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We examined the validity of previous uses of sliding-window analysis to test for regions of a protein under selective constraint or positive selection [e.g.], [1], [4], [7], [8,14].
Sliding-window analysis has widely been used to uncover synonymous (silent, dS) and nonsynonymous (replacement, dN) rate variation along the protein sequence and to detect regions of a protein under selective constraint (indicated by dN<dS) or positive selection (indicated by dN>dS).
This is likely to be a conservative test for purifying selection because many synonymous sites are under selective constraint in S. cerevisiae (Zhou et al. 2010), and intergenic regions are likely to contain constrained sequences other than TFBSs (e.g., nucleosome positioning elements, noncoding RNAs etc).
Andolfatto (2005) has concluded that non-coding regions of Drosophila are under selective constraint and also subject to bouts of adaptive selection.
If the dN/dS rate ratio is <1, there is evidence for purifying (negative) selection, and the duplicated genes are thought to be under selective constraint.
Because our SVM classifies every genomic window as either evolving under selective constraint or under drift, we reasoned that regions experiencing positive selection might be classified as constrained, as positive selection reduces diversity at linked sites (Maynard Smith and Haigh 1974).
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Because synonymous sites are free from selection and nonsynonymous positions are under selective constraints, the observed pattern clearly points to the effects of purifying selection as predicted theoretically (Kimura 1983).
Although nonsynonymous sites and RNAs are well known to be under selective constraints, the present study reveals selection on the D-loop region of mitochondrial genomes.
The protein-coding genes in the mammalian mtDNA evolve under selective constraints and reveal codon position bias with the mean ratio of substitutions at 1st, 2nd, and 3rd codon positions equal to 2∶1∶5 [36].
In addition, the non-synonymous substitutions in most duplicates are under selective constraints (dN/ dS < 1).
There is no apparent reason why Cg-defh1 could be more under selective constraints than the other Cg-defs.
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