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In agreement with prior data about condition-specific genes [ 8, 25], genes with high EV mainly have "covered" promoters with reduced histone acetylation and no RNA polymerase pre-initiation complex (PIC) occupancy, while genes with low EV have high PIC occupancy and increased histone acetylation in their promoters.
Overall, the computed free-energy landscape of nonconsensus protein-DNA binding shows strong correlation with the measured genome-wide PIC occupancy.
The second class (Class II) had PIC occupancy, however were not actively transcribed.
The first class (Class I) of genes had PIC occupancy and increased histone acetylation in their promoters and were actively transcribed.
We also show a correlation between HNF4A, TBP/TFIID, and PIC occupancy at distal sites, possibly enhancers, indicative of functional HNF4A TFIID interactions in vivo.
The third class of genes were actively transcribed although no PIC could be detected, while the fourth class (Class IV) had no PIC occupancy or detectable transcript levels and had reduced histone acetylation at their promoters [ 8].
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The genes from RNA-seq data (approx. 23K) were ranked relative to their expression level, divided into bins of 100, and attributed to the level of PIC component occupancy.
TAF4 regulates hepatocyte gene expression by controlling HNF4A occupancy, PIC formation, and Pol II pausing.
Mutual interactions between HNF4A and TFIID stabilise both PIC formation and CRM occupancy, without PIC formation HNF4A does not stably bind to its binding sites.
The concomitant loss of PIC formation and CRM occupancy upon TAF4 inactivation indicates that they are mutually dependent events.
We also noted that TAF3 and H3K4me3 levels did not fully correlate with Pol II occupancy and PIC formation at the most highly expressed genes.
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