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Cloning, Expression Characteristics and Subcellular Location of Heat Shock Transcription Factor Gene (ZmHsf06) in Zea mays |
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Abstract Heat shock transcription factors (Hsfs) widely exist in plants and play a key role under extreme environmental conditions, especially under heat shock stress. Most researches focused on model plants such as Arabidopsis and tomato (Lycopersicon esculentum). There were few reports about maize Hsfs so far. In this study, a Hsf gene, named ZmHsf06 (GenBank accession: GRMZM2G115456_T01), was cloned from maize (Zea mays) young leaves treated by heat shock at 42 ℃ for 1 h using homologous cloning methods. The patterns of ZmHsf06 expression level in different organs and its subcellular location were analyzed. Sequence analysis showed that the coding sequences (CDS) of ZmHsf06 was 1 584 bp encoding a protein of 527 amino acids. ZmHsf06 contained not only the most conserved and typical DNA-binding domain (DBD) of Hsf family, but also other functional domains such as a nuclear localization signal (NLS) KKRR peptide, a nuclear export signal (NES) IGDLTEQM peptide and an aromatic, large hydrophobic and acidic amino residues (AHA) DSFWEQFL peptide. Real-time quantitative PCR (qRT-PCR) analysis showed that under normal growth conditions, ZmHsf06 expressed in roots, stems and leaves of maize seedlings with the highest expression level in roots, and also in functional leaves, ears, immature embryo and pollen at anther period with the highest expression level in pollen and the lowest in leaves. The expression level of ZmHsf06 was up-regulated by 42 ℃ heat shock, abscisic acid(ABA) and salt stress, respectively. In details, under heat shock of 42 ℃, the highest expression level of ZmHsf06 in roots was 16 folds of that in leaves, and it appeared later than in leaves. While treated with ABA, the highest expression level in leaves was 2 folds of that in roots, and it appeared later than in roots. Under salt stress, the expression levels of ZmHsf06 in leaves and roots were increased significantly and shared the similar patterns. Based on transient expression assay using onion (Allium cepa L.) epidermis, it was found that the ZmHsf06 was located in nucleus specifically both under normal condition and heat shock at 37 ℃ within 1 h. It suggested that ZmHsf06 probably participates in signal transduction process of pollen development and responses to abiotic stresses at transcription level, and functions in nucleus. The results of this work provide basic data for isolating and characterizing more Hsfs in maize, as well as exploring their biological functions in abiotic stresses.
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Received: 21 July 2014
Published: 06 January 2015
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Agashe V R, Hartl F U. 2008. Roles of molecular chaperones in cytoplasmic protein folding[J]. Seminars In Cell & Developmental Biology, 11(1): 15-25.Aranda M A, Escaler M, Thomas C L, et al. 1999. A heat shock transcription factor in pea is differentially controlled by heat and virus replication[J]. The Plant Journal, 20(2): 153-161.Almoguera C, Rojas A, Díaz-Martín J, et al. 2002. A seed-specific heat-shock transcription factor involved in developmental regulation during embryogenesis in sunflower[J]. The Journal of Biological Chemistry, 277(46): 43866-43872.Busch W, Wunderlich M, Sch?ffl F. 2005. Identification of novel heat shock factor-dependent genes and biochemical pathways in Arabidopsis thaliana[J]. The Plant Journal, 41(1): 1-14.Brameier M, Krings A, MacCallum R M. 2007. NucPred—Predicting nuclear localization of proteins[J]. Bioinformatics Applications Note, 23(9): 1159-1160. Charng Y Y, Liu H C, Liu N Y, et al. 2007. A heat-induced transcription factor, HsfA2, is required for extension of acquired thermotolerance in Arabidopsis[J]. Plant Physiology, 143(1): 251-262.Czarnecka-Verner E, Yuan C X, For P C. 1995. Isolation and characterization of six heat shock transcription factor cDNA clones from soybean[J]. Plant Molecular Biology, 29(1): 37-51.Ellis R J. 2000. Chaperone substrates inside the cell[J]. Trends in Biochemical Science, 25: 210-212.Gagliardi D, Breton C, Chaboud A, et al. 1995. Dumas C. Expression of heat shock factor and heat shock protein 70 genes during maize pollen development[J]. Plant Molecular Biology, 29(4): 841-856.Guo