Abstract
In plants, small heat shock proteins (sHsps) have been localized to the cytosol, mitochondria, chloroplasts, and endoplasmic reticulum. UsingNtHSP24.6, a cDNA clone for tobacco mitochondrial sHsp, as a probe, we performed genomic DNA blot analysis and identified presence of a small gene family for mitochondrial sHsp in tobacco (Nicotiana tabacum). Several putative genomic clones for mitochondrial sHsp were isolated when a tobacco genomic library was screened. After restriction mapping of seven genomic clones, three that had shown different maps were selected, and their nucleotide sequences for the putative coding sequence were determined. From these three independent clones, one identical nucleotide sequence was obtained that had two exons with one intron. RNA blot hybridization of heat-stressed tobacco plants revealed a typical heat-shock-responsive accumulation ofNtHSP24.6 transcript. Under severe heat-shock conditions, an additional band was apparent, but of a larger transcript size. When we compared the amino acid sequence of NtHSP24.6 with mitochondrial sHsps from various other species, we found a high level of homology throughout the ORF, with an almost complete match to the carboxyl terminus. Our comparison of NtHSP24.6 with tobacco cytosolic class I sHsps also resulted in high homology between their α-crystalline domains, but significant divergence in their amino-terminus regions.
Similar content being viewed by others
Literature Cited
Banzet N, Richaud C, Deveaux Y, Kazmaier M, Gagnon J, Triantaphylides C. (1998) Accumulation of small heat shock proteins, including mitochondrial HSP22, induced by oxidative stress and adaptive response in tomato cells. Plant J 13: 519–527
Beissinger M, Buchner J (1998) How chaperones fold proteins. Biol Chem379: 245–259
Carranco R, Almoguera C, Jordano J (1997) A plant small heat shock protein gene expressed during zygotic embryogenesis but noninducible by heat stress. J Biol Chem272: 27470–27475
Caspers GJ, Leunissen JA, de Jong WW (1995) The expanding small heat-shock protein family and structure predictions of the conserved “alpha-crystallin domain”. J Mol Evol40: 238–248
Chen Q, Vierling E (1991) Analysis of conserved domains identifies a unique structural feature of chloroplast heat shock proteins. Mol Gen Genet226: 425–431
Choi G, Hong CB (2000) Analyses of expressed sequence tags of anther and anther-specific cDNA clones inNicotiana tabacum. J Plant Biol43: 107–113
Chou M, Chen YM, Lin YM, Lin CY (1989) Thermotolerance of isolated mitochondria associated with heat shock proteins. Plant Physiol98: 617–621
Claros MG, Vincens P (1996) Computational method to predict mitochondrially imported proteins and their targeting sequences. Eur J Biochem241: 779–786
de Jong WW, Caspers GJ, Leunissen JAM (1998) Genealogy of the α-crystallin small heat-shock protein superfamily. Intl J Biol Macromol22: 151–162
Downs CA, Heckathorn SA (1998) The mitochondrial small heat-shock protein protects NADH:ubiquinone oxidoreductase of the electron transport chain during heat stress in plants. FEBS Lett430: 246–250
Harndahl U, Hall RB, Osteryoung, KW, Vierling E, Born-man JF, Sundby C (1999) The chloroplast small heat shock protein undergoes oxidation-dependent conformational changes and may protect plants from oxidative stress. Cell Stress Chaperones4: 129–138
Hsieh MH, Chen JT, Jin TL, Chen YM, Lin CY (1992) A class of soybean low molecular weight heat shock protein. Plant Physiol99: 1279–1284.
Jakob U, Muse W, Eser M, Bardwell JCA (1999) Chaperone activity with a redox switch. Cell96: 341–352
Kyte J, Doolittle RF (1982) A simple method for displaying the hydropathic character of a protein. J Mol Biol157: 105–132
Lenne C, Block MA, Garin J, Douce R (1995) Sequence and expression of the mRNA encoding HSP22, the mitochondrial small heat-shock protein in pea leaves. Biochem J311: 805–813
Liu J, Shono M (1999) Characterization of mitochondrialocated small heat shock protein from tomato(Lycopersicon esculentum). Plant Cell Physiol40: 1297–1304
Narberhaus F (2002) α-Crystallin-type heat shock proteins: Socializing minichaperones in the context of a multichaperone network. MMBR66: 64–93
Osteryoung KW, Sundberg H, Vierling E (1993) Poly(A) tail length of a heat-shock protein RNA is increased by severe heat-stress, but intron splicing is unaffected. Mol Gen Genet239: 323–333
Park SM, Hong CB (1998) Comparison of the structure and expression pattern for a low molecular weight heatshock protein cDNA clone fromNicotiana tabacum. Mol Cells8: 594–599
Richmond CS, Glasner JD, Mau R, Jin H, Blattner FR (1999) Genome-wide expression profiling inEscherichia coli K-12. Nucleic Acids Res27: 3821–3835
Sabehat A, Weiss D, Lurie S (1996) The correlation between heat-shock protein accumulation and persistence and chilling tolerance in tomato fruit. Plant Physiol110:531–537
Sambrook J, Fritsch EF, Maniatis T (1989) Molecular Cloning: A Laboratory Manual. 2nd ed, Cold Spring Harbor Laboratory, New York
Scharf KD, Siddique M, Vierling E (2001) The expanding family ofArabidopsis thaliana small heat stress proteins and a new family of proteins containing α-crystallin domains (Acd proteins). Cell Stress Chaperones6: 225–237
Shono M, Liu J, Sanmiya K, Singh I, Din J, Suzuki K, Tsuk- aguchi T, Egawa Y (2002) Functional analysis of mitochondrial small heat shock protein. JIRCAS Working Rep 17–23
Spiess C, Beil A, Ehrmann M (1999) A temperature-dependent switch from chaperone to protease in a widely conserved heat shock protein. Cell97: 339–347
Ukaji N, Kuwabara C, Takezawa D, Arakawa K, Yoshida S, Fujikawa S (1999) Accumulation of small heat shock protein homologs in the endoplasmic reticulum of cortical parenchyma cells in mulberry in association with seasonal cold acclimation. Plant Physiol120: 481–490
van Montfort R, Slingsby C, Vierling E (2002) Structure and function of the small heat shock protein/α-crystallin family of molecular chaperones. Adv Prot Chem9: 105–156
Visioli G, Maestri E, Marmiroli N (1997) Differential display-mediated isolation of a genomic sequence for a putative mitochondrial LMW HSP specifically expressed in condition of induced thermotolerance inArabidopsis thaliana (L) Heynh. Plant Mol Biol34: 517–527
Willett DA, Basha E, Vierling E (1996) Nucleotide sequence of a cDNA encoding a mitochondrion-localized small Hsp fromArabidopsis thaliana: AtHsp23.6 (Accession No. U72958)(PCR96-117). Plant Physiol 112: 1399
Youn KM (2002) Structure and expression pattern of mitochondrial small heat-shock protein gene inNicotiana tabacum. M.S. thesis. Seoul National University, Seoul
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Koo, H.J., Xia, X. & Hong, C.B. Genes and expression pattern of tobacco mitochondrial small heat shock protein under high-temperature stress. J. Plant Biol. 46, 204–210 (2003). https://doi.org/10.1007/BF03030450
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF03030450