Home LiteratureArticle Details
PMID: 11238913 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

The balance of nuclear import and export determines the intracellular distribution and function of tomato heat stress transcription factor HsfA2.

Molecular and cellular biology ·Vol. 21 ·No. 5 ·2001-03-00 ·Pages 1759-68

Heerklotz D, Döring P, Bonzelius F, Winkelhaus S, Nover L

Abstract

Tomato heat stress transcription factor HsfA2 is a shuttling protein with dominant cytoplasmic localization as a result of a nuclear import combined with an efficient export. Besides the nuclear localization signal (NLS) adjacent to the oligomerization domain, a C-terminal leucine-rich motif functions as a nuclear export signal (NES). Mutant forms of HsfA2 with a defective or an absent NES are nuclear proteins. The same is true for the wild-type HsfA2 if coexpressed with HsfA1 or in the presence of export inhibitor leptomycin B (LMB). Fusion of the NES domain of HsfA2 to HsfB1, which is a nuclear protein, caused export of the HsfB1-A2NES hybrid protein, and this effect was reversed by the addition of LMB. Due to the lack of background problems, Chinese hamster ovary (CHO) cells represent an excellent system for expression and functional analysis of tomato Hsfs. The results faithfully reflect the situation found in plant cells (tobacco protoplasts). The intriguing role of NLS and NES accessibility for the intracellular distribution of HsfA2 is underlined by the results of heat stress treatments of CHO cells (41 degrees C). Despite the fact that nuclear import and export are not markedly affected, HsfA2 remains completely cytoplasmic at 41 degrees C even in the presence of LMB. The temperature-dependent conformational transition of HsfA2 with shielding of the NLS evidently needs intramolecular interaction between the internal HR-A/B and the C-terminal HR-C regions. It is not observed with the HR oligomerization domain (HR-A/B region) deletion form of HsfA2 or in HsfA2-HsfA1 hetero-oligomers.

