ALTERNATIVE SPLICING: ANCESTRAL MECHANISM FOR THE HEXOSAMINIDASE SYNTHESIS?

MORALES, ENRIQUE SALVADOR; BANCHIO, CLAUDIA; KRAPF, DARÍO; ARRANZ, SILVIA

Chascomús

Congreso; XV ANNUAL MEETING OF THE ARGENTINE BIOLOGY SOCIETY (XV JORNADAS ANUALES DE LA SOCIEDAD ARGENTINA DE BIOLOGÍA); 2013

Sociedad Argentina de Biología

The N-ac-glucosaminidase (Hex) has been linked to several metabolic processes. In mammals, three Hex isoforms are originated by combinations of two different subunits (alfa and beta), encoded in two different genes. Their specific activities depend on the identity of two amino acids of the 12 that compose Hex active site. In our laboratory we characterized the putative amphibians Hex gene. Our results show that alfa and beta subunits can be generated from only one gen by alternative usage of 2 exons. One of the transcripts originated of this gene was cloned and sequenced (GenBank: JN127371; X.laevis). The deduced protein presented an active site corresponding to an alfa subunit. We aimed to express JN127371 and measure Hex activity in order to characterize its identity. JN127371 was subcloned in a pcDNA3 vector and transfected in CHO cells. Western-blot experiments demonstrated the expression of a protein of the expected molecular weight. Using substrates capable of discriminating between the alfa and beta subunit, a significant increase in alfa activity was observed in these extracts. Our results demonstrate that the characterized gene correspond to Hex and that JN127371 identifies as an alfa subunit, evidencing the possible common evolutionary origin of the alfa and beta Hex genes present in mammals.