Congresos y reuniones científicas
Título:
"The Palmitoyltramsferase Swf1 is a Bona Fide Zinc Binding Protein
Autor/es:
AYELEN GONZALEZ MONTORO; RODRIGO QUIROGA; JAVIER VALDEZ TAUBAS
Reunión:
Congreso; XLVIII SAIB; 2012
Institución organizadora:
Sociedad Argentina de Investigación en Bioquímica y Biología Molecular
Resumen:
S-acylation of proteins is catalysed by a family of
palmitoyltransferases characterised by the DHHC cysteine rich
domain. Swf1 is a member of this family, responsible for the
modification of SNAREs and glycosyltransferases in yeast. We
carried out a random mutagenesis assay designed to uncover
essential amino acids in Swf1 and identified 22 novel loss-offunction
mutations, most of which are localised within the DHHC
domain. Homology modelling of the tertiary structure of Swf1
DHHC-CRD shows that it could contain two C3H zinc binding
pockets, in a structure formed by three beta hairpins. The screen
revealed that mutation of each of the eight amino acids predicted to
be involved in zinc coordination results in inactive Swf1.All of these
mutations render Swf1 less stable, suggesting a structural role for
Swf1 zinc fingers. Sequence conservation studies of the amino acids
that form each zinc binding pocket indicate that upon mutation of
one of them the negative selection on the others is lost, in agreement
with a function which requires all four of them. Finally we show
directly that recombinant Swf1 DHHC-CRD domain is able to bind
zinc. Conservation and complementation studies suggest that the
DHHC domain of palmitoyltransferases which have lost the amino
acids that coordinate zinc may still acquire a similar structure by
alternative stabilization mechanisms.