Caractérisation du site actif de la relaxase d’ICESt3 de Streptococcus thermophilus

Abstract : Horizontal transfer of genetic elements play a key role in evolution of bacterial genomes. Among these elements some are transferred by conjugation, especially integrative and conjugative elements (ICEs). The relaxase, which is an essential protein for these conjugative elements, is a single stranded endonuclease that initiate the DNA transfer. My internship objective is the active site characterization of ICESt3 relaxase, an ICE found in Streptococcus thermophilus, member of a large ICE family especially represented among firmicutes. This relaxase belongs to a new relaxase family (MOBT family) which have never been studied biochemically until now. Different residues of this relaxase active site have been mutated, that aresusceptible to be involved in protein activity, or indirectly in metallic cofactor coordination (an allmark for these endonucleases). The development of a relaxase activity test was carried out. Mutated proteins were produced and tested, allowing the identification of the residues involved in relaxase activity and in cation coordination.
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Master thesis
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http://docnum.univ-lorraine.fr/public/BUS_M_2016_MONTEIRO_PHILIPPE.pdf
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https://hal.univ-lorraine.fr/hal-01824834
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Philippe Monteiro. Caractérisation du site actif de la relaxase d’ICESt3 de Streptococcus thermophilus. Biotechnologies. 2016. ⟨hal-01824834⟩

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