A clustering method for robust and reliable large scale functional and structural protein sequence annotation

Bioinformatics, in the last few decades, has played a fundamental role to give sense to the huge amount of data produced. Obtained the complete sequence of a genome, the major problem of knowing as much as possible of its coding regions, is crucial. Protein sequence annotation is challenging and, due to the size of the problem, only computational approaches can provide a feasible solution. As it has been recently pointed out by the Critical Assessment of Function Annotations (CAFA), most accurate methods are those based on the transfer-by-homology approach and the most incisive contribution is given by cross-genome comparisons. In the present thesis it is described a non-hierarchical sequence clustering method for protein automatic large-scale annotation, called “The Bologna Annotation Resource Plus” (BAR+). The method is based on an all-against-all alignment of more than 13 millions protein sequences characterized by a very stringent metric. BAR+ can safely transfer functional features (Gene Ontology and Pfam terms) inside clusters by means of a statistical validation, even in the case of multi-domain proteins. Within BAR+ clusters it is also possible to transfer the three dimensional structure (when a template is available). This is possible by the way of cluster-specific HMM profiles that can be used to calculate reliable template-to-target alignments even in the case of distantly related proteins (sequence identity < 30%). Other BAR+ based applications have been developed during my doctorate including the prediction of Magnesium binding sites in human proteins, the ABC transporters superfamily classification and the functional prediction (GO terms) of the CAFA targets. Remarkably, in the CAFA assessment, BAR+ placed among the ten most accurate methods. At present, as a web server for the functional and structural protein sequence annotation, BAR+ is freely available at http://bar.biocomp.unibo.it/bar2.0.

Potential theory on metric spaces

This work revolves around potential theory in metric spaces, focusing on applications of dyadic potential theory to general problems associated to functional analysis and harmonic analysis. In the first part of this work we consider the weighted dual dyadic Hardy's inequality over dyadic trees and we use the Bellman function method to characterize the weights for which the inequality holds, and find the optimal constant for which our statement holds. We also show that our Bellman function is the solution to a stochastic optimal control problem. In the second part of this work we consider the problem of quasi-additivity formulas for the Riesz capacity in metric spaces and we prove formulas of quasi-additivity in the setting of the tree boundaries and in the setting of Ahlfors-regular spaces. We also consider a proper Harmonic extension to one additional variable of Riesz potentials of functions on a compact Ahlfors-regular space and we use our quasi-additivity formula to prove a result of tangential convergence of the harmonic extension of the Riesz potential up to an exceptional set of null measure