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The molecular function map for the EWS-FLI1 down-regulated genes shows two main clusters.
In order to construct a molecular function map of ESFT, information about EWS-FLI1 regulated genes was combined from two sources using the same microarray platform.
A molecular function map of EWS-FLI1 target genes was constructed by annotating the stringent set of EWS-FLI1 regulated genes defined above (GS1) using data bases of gene-function relationships.
When compared to the molecular function map developed in this study (Figure 3) these annotations overlap with only a subset of the molecular functions of EWS-FLI1 deregulated genes.
Out of 80 normal tissues tested, mesenchymal progenitor cells (MPC) were found to fit the hypothesis that EWS-FLI1 is the driving transcriptional force in ESFT best and were therefore used as the reference tissue for the construction of the molecular function map.
The molecular function map highlighted distinct clusters of activities for EWS-FLI1 regulated genes in ESFT and revealed a striking difference between EWS-FLI1 up- and down-regulated genes: EWS-FLI1 induced genes mainly belong to cell cycle regulation, proliferation, and response to DNA damage, while repressed genes were associated with differentiation and cell communication.
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This regional specialisation was also reflected among the molecular functions mapped to the enriched gene sets (Fig. 5).
Molecular-linkage maps based on functional gene markers (molecular-function maps) are a prerequisite for a candidate-gene approach to identify genes responsible for quantitative traits at the molecular level.
A total of 16,994 unique proteins mapped to 114,154 GO terms: 48,270 unigenes mapped to biological processes, 40,247 unigenes mapped to molecular functions, and 48,151 unigenes mapped to cellular components.EuKaryotic Orthologous Groups (KOG) classification of the unigenes is important for functional annotation and evolutionary studies.
Genes were categorized based on their molecular functions using the software, mapped onto genetic networks in the IPA database and then ranked by score.
It is widely accepted that most proteins exert their biological functions in part through interaction with other proteins, thus providing a rationale for efforts to infer molecular functions from protein protein interaction maps or 'interactomes' (von Mering et al., 2002).
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