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Furthermore, a sizable fraction of the measured differences can be attributed to divergent assignments of organism or splice isoforms, different organism focus and alternative representations of multi-protein complexes.
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We therefore calculated a corrected agreement measure for which we defined a range of translation scores around each group median where divergent group assignments were not counted.
Indeed, divergent organism assignments, the use of alternative protein identifiers, or different representation of complexes, although not reflecting actual curation errors, may lead to misinformation.
In order to factor out the effects of divergent organism assignments, we further restrict our analysis to co-citations in which both databases record proteins from the same organism.
In such co-citations improvements in annotation consistency of the order of 20% are observed for several organism categories (Table 1 and Figure 2c and d), confirming that divergent organism assignments contribute significantly to the observed differences.
Clearly the pairwise agreement levels for these organism-specific co-citations are significantly higher than those obtained for co-citations prior to factoring out divergent protein representation and/or organism assignments.
In order to determine how much expression signal is lost via reduced contig assignment when using divergent species as proxy references, expression values were calculated for GRS genes that are both represented in the de novo TA and have an RBH ortholog in the CCDS dataset.
Nielsen et al. (2012) evaluated a similar panel of SNPs for assignment success in Atlantic cod in the eastern Atlantic and concluded that eight of the most divergent loci were required for accurate assignment.
The main limitation is that since families can contain very divergent sequences with divergent functions, the GO assignments tend to be to high level terms to avoid incorrect annotations.
We then quantify how this initial level of agreement is globally impacted by factors such as divergent annotation of organisms, different splice isoform assignments and alternative representations of multi-protein complexes.
Since these sequences are quite divergent, it is difficult to assign a taxonomic identity to them in many cases and we caution the over-interpretation of taxonomic assignments at GAST values greater than 0.20.
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