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Related Experiment Videos

MAT alpha 1 can mediate gene activation by a-mating factor.

P Sengupta1, B H Cochran

  • 1MIT Center for Cancer Research and Department of Biology, Cambridge, Massachusetts 02139.

Genes & Development
|October 1, 1991
PubMed
Summary

The MAT alpha 1 protein, not MCM1, mediates alpha-specific gene induction in yeast Saccharomyces cerevisiae in response to alpha-mating pheromone. This induction relies on specific domains within MAT alpha 1 and pheromone response pathway genes.

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Area of Science:

  • Molecular Biology
  • Yeast Genetics
  • Gene Regulation

Background:

  • In Saccharomyces cerevisiae, alpha-specific gene expression is controlled by MAT alpha 1 and MCM1.
  • These proteins bind to PQ elements upstream of alpha-specific genes.
  • PQ elements regulate both alpha-specific expression and gene induction by alpha-mating pheromone.

Purpose of the Study:

  • To investigate whether MAT alpha 1, MCM1, or both mediate PQ box-conferred induction by alpha-mating pheromone.
  • To identify the functional domains within MAT alpha 1 responsible for pheromone-induced transcriptional activation.

Main Methods:

  • Gene fusion technique using LexA DNA-binding domain.
  • Construction of MCM1-LexA and MAT alpha 1-LexA fusion proteins.
  • Analysis of trans-activation of a lacZ reporter gene.

Related Experiment Videos

  • Site-directed mutagenesis and progressive N-terminal deletions in MAT alpha 1-LexA fusions.
  • Assessment of dependency on pheromone response pathway genes (STE7, STE11, STE12).

    • Main Results:

    • MCM1-LexA fusion protein showed basal trans-activation but no induction by alpha-factor.
    • MAT alpha 1-LexA fusion protein exhibited basal trans-activation and significant induction by alpha-factor.
    • Progressive N-terminal deletions in MAT alpha 1 reduced basal activity but increased inducibility.
    • Deletion of ≥57 amino acids from MAT alpha 1's N-terminus abolished trans-activation.
    • MAT alpha 1-LexA activity was dependent on STE7, STE11, and STE12 gene functions.

    Conclusions:

    • MAT alpha 1, not MCM1, is the primary mediator of alpha-mating pheromone-induced transcriptional activation via PQ elements.
    • Specific domains within the MAT alpha 1 protein are crucial for both basal activity and pheromone inducibility.
    • The pheromone response pathway, involving STE7, STE11, and STE12, is essential for MAT alpha 1-mediated induction.