Abstract
Heat shock transcription factor (HSF) is a multidomain protein that exists as a monomer under normal conditions and is reversibly induced upon heat shock to a trimeric state that binds to DNA with high affinity. The maintenance of the monomeric state is dependent on hydrophobic heptad repeats located at the amino- and carboxy-terminal regions which have been proposed to form an intramolecular coiled-coil structure. In a systematic deletion analysis to identify other regions of HSF that may be required to regulate its oligomeric state, we have found that local sequences encompassing the carboxy-terminal end of the DNA binding domain and a broad region of HSF between the heptad repeats also contribute to this regulation. Immunocytochemical analysis of mutant HSF proteins revealed a canonical motif required for nuclear localization. HSF proteins lacking the nuclear localization signal remain in the cytoplasm, but these HSFs nonetheless exhibit reversible heat stress-inducible trimerization. The results indicate that the signals that regulate HSF trimerization operate in both the nuclear and cytoplasmic compartments of the cell.