MeSH Terms
Animals Antibiotics, Antineoplastic/pharmacology Blotting, Western CHO Cells Cell Nucleus/metabolism Cricetinae Cytoplasm/metabolism DNA-Binding Proteins/chemistry,metabolism Fatty Acids, Unsaturated/pharmacology Fluorescent Antibody Technique, Indirect Heat Shock Transcription Factors Heat-Shock Proteins Luciferases/metabolism Lycopersicon esculentum/metabolism Microscopy, Fluorescence Models, Genetic Plant Proteins Plasmids/metabolism Protein Binding Protein Conformation Protein Structure, Tertiary Protein Transport Temperature Time Factors Transcription Factors/chemistry,metabolism Transfection Transformation, Genetic
Chemicals
Antibiotics, Antineoplastic DNA-Binding Proteins Fatty Acids, Unsaturated Heat Shock Transcription Factors Heat-Shock Proteins Plant Proteins Transcription Factors Luciferases leptomycin B
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Heerklotz D
Department of Molecular Cell Biology, Biocenter, Goethe-University Frankfurt, Frankfurt am Main, Germany.
Döring P
Bonzelius F
Winkelhaus S
Nover L
References (47)
47 references, click to expand
  1. Maintenance of G2 arrest in the Xenopus oocyte: a role for 14-3-3-mediated inhibition of Cdc25 nuclear import.
    EMBO J. 1999 Apr 15;18(8):2174-83 PMID: 10205171
  2. Conservation of a stress response: human heat shock transcription factors functionally substitute for yeast HSF.
    EMBO J. 1997 Nov 3;16(21):6466-77 PMID: 9351828
  3. Enhanced green fluorescence by the expression of an Aequorea victoria green fluorescent protein mutant in mono- and dicotyledonous plant cells.
    Proc Natl Acad Sci U S A. 1996 Jun 11;93(12):5888-93 PMID: 8650188
  4. The HIV-1 Rev protein.
    Annu Rev Microbiol. 1998;52:491-532 PMID: 9891806
  5. Intracellular distribution and identification of the nuclear localization signals of two plant heat-stress transcription factors.
    Planta. 1997;202(1):117-25 PMID: 9177056
  6. Nucleocytoplasmic transport: the last 200 nanometers.
    Cell. 1998 Feb 6;92(3):327-36 PMID: 9476893
  7. RanGTP-regulated interactions of CRM1 with nucleoporins and a shuttling DEAD-box helicase.
    Mol Cell Biol. 1999 Sep;19(9):6276-85 PMID: 10454574
  8. Leptomycin B inactivates CRM1/exportin 1 by covalent modification at a cysteine residue in the central conserved region.
    Proc Natl Acad Sci U S A. 1999 Aug 3;96(16):9112-7 PMID: 10430904
  9. Nuclear export of proteins and RNAs.
    Curr Opin Cell Biol. 1997 Jun;9(3):420-9 PMID: 9159083
  10. Transport between the cell nucleus and the cytoplasm.
    Annu Rev Cell Dev Biol. 1999;15:607-60 PMID: 10611974
  11. Arabidopsis heat shock factor is constitutively active in Drosophila and human cells.
    Mol Gen Genet. 1995 Jul 28;248(2):136-41 PMID: 7651336
  12. Nuclear export of proteins in plants: AtXPO1 is the export receptor for leucine-rich nuclear export signals in Arabidopsis thaliana.
    Plant J. 1999 Dec;20(6):695-705 PMID: 10652141
  13. Regulation of the heat shock transcriptional response: cross talk between a family of heat shock factors, molecular chaperones, and negative regulators.
    Genes Dev. 1998 Dec 15;12(24):3788-96 PMID: 9869631
  14. NF-AT activation requires suppression of Crm1-dependent export by calcineurin.
    Nature. 1999 Mar 18;398(6724):256-60 PMID: 10094050
  15. The Hsf world: classification and properties of plant heat stress transcription factors.
    Cell Stress Chaperones. 1996 Dec;1(4):215-23 PMID: 9222607
  16. Exportin 1 (Crm1p) is an essential nuclear export factor.
    Cell. 1997 Sep 19;90(6):1041-50 PMID: 9323132
  17. Isolation and characterization of HsfA3, a new heat stress transcription factor of Lycopersicon peruvianum.
    Plant J. 2000 May;22(4):355-65 PMID: 10849352
  18. Structural analysis of 14-3-3 phosphopeptide complexes identifies a dual role for the nuclear export signal of 14-3-3 in ligand binding.
    Mol Cell. 1999 Aug;4(2):153-66 PMID: 10488331
  19. Binding of 14-3-3 proteins and nuclear export control the intracellular localization of the mitotic inducer Cdc25.
    Genes Dev. 1999 May 1;13(9):1067-72 PMID: 10323858
  20. Oxidative stress abolishes leptomycin B-sensitive nuclear export of transcription repressor Bach2 that counteracts activation of Maf recognition element.
    J Biol Chem. 2000 May 19;275(20):15370-6 PMID: 10809773
  21. Molecular cloning and expression of a hexameric Drosophila heat shock factor subject to negative regulation.
    Cell. 1990 Nov 30;63(5):1085-97 PMID: 2257625
  22. Transient expression and heat-stress-induced co-aggregation of endogenous and heterologous small heat-stress proteins in tobacco protoplasts.
