TAILIEUCHUNG - Báo cáo khoa học: Organizational constraints on Ste12 cis-elements for a pheromone response in Saccharomyces cerevisiae

Ste12 ofSaccharomyces cerevisiaebinds to pheromone-responsecis-elements (PREs) to regulate several classes of genes. Genes induced by pheromones require multimerization of Ste12 for binding of at least two PREs on respon-sive promoters. We have systematically examined nucleotides of the consen-sus PRE for binding of wild-type Ste12 to DNA in vitro, as well as the organizational requirements of PREs to produce a pheromone response in vivo. | ỊFEBS Journal Organizational constraints on Ste12 cis-elements for a pheromone response in Saccharomyces cerevisiae Ting-Cheng Su1 2 Elena Tamarkina1 and Ivan Sadowski1 1 Department of Biochemistry and Molecular Biology Molecular Epigenetics LSI University of British Columbia Vancouver Canada 2 Graduate Program in Genetics University of British Columbia Vancouver Canada Keywords gene regulation pheromone response PRE Ste12 yeast Correspondence I. Sadowski Department of Biochemistry and Molecular Biology University of British Columbia 2350 Health Sciences Mall Vancouver BC V6T 1Z3 Canada Fax 1 604 822 9311 Tel 1 604 822 4524 E-mail sadowski@ Received 1 April 2010 revised 30 May 2010 accepted 3 June 2010 doi Ste12 of Saccharomyces cerevisiae binds to pheromone-response cis-elements PREs to regulate several classes of genes. Genes induced by pheromones require multimerization of Ste12 for binding of at least two PREs on responsive promoters. We have systematically examined nucleotides of the consensus PRE for binding of wild-type Ste12 to DNA in vitro as well as the organizational requirements of PREs to produce a pheromone response in vivo. Ste12 binds as a monomer to a single PRE in vitro and two PREs upstream of a minimal core promoter cause induction that is proportional to their relative affinity for Ste12 in vitro. Although consensus PREs are arranged in a variety of configurations in the promoters of responsive genes we find that there are severe constraints with respect to how they can be positioned in an artificial promoter to cause induction. Two closely-spaced PREs can induce transcription in a directly-repeated or tail-to-tail orientation although PREs separated by at least 40 nucleotides are capable of inducing transcription when oriented in a head-to-head or tail-to-tail configuration. We characterize several examples of promoters that bear multiple consensus PREs or a single PRE in combination with a .

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