Polymorphism of the yeast pyruvate carboxylase 2 gene and protein: effects on protein biotinylation
| Autor: | D.L. Val, John E. Cronan, Michelle E. Walker, John C. Wallace, Anne Chapman-Smith |
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| Rok vydání: | 1995 |
| Předmět: |
Sequence analysis
Genes Fungal Molecular Sequence Data Saccharomyces cerevisiae Biotin Sequence Homology Biochemistry Protein biotinylation chemistry.chemical_compound Escherichia coli Amino Acid Sequence DNA Fungal Molecular Biology Peptide sequence Pyruvate Carboxylase Binding Sites Polymorphism Genetic Base Sequence biology Chromosome Mapping Cell Biology biology.organism_classification Molecular biology Pyruvate carboxylase Isoenzymes Open reading frame chemistry Mutagenesis Biotinylation Sequence Analysis Research Article |
| Zdroj: | Biochemical Journal. 312:817-825 |
| ISSN: | 1470-8728 0264-6021 |
| DOI: | 10.1042/bj3120817 |
| Popis: | In 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. |
| Databáze: | OpenAIRE |
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