TY - JOUR
T1 - Analysis of the mechanism of nucleosome survival during transcription
AU - Chang, Han Wen
AU - Kulaeva, Olga I.
AU - Shaytan, Alexey K.
AU - Kibanov, Mikhail
AU - Kuznedelov, Konstantin
AU - Severinov, Konstantin V.
AU - Kirpichnikov, Mikhail P.
AU - Clark, David J.
AU - Studitsky, Vasily M.
PY - 2014/2
Y1 - 2014/2
N2 - Maintenance of nucleosomal structure in the cell nuclei is essential for cell viability, regulation of gene expression and normal aging. Our previous data identified a key intermediate (a small intranucleosomal DNA loop, Ø-loop) that is likely required for nucleosome survival during transcription by RNA polymerase II (Pol II) through chromatin, and suggested that strong nucleosomal pausing guarantees efficient nucleosome survival. To evaluate these predictions, we analysed transcription through a nucleosome by different, structurally related RNA polymerases and mutant yeast Pol II having different histone-interacting surfaces that presumably stabilize the Ø-loop. The height of the nucleosomal barrier to transcription and efficiency of nucleosome survival correlate with the net negative charges of the histone-interacting surfaces. Molecular modeling and analysis of Pol II-nucleosome intermediates by DNase I footprinting suggest that efficient Ø-loop formation and nucleosome survival are mediated by electrostatic interactions between the largest subunit of Pol II and core histones.
AB - Maintenance of nucleosomal structure in the cell nuclei is essential for cell viability, regulation of gene expression and normal aging. Our previous data identified a key intermediate (a small intranucleosomal DNA loop, Ø-loop) that is likely required for nucleosome survival during transcription by RNA polymerase II (Pol II) through chromatin, and suggested that strong nucleosomal pausing guarantees efficient nucleosome survival. To evaluate these predictions, we analysed transcription through a nucleosome by different, structurally related RNA polymerases and mutant yeast Pol II having different histone-interacting surfaces that presumably stabilize the Ø-loop. The height of the nucleosomal barrier to transcription and efficiency of nucleosome survival correlate with the net negative charges of the histone-interacting surfaces. Molecular modeling and analysis of Pol II-nucleosome intermediates by DNase I footprinting suggest that efficient Ø-loop formation and nucleosome survival are mediated by electrostatic interactions between the largest subunit of Pol II and core histones.
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U2 - 10.1093/nar/gkt1120
DO - 10.1093/nar/gkt1120
M3 - Article
C2 - 24234452
SN - 0305-1048
VL - 42
SP - 1619
EP - 1627
JO - Nucleic Acids Research
JF - Nucleic Acids Research
IS - 3
ER -