al independent branches of gap junction formation signaling pathways suggests an inverse causal relationship between the function of gap junctions and cholesterol homeostasis. Found at: doi:10.1371/journal.pone.0005197.s004 Acknowledgments We thank Justin Warner, John Alford, and Charles Tao for technical SU11274 manufacturer assistance with the genome-wide screen. 
Synonymous DNA variations may affect mRNA function through the change of mRNA secondary structure, mRNA stability, synonymous codon usage, or co-translational protein folding. With empirical evidence, synonymous single nucleotide polymorphisms in the COMT gene may modulate pain sensitivity through the effect on mRNA secondary structure and efficiency of protein expression. Examples of associations of sSNPs and human complex traits like the COMT sSNPs in pain sensitivity are rare. Most probably, although not functionally neutral, the functional effects of sSNPs are largely minor, while the minor effects are not readily identifiable by traditional genetic association study. SC usage bias is a widespread phenomenon across biological species. A sSNP changing codon usage may be expected to fine-tune translational efficiency based on the availability of rare tRNAs. According to the ramp model of mRNA translation, except the second codon, the first 50 codons of mRNAs tend to favor rarer codons and have slower speed of translation. This rampmechanism is important in determining translation efficiency, preventing ribosome congestion, and allowing proper co-translational folding of proteins. Based on the ramp theory, human sSNPs at ramp regions may confront selection pressure because of their functional effect on codon usage. To identify the translational effect of an individual SNP is difficult. Instead, we tried to identify the overall selection effect on sSNPs in human genome in this study. We investigated the incidences of sSNPs in the 3rd,50th codons vs. those in the remainder codons after the 51st codon. Methods Fourfold degenerate site sSNPs with AG or CT substitutions in human genome were extracted from the NCBI dbSNP database build 134. Altogether, 39,276 sSNPs in 12,568 genes were collected. All SNP alleles were corresponding to the nucleotides in coding sequences. Among these FFDS sSNPs, 20,122 were AG sSNPs, and 19,154 were CT sSNPs. Of the 20,122 AG FFDS sSNPs, 43 at second codons of coding regions were removed from further analysis; of 19,154 CT sSNPs, 25 at second codons were removed from further analysis. The FFDS sSNPs were annotated as N1RN2, while N1 represents the ancestral allele and N2 represents the variant allele. Ancestral alleles of sSNPs were inferred by human-chimpanzee genomic alignment according to the SeattleSeq Annotation 134. All sSNPs were differentiated by CpG sites versus non-CpG sites, while a CpG site has the pattern of YpG or CpR. 16483784 Results Our results showed that the fraction of FFDS sSNPs is significantly lower in the ramp than the Variant Direction Bias of 18772318 Synonymous SNPs rest regions . In the 3rd,50th codons, GRA sSNPs are favored than ARG sSNPs at non-CpG sites, and CRT sSNPs are favored than TRC sSNPs at non-CpG sites. In both cases of GRA and CRT, the favored direction of SC usage is the change from more frequent SCs to less frequent SCs. The reference data of human codon usage was calculated by the EMBL human coding sequences data release 115. By further investigation, our study disclosed that the GRA bias was mainly seen in synonymous substitution CG at non-CpG
