Heat stress transcription factors from tomato can functionally replace HSF1 in the yeast Saccharomyces cerevisiae.
The fact that yeast HSF1 is essential for survival under nonstress conditions can be used to test heterologous Hsfs for the ability to substitute for the endogenous protein. Our results demonstrate that like Hsf of Drosophila, tomato Hsfs A1 and A2 can functionally replace the corresponding yeast protein, but Hsf B1 cannot. In addition to survival at 28 degrees C, we checked the transformed yeast strains for temperature sensitivity of growth, induced thermotolerance and activator function using two different lacZ reporter constructs. Tests with full-length Hsfs were supplemented by assays using mutant Hsfs lacking parts of their C-terminal activator region or oligomerization domain, or containing amino acid substitutions in the DNA-binding domain. Remarkably, results with the yeast system are basically similar to those obtained by the analysis of the same Hsfs as transcriptional activators in a tobacco protoplast assay. Most surprising is the failure of HsfB1 to substitute for the yeast Hsf. The defect can be overcome by addition to HsfB1 of a short C-terminal peptide motif from HsfA2 (34 amino acid residues), which represents a type of minimal activator necessary for interaction with the yeast transcription apparatus. Deletion of the oligomerization domain (HR-A/B) does not interfere with Hsf function for survival or growth at higher temperatures. But monomeric Hsf has a markedly reduced affinity for DNA, as shown by lacZ reporter and band-shift assays.
Boscheinen
O
O
Molecular Cell Biology, Biocenter of the J.W. Goethe University, Frankfurt, Germany.
Lyck
R
R
Queitsch
C
C
Treuter
E
E
Zimarino
V
V
Scharf
K D
KD
D000595:Amino Acid Sequence; D004268:DNA-Binding Proteins / Q000235:genetics* / Q000378:metabolism; D017930:Genes, Reporter; D000076249:Heat Shock Transcription Factors; D006360:Heat-Shock Proteins / Q000235:genetics*; D018869:Heat-Shock Response / Q000235:genetics*; D006358:Hot Temperature; D018551:Solanum lycopersicum / Q000235:genetics*; D008969:Molecular Sequence Data; D009154:Mutation; D010940:Plant Proteins; D011485:Protein Binding; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D029701:Saccharomyces cerevisiae Proteins; D013045:Species Specificity; D014157:Transcription Factors / Q000235:genetics* / Q000378:metabolism
D004268:DNA-Binding Proteins; C067168:HSF1 protein, S cerevisiae; D000076249:Heat Shock Transcription Factors; D006360:Heat-Shock Proteins; D010940:Plant Proteins; D029701:Saccharomyces cerevisiae Proteins; D014157:Transcription Factors