The maize heat shock factor-binding protein paralogs EMP2 and HSBP2 interact non-redundantly with specific heat shock factors

• DOI: 10.1007/s00425-005-0191-y
• Publication Date: 2005-12-05T14:46:42Z
• Authors: Suneng Fu; Peter Rogowsky; Lutz Nover; Michael J. Scanlon
• Keywords: 2. Zero hunger; 0301 basic medicine; Binding Sites; 330; Arabidopsis Proteins; Amino Acid Motifs; Molecular Sequence Data; [SDV.BDLR]Life Sciences [q-bio]/Reproductive Biology; Zea mays; 004; Protein Structure, Tertiary; DNA-Binding Proteins; 03 medical and health sciences; Heat Shock Transcription Factors; Two-Hybrid System Techniques; Mutagenesis, Site-Directed; Protein Isoforms; Amino Acid Sequence; Carrier Proteins; Sequence Alignment; Heat-Shock Proteins; Phylogeny; Plant Proteins; Transcription Factors

The heat shock response (HSR) is a conserved mechanism by which transcripts of heat shock protein (hsp) genes accumulate following mobilization of heat shock transcription factors (HSFs) in response to thermal stress. Studies in animals identified the heat shock factor-binding protein1 (HSBP1) that interacts with heat shock transcription factor1 (HSF1) during heat shock attenuation; overexpression analyses revealed that the coiled-coil protein HSBP1 functions as a negative regulator of the HSR. Zea mays contains two HSBP paralogs, EMP2 and HSBP2, which exhibit differential accumulation during the HSR and plant development. Embryo-lethal recessive emp2 mutations revealed that EMP2 is required for the down-regulation of hsp transcription during embryogenesis, whereas accumulation of HSBP2 is induced in seedlings following heat shock. Notwithstanding, no interaction has yet been demonstrated between a plant HSBP and a plant HSF. In this report 22 maize HSF isoforms are identified comprising three structural classes: HSF-A, HSF-B and HSF-C. Phylogenetic analysis of Arabidopsis, maize and rice HSFs reveals that at least nine ancestral HSF isoforms were present prior to the separation of monocot and eudicots, followed by differential amplification of HSF members in these lineages. Yeast two-hybrid analyses show that EMP2 and HSBP2 interact non-redundantly with specific HSF-A isoforms. Site-specific mutagenesis of HSBP2 reveals that interactions between hydrophobic residues within the coiled coil are required for HSF::HSBP2 binding; domain swapping demonstrate that the isoform specificity of HSF::HSBP interaction is conferred by residues outside of the coiled coil. These data suggest that the non-redundant functions of the maize HSBPs may be explained, at least in part, by the specificity of HSBP::HSF interactions during plant development.