    Plant J. 2000 Nov;24(3):397-411 PMID: 11069712
  23. Expression of human heat shock transcription factors 1 and 2 in HeLa cells and yeast.
    Cell Stress Chaperones. 1997 Dec;2(4):263-75 PMID: 9495283
  24. Nuclear import in permeabilized protoplasts from higher plants has unique features.
    Plant Cell. 1996 Aug;8(8):1337-52 PMID: 8776900
  25. Heat stress proteins and transcription factors.
    Cell Mol Life Sci. 1997 Jan;53(1):80-103 PMID: 9118000
  26. Cytoplasmic heat shock granules are formed from precursor particles and are associated with a specific set of mRNAs.
    Mol Cell Biol. 1989 Mar;9(3):1298-308 PMID: 2725500
  27. Activation of the DNA-binding ability of human heat shock transcription factor 1 may involve the transition from an intramolecular to an intermolecular triple-stranded coiled-coil structure.
    Mol Cell Biol. 1994 Nov;14(11):7557-68 PMID: 7935471
  28. Three tomato genes code for heat stress transcription factors with a region of remarkable homology to the DNA-binding domain of the yeast HSF.
    EMBO J. 1990 Dec;9(13):4495-501 PMID: 2148291
  29. The role of AHA motifs in the activator function of tomato heat stress transcription factors HsfA1 and HsfA2.
    Plant Cell. 2000 Feb;12(2):265-78 PMID: 10662862
  30. Regulation of RelA subcellular localization by a putative nuclear export signal and p50.
    Mol Cell Biol. 1999 Oct;19(10):7088-95 PMID: 10490645
  31. A leucine-rich nuclear export signal in the p53 tetramerization domain: regulation of subcellular localization and p53 activity by NES masking.
    EMBO J. 1999 Mar 15;18(6):1660-72 PMID: 10075936
  32. Heat stress transcription factors from tomato can functionally replace HSF1 in the yeast Saccharomyces cerevisiae.
    Mol Gen Genet. 1997 Jul;255(3):322-31 PMID: 9268023
  33. Nuclear export in plants. Use of geminivirus movement proteins for a cell-based export assay.
    Plant Cell. 1999 Jul;11(7):1267-76 PMID: 10402428
  34. Plants contain a novel multi-member class of heat shock factors without transcriptional activator potential.
    Plant Mol Biol. 2000 Jul;43(4):459-71 PMID: 11052198
  35. Heat shock transcription factors: structure and regulation.
    Annu Rev Cell Dev Biol. 1995;11:441-69 PMID: 8689565
  36. Signal-mediated nuclear export pathways of proteins and RNAs.
    Trends Cell Biol. 1996 Aug;6(8):290-3 PMID: 15157436
  37. Promoter specificity and deletion analysis of three heat stress transcription factors of tomato.
    Mol Gen Genet. 1993 Jul;240(1):113-25 PMID: 8341257
  38. Nuclear retention of IkappaBalpha protects it from signal-induced degradation and inhibits nuclear factor kappaB transcriptional activation.
    J Biol Chem. 1999 Mar 26;274(13):9108-15 PMID: 10085161
  39. E1a transactivation of the human HSP70 promoter is mediated through the basal transcriptional complex.
    Mol Cell Biol. 1989 Jun;9(6):2574-87 PMID: 2474756
  40. Nuclear export of NF-ATc enhanced by glycogen synthase kinase-3.
    Science. 1997 Mar 28;275(5308):1930-4 PMID: 9072970
  41. Cytoplasmic sequestration of rel proteins by IkappaBalpha requires CRM1-dependent nuclear export.
    Mol Cell Biol. 2000 Mar;20(6):2269-84 PMID: 10688673
  42. CRM1 is an export receptor for leucine-rich nuclear export signals.
    Cell. 1997 Sep 19;90(6):1051-60 PMID: 9323133
  43. The tomato Hsf system: HsfA2 needs interaction with HsfA1 for efficient nuclear import and may be localized in cytoplasmic heat stress granules.
    Mol Cell Biol. 1998 Apr;18(4):2240-51 PMID: 9528795
  44. Leptomycin B inhibition of signal-mediated nuclear export by direct binding to CRM1.
    Exp Cell Res. 1998 Aug 1;242(2):540-7 PMID: 9683540
  45. Versatile cloning vectors for transient gene expression and direct gene transfer in plant cells.
    Nucleic Acids Res. 1988 Sep 12;16(17):8725 PMID: 2843828
  46. The function of plant heat shock promoter elements in the regulated expression of chimaeric genes in transgenic tobacco.
    Mol Gen Genet. 1989 Jun;217(2-3):246-53 PMID: 2770695
  47. Heat shock factor is required for growth at normal temperatures in the fission yeast Schizosaccharomyces pombe.
    Mol Cell Biol. 1993 Feb;13(2):749-61 PMID: 8423799
Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/MCB.21.5.1759-1768.2001
Published
2001-03-00
Pages
1759-68
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC86729
Subset
IM
External Links
PubMed source DOI: 10.1128/MCB.21.5.1759-1768.2001 PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: [email protected]

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2