J K, Wu J, Ji Q, Wang C, et al. 2008. Genome-wide analysis of heat shock transcription factor families in rice and Arabidopsis[J]. Journal of Genetics and Genomics, 35(2): 105-118.Heerklotz D, D?ring P, Bonzelius F. 2001. The balance of nuclear import and export determines the intrancellular distribution and function of tomato heat stress transcription factor HsfA2[J]. Molecular and Cellular Biology, 21(5): 1759-1768.Hübel A, Sch?ffl F. 1994. Arabidopsis heat shock factor: isolation and characterization of the gene and the recombinant protein [J]. Plant Molecular Biology, 26(1): 353-362.Jolly C, Morimoto R I. 2000. Role of the heat shock response and molecular chaperones in oncogenesis and cell death[J]. Journal of the National Cancer Institute, 92: 1564-1572.La Cour T, Kiemer L, Molgaard A, et al. 2004. Analysis and prediction of leucine-rich nuclear export signals[J]. Protein Eng Des Sel, 17: 527-536. Lee J H, Hübel A, Sch?ffl F. 1995. Derepression of the activity of genetically engineered heat shock factor causes constitutive synthesis of heat shock proteins and increased thermotolerance in transgenic Arabidopsis[J]. The Plant Journal, 8(4): 603-612.Li H C, Li G L, Liu Z H, et al. 2014. Cloning, localization and expression of ZmHsf-Like in Zea mays[J]. J Integritive Agriculture, 13(6): 1230-1238.Lin Y X, Jiang H Y, Chu Z X, et al. 2011. Genome-wide identification, classification and analysis of heat shock transcription factor family in maize[J]. BMC Genomics, 12: 76-89.Liu H C, Liao H T, Charng Y Y. 2011. The role of class A1 heat shock factors (HSFA1s) in response to heat and other stresses in Arabidopsis[J]. Plant Cell and Environment, 34(5): 738-751.Lohmann C, Eggers-Schumacher G, Wunderlich M, et al. 2004. Two different heat shock transcription factors regulate immediate early expression of stress genes in Arabidopsis [J]. Molecular Genetics and Genomics, 271(1): 11-21.Mishra S K, Tripp J, Winkelhaus S, et al. 2002. In the complex family of heat stress transcription factors, HsfA1 has a unique role as master regulator of thermotolerance in tomato[J]. Genes Development, 16(12): 1555-1567Nishizawa A, Yabuta Y, Yoshida E, et al. 2006. Arabidopsis heat shock transcription factor A2 as a key regulator in response to several types of environmental stress[J]. The Plant Journal, 48(4): 535-547Nishizawa-Yokoi A, Nosaka R, Hayashi H, et al. 2011. HsfA1d and HsfA1e involved in the transcriptional regulation of HsfA2 function as key regulators for the Hsf signaling network in response to environmental stress[J]. Plant Cell and Physiology, 52(5): 933-945.Nover L, Bharti K, D?ring P, et al. 2001. Arabidopsis and the heat stress transcription factor world: how many heat stress transcription factors do we need[J]. Cell Stress Chaperones, 6(3): 177-189.Nover L, Scharf K D, Gagliardi D, et al. 1996. The Hsf world: classification and properties of plant heat stress transcription factors[J]. Cell Stress Chaperones, 1(4): 215-223. Scharf K D, Heider H, H?hfeld I, et al. 1998. The tomato Hsf system: HsfA2 needs interaction with HsfA1 for efficient nuclear import and may be localized in cytoplasmic heat stress granules[J]. Molecular and Cellular Biology, 18(4): 2240-2251.Scharf K D, Rose S, Zott W, Sch?ffl F, Nover L. 1990. Three tomato genes code for heat stress transcription factors with a region of remarkable homology to the DNA-binding domain of the yeast HSF[J]. EMBO Journal, 9(13): 4495-4501.Sch?ffl F, Pr?ndl R, Reindl A. 1998. Regulation of the heat-shock response[J]. Plant Physiology, 117(4): 1135-1141.Schramm F, Ganguli A, Kiehlmann E, et al. 2006. The heat stress transcription factor HsfA2 serves as a regulatory amplifier of a subset of genes in the heat stress response in Arabidopsis[J]. Plant Molecular Biology, 60(5): 759-772.Shim D, Hwang J U, Lee J, et al. 2009. Orthologs of the class A4 heat shock transcription factor HsfA4a confer cadmium tolerance in wheat and rice[J]. Plant Cell, 21(12): 4031-4043.Yamanouchi U, Yano M, Lin H X, et al. 2002. A rice spotted leaf gene, Spl7, encodes a heat stress transcription factor protein [J]. Proceedings of National Academy of Science of United States of America, 99(11): 7530-7535.Yokotani N, Ichikawa T, Kondou Y, et al. 2008. Expression of rice heat stress transcription factor OsHsfA2e enhances tolerance to environmental stresses in transgenic Arabidopsis[J]. Planta, 227(5): 957-967. |
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