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However, in most cases binding of other TFs to the gene's promoter region determines the functionality of binding.
Our analysis suggests that the functionality of binding is highly condition-specific and highly dependent on the presence of specific cofactors.
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Factors that may determine the functionality of TF DNA binding are (i) distance of the binding site to the transcription start site (TSS), (ii) orientation of the TF with respect to the direction of transcription, (iii) local 3D structure of the DNA and (iv) presence or absence of interacting proteins (cofactors) in the same DNA region.
In this work, the influence of GU base pairs on the effectivity of translation attenuation by miRNAs is assayed by mutating binding sites and comparing attenuation effectivity to wild type binding sites Introducing three GU base pairs into the mRNA/miRNA duplex did, with only minor changes to the binding energy, almost completely destroy the functionality of the binding site.
Such competitive factors only affect the binding efficiency, but do not influence the functionality of the binding.
One way to cut through this noise and assess the functionality of TF binding is to leverage the effect of natural selection, which should tend to preserve functionally important binding events during evolution [ 6, 18- 20].
Finally, it is possible that other factors contribute to or modulate the functionality of PUM binding sites.
These findings further indicate that other factors may contribute to or modulate the functionality of PUM binding sites, for example recognition elements for cofactors like Nanos, which is known to interact with Pumilio to mediate translational repression [55].
Next, we employ the multi-parametric Random Forests machine learning technique to determine the factors controlling the functionality of TF binding.
To assess the functionality of σ binding sites defined by the ChIP-chip and PSSM analyses, promoter- lacZ fusion assays were performed using several of these sequences.
Some type of cooperativity among TFs, however, was an important component of the Gertz models and could explain the functionality of weaker binding sites, which have been argued to be prevalent in the yeast genome (Tanay 2006).
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