Web implementation
The algorithm described for the identification of regulatory regions by comparative sequence analysis has been implemented as an intuitive and easy to use web service named ConSite [23]. The implementation allows for three analysis modes: first, alignment and conserved-site analysis of two orthologous genomic sequences applying one or more TF profiles; second, conserved site analysis on a submitted alignment, which allows users to generate alignments from their preferred tools and allows for the analysis of longer genomic sequences; and third, a single-sequence analysis tool. The single-sequence service is functionally comparable to the TESS system [24], but utilizes the
JASPAR profile collection [15]. Alignment submission accepts the de facto standard CLUSTALW format [25]. In all operating modes, users are allowed to submit a cDNA sequence to define exon locations. Users may also submit new matrix profiles of their own construction.
Results can be obtained in three distinct report formats. Graphical view (Figure 3a) displays an alignment overview and conservation plots with x-axis reference for each submitted sequence. Positions of conserved TFBSs are indicated above the plot. The transcription-factor labels are equipped with mouse-over function to display additional data (the name and structural class of the factor, and the absolute and relative site scores), and are hyperlinked to further information on the TF and its binding profile (Figure 3b). The popup windows provide data summaries, including a sequence logo (graphical representation of the specificity of the profile based on position-specific information content [26]) with the corresponding profile from the database. Alignment view (Figure 3c) provides a detailed overview of the detected potential TFBSs displayed on the sequence. The numbering indicates positions in the actual sequences, and the predicted TFBSs are marked. For convenience, a tabular output of detected sites with associated details is also provided in Table view.