FATTY ACID 2-HIDROXYLASE GENE IDENTIFICATION IN Tetrahymena thermophila MACRONUCLEAR GENOME

CID NICOLAS; MARIA LUZ SANCHEZ; CLARA NUDEL; ALEJANDRO D. NUSBLAT

Mar del Plata

Congreso; LI Reunión SAIB 201 5 ,; 2015

SAIB

Sphingolipids are lipids of great importance not only because of their structural function on cellular membranes, but also they are bioactive molecules, since them participate in several process as cellular signaling, vesicular traffic and apoptosis. Ciliates, as Tetrahymena thermophila, are promising organism to apply to the field of sphingolipid research, since they have many advantages in their use, including complex biological processes, ease of cultivation and various molecular tools. However, although it has been reported the presence of these lipids in Tetrahymena, neither their biological role, nor the genes involved in metabolism, are known. Addressing these issues, it has been built by bioinformatics searches a putative metabolic pathway in the ciliated, identifying possible paralogs for most of the synthetic steps involved. According to this, we proceeded to knock the only candidate of a fatty acid hydroxylase gene, transforming the macronucleus by biolistic bombardment. Results showed the loss of hydroxylase activity by gas chromatography without observing other significant phenotypic changes from the wild strain. In conclusion, Identifying fatty acid 2-hydroxylase enzyme´s gene thus becomes the first gene of sphingolipid metabolism to be identified, not only in T. thermophila, but generally in ciliates.