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Mol Cell Biol, April 1998, p. 2240-2251, Vol. 18, No. 4
0270-7306/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

The Tomato Hsf System: HsfA2 Needs Interaction with HsfA1 for Efficient Nuclear Import and May Be Localized in Cytoplasmic Heat Stress Granules

Klaus-Dieter Scharf, Harald Heider,dagger Ingo Höhfeld, Ruth Lyck, Enrico Schmidt, and Lutz Nover*

Department of Molecular Cell Biology, Goethe University Frankfurt, D-60439 Frankfurt/Main, Germany

Received 31 October 1997/Returned for modification 16 December 1997/Accepted 12 January 1998

In heat-stressed (HS) tomato (Lycopersicon peruvianum) cell cultures, the constitutively expressed HS transcription factor HsfA1 is complemented by two HS-inducible forms, HsfA2 and HsfB1. Because of its stability, HsfA2 accumulates to fairly high levels in the course of a prolonged HS and recovery regimen. Using immunofluorescence and cell fractionation experiments, we identified three states of HsfA2: (i) a soluble, cytoplasmic form in preinduced cultures maintained at 25°C, (ii) a salt-resistant, nuclear form found in HS cells, and (iii) a stored form of HsfA2 in cytoplasmic HS granules. The efficient nuclear transport of HsfA2 evidently requires interaction with HsfA1. When expressed in tobacco protoplasts by use of a transient-expression system, HsfA2 is mainly retained in the cytoplasm unless it is coexpressed with HsfA1. The essential parts for the interaction and nuclear cotransport of the two Hsfs are the homologous oligomerization domain (HR-A/B region of the A-type Hsfs) and functional nuclear localization signal motifs of both partners. Direct physical interaction of the two Hsfs with formation of relatively stabile hetero-oligomers was shown by a two-hybrid test in Saccharomyces cerevisiae as well as by coimmunoprecipitation using tomato and tobacco whole-cell lysates.

* Corresponding author. Mailing address: Department of Molecular Cell Biology, Biocenter N200, 30G, Goethe University Frankfurt, Marie Curie Str. 9, D-60439 Frankfurt/Main, Germany. Phone: 49- 69-798-29284. Fax: 49-69-798-29286. E-mail: nover{at}cellbiology.uni-frankfurt.de.

dagger Present address: DIBIT-Scientific Institute San Raffaele, I-20132 Milan, Italy.




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