COMPUTATIONAL AND PHENOTYPICAL COMPARISON BETWEEN THE RHOMBOID PROTEASES FROM Saccharomyces cerevisiae AND Escherichia coli

Rhomboid Proteases are part of a transmembrane protein superfamily involved in numerous relevant biological activities. This superfamily is divided into four groups: the group of serine proteases, which have catalytic activity; and other three groups that do not have catalytic activity. The functions and structural information of these proteins are studied, mainly, in E. coli, which makes it interesting to study the rhomboid proteases in eukaryotes, and allows to draw a comparative parallel observing the function, the structure and the evolution of this superfamily. Among the serine proteases (the rhomboids with catalytic activity) is the class of mitochondrial rhomboids that are found in all eukaryotes and located in the inner membrane of the mitochondria. In yeasts, which is the focus of this study, this enzyme is known as PCP1 (Processing of Cytochrome c peroxidase protein 1). Therefore, this study seeks, from computational and experimental data, to compare the rhomboid proteases in E. coli and S. cerevisiae. For comparison from computational data, the sequences were obtained in the Uniprot software, using the UniRef90 parameter to minimize redundancies and the alignments were made with the PFSTATS software. From these data was extracted information about the positions in which there is conservation of groups of residues, with structural and functional importance. In the comparison with experimental data, it was made the amplification of genes that encode the PCP1 protein via PCR, and subsequent transformation of yeasts and separation into groups that will be compared via spot test. The experimental groups will be formed by a wild type group (with the original protein) and mutant groups (with the protein with mutation regarding the residues of interest). The groups will also be subjected to different culture media in which the microorganism will perform respiration or fermentation in their metabolism. With these two approaches, it will be possible to observe if the phenotypic data corroborate with the structural data observed in silico and compare with the already published studies on E. coli, expanding the information on the superfamily of the rhomboids, so widely distributed.

This work was supported by the FAPEMIG Foundation, through a Master's Degree Scholarship.