Please use this identifier to cite or link to this item: http://hdl.handle.net/10261/40708
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Título : Functional Complementation in Yeast Allows Molecular Characterization of Missense Argininosuccinate Lyase Mutations
Autor : Trevisson, Eva, Burlina, Alberto, Doimo, Mara, Pertegato, Vanessa, Casarin, Alberto, Cesaro, Luca, Navas, Plácido, Basso, Giuseppe, Sartori, Geppo, Salviati, Leonardo
Palabras clave : Alleles
Argininosuccinate lyase
Genetic vectors
Genotype
Haploidy
Heterozygote
Phenotype
Saccharomyces cerevisiae
Urea
Fecha de publicación : 24-Aug-2009
Editor: American Society for Biochemistry and Molecular Biology
Citación : The Journal of Biological Chemistry 284(): 28926-28934 (2009)
Resumen: Deficiency of argininosuccinate lyase (ASL) causes argininosuccinic aciduria, an urea cycle defect that may present with a severe neonatal onset form or with a late onset phenotype. To date phenotype-genotype correlations are still not clear because biochemical assays of ASL activity correlate poorly with clinical severity in patients. We employed a yeast-based functional complementation assay to assess the pathogenicity of 12 missense ASL mutations, to establish genotype-phenotype correlations, and to screen for intragenic complementation. Rather than determining ASL enzyme activity directly, we have measured the growth rate in arginine-free medium of a yeast ASL(null) strain transformed with individual mutant ASL alleles. Individual haploid strains were also mated to obtain diploid, "compound heterozygous" yeast. We show that the late onset phenotypes arise in patients because they harbor individual alleles retaining high residual enzymatic activity or because of intragenic complementation among different mutated alleles. In these cases complementation occurs because in the hybrid tetrameric enzyme at least one active site without mutations can be formed or because the differently mutated alleles can stabilize each other, resulting in partial recovery of enzymatic activity. Functional complementation in yeast is simple and reproducible and allows the analysis of large numbers of mutant alleles. Moreover, it can be easily adapted for the analysis of mutations in other genes involved in urea cycle disorders.
Descripción : 9 páginas, 9 figuras, 2 tablas.
Versión del editor: http://dx.doi.org/10.1074/jbc.M109.050195
URI : http://hdl.handle.net/10261/40708
ISSN: 0021-9258
DOI: 10.1074/jbc.M109.050195
Appears in Collections:(CABD) Artículos

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