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Research Paper |
Research School of Biosciences, University of Kent, Canterbury, Kent CT2 7NJ, UK1
Centro de Genética e Biologia Molecular, Universidade de Lisboa, Portugal2
Author for correspondence: Mick F Tuite. Tel: +44 1227 823699. Fax: +44 1227 763912. e-mail: M. F.Tuite{at}ukc.ac.uk
The Sup35p protein of Saccharomyces cerevisiae is an essential translation factor whose prion-like properties give rise to the non-Mendelian genetic element [PSI+]. In this study the SUP35 gene from the related yeast species Candida albicans has been characterized. The CaSUP35 gene encodes a protein (CaSup35p) of 729 aa which shows 65% amino acid identity to the S. cerevisiae Sup35p protein (ScSup35p), with the C-terminal region showing greater identity (79%) than the N-terminal region. The full-length CaSup35p can functionally replace ScSup35p in S. cerevisiae although complementation is only complete when CaSup35p is overexpressed. Complementation only requires expression of the CaSup35p C domain. In S. cerevisiae the full-length CaSup35p is unable to establish a prion-like aggregated state even in the presence of endogenous ScSup35p prion seeds, thus confirming the existence of a species barrier in fungal prion propagation. Subcellular localization studies in C. albicans show that although CaSup35p is normally ribosome-associated, when not ribosome-associated, it does not form pelletable high-molecular-mass aggregates characteristic of the ScSup35p in [PSI+] strains. Unlike the ScSup35p, the CaSup35p N domain contains a number of polyglutamine repeats although it does contain seven copies of the peptide GGYQQ that is repeated in the ScSup35p N domain. Analysis of the CaSUP35 gene from 14 different strains of C. albicans identified four naturally occurring polymorphisms associated with changes in the length of the largest of the polyglutamine repeats. These findings have important implications for the evolution of fungal prion genes.
Keywords: SUP35 gene, polyQ, translation termination, [PSI+] determinant
The GenBank accession numbers for the sequences determined in this work are AF020554 (CaSUP35 gene) and AY028660AY028673 (CaSUP35 N domains).
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