A cis-acting element present in multiple genes serves as a repressor protein binding site for the yeast CAR1 gene.

Molecular and Cellular Biology
R M LucheT G Cooper

Abstract

Induction of the arginase (CAR1) gene expression in Saccharomyces cerevisiae has previously been shown to require participation of a cis-dominantly regulated upstream repression sequence (URS). Deletion of this element results in high-level expression of the CAR1 gene without inducer. To determine the structure of the CAR1 URS element, we performed a saturation mutagenesis. Results of the mutagenic analysis indicated that the CAR1 URS was a 9-base-pair palindromic sequence, 5'-AGCCGCCGA-3'. A DNA fragment containing this sequence was shown to bind one or more proteins by a gel shift assay. DNA fragments containing point mutations that completely eliminated URS function were not effective competitors in this assay, whereas those which supported URS function were effective competitors. Sequences in the 5'-flanking regions of 14 other genes were found to be homologous to the CAR1 URS. These sequences were shown to support varying degrees of URS function in the expression vector assay, to bind protein as demonstrated by the gel shift assay, and to compete with a DNA fragment containing the CAR1 URS for protein binding. These results indicate that the CAR1 URS element possesses the characteristics of a repressor binding site. Furthe...Continue Reading

References

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Dec 1, 1982·Molecular and Cellular Biology·R A Sumrada, T G Cooper

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Citations

Apr 22, 2004·Journal of Molecular Biology·Andreu Ferrer-MartínezAnna Maria Gómez-Foix
Dec 1, 1991·Yeast·R G Buckholz, T G Cooper
Mar 10, 1995·The Journal of Biological Chemistry·Y Hu, G B Kohlhaw
Mar 10, 1995·Molecular & General Genetics : MGG·J Bordallo, P Suárez-Rendueles
Jan 1, 1995·Molecular Microbiology·M KünzlerG H Braus
Mar 4, 2000·The Journal of Biological Chemistry·S ScottT G Cooper
Mar 10, 2005·Eukaryotic Cell·Mary E GardockiJohn M Lopes
Apr 10, 2020·European Journal of Immunology·Lorena Valverde-EstrellaAntonio Celada
Apr 17, 1999·Molecular and Cellular Biology·I IraquiA Urrestarazu
Mar 15, 1996·FEMS Microbiology Letters·C Martínez-CampaF Moreno
Jul 1, 1997·Molecular and Cellular Biology·V Gailus-DurnerA K Vershon
Mar 1, 1996·Microbiological Reviews·M L Greenberg, J M Lopes
Jun 8, 2021·Molecular Systems Biology·Yuko AritaR Scott McIsaac

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