Polymorphism of the yeast pyruvate carboxylase 2 gene and protein: Effects on biotinylation

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dc.contributor.authorVal, D.
dc.contributor.authorChapman-Smith, A.
dc.contributor.authorWalker, M.
dc.contributor.authorCronan, J.
dc.contributor.authorWallace, J.
dc.date.issued1995
dc.description.abstractIn Saccharomyces cerevisiae there are two isoenzymes of pyruvate carboxylase (Pyc) encoded by separate genes designated PYC1 and PYC2. We report the isolation and sequencing of a PYC2 gene, and the localization of both genes on the physical map of S. cerevisiae. Comparison with the previously reported sequence [Stucka, Dequin, Salmon and Gancedo (1991) Mol. Gen. Genet. 229, 307-315] revealed significant differences within the open reading frame. The most notable difference was near the 3' end, where we found a single base deletion reducing the open reading frame by 15 bases. We have confirmed the C-terminus of Pyc2 encoded by the gene isolated here by expressing and purifying an 86-amino-acid biotin-domain peptide. In addition, we investigated the effects of the two changes in the Pyc2 biotin domain (K1155R substitution and Q1178P/five-amino-acid extension) on the extent of biotinylation in vivo by Escherichia coli biotin ligase, and compared the biotinylation of peptides containing these changes with that of two different-length Pyc1 biotin-domain peptides. The K1155R substitution had very little effect on biotinylation, but the five-amino-acid C-terminal extension to Pyc2 and the N-terminal extension to Pycl both improved biotinylation in vivo.
dc.identifier.citationBiochemical Journal, 1995; 312(3):817-825
dc.identifier.doi10.1042/bj3120817
dc.identifier.issn0264-6021
dc.identifier.issn1470-8728
dc.identifier.orcidWalker, M. [0000-0002-6934-3787]
dc.identifier.urihttp://hdl.handle.net/2440/11269
dc.language.isoen
dc.publisherThe Biochemical Society, London
dc.source.urihttps://doi.org/10.1042/bj3120817
dc.subjectEscherichia coli
dc.subjectSaccharomyces cerevisiae
dc.subjectBiotin
dc.subjectIsoenzymes
dc.subjectPyruvate Carboxylase
dc.subjectDNA, Fungal
dc.subjectChromosome Mapping
dc.subjectSequence Analysis
dc.subjectMutagenesis
dc.subjectBinding Sites
dc.subjectAmino Acid Sequence
dc.subjectBase Sequence
dc.subjectSequence Homology
dc.subjectPolymorphism, Genetic
dc.subjectGenes, Fungal
dc.subjectMolecular Sequence Data
dc.titlePolymorphism of the yeast pyruvate carboxylase 2 gene and protein: Effects on biotinylation
dc.typeJournal article
pubs.publication-statusPublished

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