Our main interest is the characterization of the molecular mechanisms involved in the maintenance of genetic stability. We aim to advance in the characterization, diversity, and evolution of the studied processes as well as in their possible biotechnological applications. We use multidisciplinary approaches, combining bioinformatics, biochemistry, molecular biology, and microbiology.
We are particularly interested in the biochemical characterization of enzymes involved in DNA replication and in alternative mechanisms of replication initiation or priming, independent of traditional DNA primases. For this aim, we work with different simple models such as viruses or bacterial mobile genetic elements.
Our current efforts are focused on family B DNA polymerases, especially the piPolBs ("primer-independent PolBs") subfamily, described and characterized by our laboratory. The piPolBs are probably at the origin of this family of DNA polymerases and show unexpected properties, such as the ability to initiate replication "de novo", without the need for a pre-existing primer. In the last years, we have focused on structure-function studies towards the dissection of the molecular mechanisms of each of the piPolB activities.
The piPolB features suggest a straightforward application on isothermal DNA amplification. Thus, we have developed a novel method for whole (meta)genome DNA amplification, with multiple applications on biotechnology and biomedicine.
The piPolBs are encoded in genetic mobile elements called pipolins, which are present in bacteria and in some mitochondria. Pipolins are not very frequent, but they are very diverse elements, whose only common factor is the presence of the gene that codes for a piPolB and they can be found both integrated in the genomes of various pathogenic bacteria, such as Escherichia coli or Vibrio parahaemolyticus, as well as in the form of plasmids, as is the case with Staphylococcus sp. or mitochondrial elements. Currently, we are working to understand the biological and biomedical relevance of pipolins at various levels. On the one hand, we use phylogenetic methods and network analysis to study their diversity and evolution. At a more specific level, we analyzed the prevalence of pipolins in clinical isolates and are also conducting genetic analyzes to reveal the function of piPolB and other E. coli pipolin genes in vivo.