Arabidopsis heat shock factor: isolation and characterization of the gene and the recombinant protein.
We have isolated the Arabidopsis heat shock factor gene Athsf1 as genomic and corresponding cDNA sequences via cross-hybridization with tomato clones. Sequence analysis indicates only a partial homology with the HSFs from tomato and other organisms which is confined to the DNA-binding and the oligomerization domains. The gene is constitutively expressed but the level of mRNA for Athsf1 increases two-fold upon heat shock. However, the putative promoter region lacks the canonical heat shock elements. After expression in Escherichia coli the recombinant Athsf1 protein binds specifically to a synthetic oligonucleotide containing five heat shock elements. The native size of recombinant ATHSF1 in vitro is consistent with a trimer as demonstrated by chemical cross-linking and pore exclusion limit analysis.
Hübel
A
A
Universität Tübingen, Biologisches Institut, Germany.
Schöffl
F
F
D000595:Amino Acid Sequence; D017360:Arabidopsis / Q000235:genetics*; D029681:Arabidopsis Proteins; D001483:Base Sequence; D003001:Cloning, Molecular; D004268:DNA-Binding Proteins; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D017343:Genes, Plant / Q000235:genetics*; D000076249:Heat Shock Transcription Factors; D006360:Heat-Shock Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D006358:Hot Temperature; D018551:Solanum lycopersicum / Q000235:genetics; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010940:Plant Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D011131:Polyribonucleotides / Q000138:chemical synthesis / Q000378:metabolism; D011487:Protein Conformation; D012333:RNA, Messenger / Q000032:analysis; D018749:RNA, Plant / Q000032:analysis; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism*; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D014157:Transcription Factors
D029681:Arabidopsis Proteins; D004268:DNA-Binding Proteins; D000076249:Heat Shock Transcription Factors; D006360:Heat-Shock Proteins; D010940:Plant Proteins; D011131:Polyribonucleotides; D012333:RNA, Messenger; D018749:RNA, Plant; D011994:Recombinant Proteins; D014157:Transcription Factors