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The functional response of upstream DNA to dynamic supercoiling in vivo
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The functional response of upstream DNA to dynamic supercoiling in vivo
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Journal Article
The functional response of upstream DNA to dynamic supercoiling in vivo
2008
Overview
Because RNA polymerase is a powerful motor, transmission of transcription-generated forces might directly alter DNA structure, chromatin or gene activity in mammalian cells. Here we show that transcription-generated supercoils streaming dynamically from active promoters have considerable consequences for DNA structure and function in cells. Using a tamoxifen-activatable Cre recombinase to excise a test segment of chromatin positioned between divergently transcribed metallothionein-IIa promoters, we found the degree of dynamic supercoiling to increase as transcription intensified, and it was very sensitive to the specific arrangement of promoters and
cis
elements. Using psoralen as an
in vivo
probe confirmed that, during transcription, sufficient supercoiling is produced to enable transitions to conformations other than B-DNA in elements such as the human
MYC
far upstream element (FUSE), which in turn recruit structure-sensitive regulatory proteins, such as FUSE Binding Protein (FBP) and FBP-Interacting Repressor (FIR). These results indicate that mechanical stresses, constrained by architectural features of DNA and chromatin, may broadly contribute to gene regulation.
Publisher
Nature Publishing Group US,Nature Publishing Group
Subject
/ Biomedical and Life Sciences
/ Chromatin Immunoprecipitation
/ DNA
/ DNA Topoisomerases - metabolism
/ DNA, Superhelical - chemistry
/ DNA, Superhelical - metabolism
/ Gene Expression Regulation - physiology
/ Guanine Nucleotide Exchange Factors - metabolism
/ Humans
/ Mammals
/ Rho Guanine Nucleotide Exchange Factors
/ RNA-Binding Proteins - metabolism
