aimed to investigate transcription by yeast Pol II elongation complexes through uniquely positioned nucleosome core in vitro. We apply recently developed experimental system employing assembly of authentic Pol II elongation complexes and ligation of these complexes to the mononucleosome in vitro. It has been found that this system faithfully recapitulates chief properties of the promoter-dependent experimental system such as the height and general properties of the nucleosomal barrier to transcription. We demonstrate that Pol II uses unique mechanism for transcription through chromatin. Surprising landmark of this mechanism is ability of the polymerase to transcribe through the nucleosome without accompanying change in nucleosome position observed in the case of all other analyzed RNA polymerases.