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Saccharomyces cerevisiae HSP70 heat shock elements are functionally distinct

M R Young1, E A Craig

  • 1McArdle Laboratory for Cancer Research, University of Wisconsin, Madison 53706.

Molecular and Cellular Biology
|September 1, 1993
PubMed
Summary

Two heat shock elements (HSEs) in the Saccharomyces cerevisiae SSA1 gene, HSE2 and HSE3, drive transcription. HSE2 is active under basal conditions, while both HSE2 and HSE3 are crucial for heat-induced SSA1 gene expression.

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Area of Science:

  • Molecular Biology
  • Yeast Genetics
  • Gene Regulation

Background:

  • The Saccharomyces cerevisiae HSP70 gene SSA1 is regulated by multiple heat shock elements (HSEs).
  • Understanding the specific roles of individual HSEs is crucial for elucidating gene expression control.

Purpose of the Study:

  • To investigate the functional significance of each HSE in the SSA1 gene promoter.
  • To determine how HSEs contribute to both basal and heat-induced transcription.

Main Methods:

  • Site-directed mutagenesis of HSEs within the SSA1 promoter.
  • Analysis of gene expression using reporter constructs.
  • Gel mobility shift assays to assess transcription factor binding.

Main Results:

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  • Two HSE-like sequences (HSE2 and HSE3) were identified as active promoter elements.
  • HSE2 was active under basal growth conditions, while both HSE2 and HSE3 drove transcription after heat shock.
  • Basal transcription driven by HSE3 was subject to repression by flanking sequences, unlike HSE2.

Conclusions:

  • HSE2 and HSE3 play distinct but cooperative roles in regulating SSA1 expression.
  • The ability to drive basal transcription is not a universal feature of all HSEs.
  • Heat shock transcription factor binding does not solely determine the basal activity of HSEs.