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. 1984 Nov;3(11):2491–2497. doi: 10.1002/j.1460-2075.1984.tb02161.x

The DNA sequence analysis of soybean heat-shock genes and identification of possible regulatory promoter elements

Fritz Schöffl 1, Eberhard Raschke 1, Ronald T Nagao 1
PMCID: PMC557717  PMID: 16453563

Abstract

The soybean possesses a gene family encoding the major low mol. wt. heat-shock proteins of 15–18 kd. We have determined the primary DNA sequences of two of the genes, both located on the same subgenomic DNA fragment. The protein coding regions are characterized by long uninterrupted open reading frames and by sequence homology of 92% and 100% with a heat-shock specific cDNA. One protein sequence deduced from the completely cloned gene hs6871 is composed of 153 amino acids with a total mol. wt. of 17.3 kd; the other protein is a truncated polypeptide containing 73 amino acids at the carboxy-terminal end of an incompletely cloned heat-shock gene designated hs6834. Investigations of the hydrophilic/hydrophobic characteristics of the polypeptides revealed a conservation of structural features between heat-shock proteins from soybean, Caenorhabditis and Drosophila and mammalian lens α-crystallin. The 5' end of the soybean heat-shock gene hs6871 was mapped by S1 nuclease at a position which is ˜100 nucleotides upstream from the translation start codon and 25 nucleotides downstream from a TATA-box sequence. Six other potential promoter elements which are homologous to the Drosophila heat-shock consensus sequence CT-GAA-TTC-AG-, are present within ˜150 nucleotides upstream from the TATA-box. The possible functions of these promoter elements in transcriptional regulation of expression of soybean heat-shock gene are discussed.

Keywords: DNA sequences, hs genes, promoter elements, soybean, S1 mapping

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Ashburner M., Bonner J. J. The induction of gene activity in drosophilia by heat shock. Cell. 1979 Jun;17(2):241–254. doi: 10.1016/0092-8674(79)90150-8. [DOI] [PubMed] [Google Scholar]
  2. Bazaral M., Helinski D. R. Circular DNA forms of colicinogenic factors E1, E2 and E3 from Escherichia coli. J Mol Biol. 1968 Sep 14;36(2):185–194. doi: 10.1016/0022-2836(68)90374-4. [DOI] [PubMed] [Google Scholar]
  3. Benoist C., O'Hare K., Breathnach R., Chambon P. The ovalbumin gene-sequence of putative control regions. Nucleic Acids Res. 1980 Jan 11;8(1):127–142. doi: 10.1093/nar/8.1.127. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Berk A. J., Sharp P. A. Sizing and mapping of early adenovirus mRNAs by gel electrophoresis of S1 endonuclease-digested hybrids. Cell. 1977 Nov;12(3):721–732. doi: 10.1016/0092-8674(77)90272-0. [DOI] [PubMed] [Google Scholar]
  5. Bienz M., Pelham H. R. Expression of a Drosophila heat-shock protein in Xenopus oocytes: conserved and divergent regulatory signals. EMBO J. 1982;1(12):1583–1588. doi: 10.1002/j.1460-2075.1982.tb01359.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Brisson N., Verma D. P. Soybean leghemoglobin gene family: normal, pseudo, and truncated genes. Proc Natl Acad Sci U S A. 1982 Jul;79(13):4055–4059. doi: 10.1073/pnas.79.13.4055. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Burke J. F., Ish-Horowicz D. Expression of Drosophila heat shock genes is regulated in Rat-cells. Nucleic Acids Res. 1982 Jul 10;10(13):3821–3830. doi: 10.1093/nar/10.13.3821. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Corces V., Holmgren R., Freund R., Morimoto R., Meselson M. Four heat shock proteins of Drosophila melanogaster coded within a 12-kilobase region in chromosome subdivision 67B. Proc Natl Acad Sci U S A. 1980 Sep;77(9):5390–5393. doi: 10.1073/pnas.77.9.5390. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Corces V., Pellicer A., Axel R., Meselson M. Integration, transcription, and control of a Drosophila heat shock gene in mouse cells. Proc Natl Acad Sci U S A. 1981 Nov;78(11):7038–7042. doi: 10.1073/pnas.78.11.7038. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Darnell J. E., Jr Variety in the level of gene control in eukaryotic cells. Nature. 1982 Jun 3;297(5865):365–371. doi: 10.1038/297365a0. [DOI] [PubMed] [Google Scholar]
  11. Dennis E. S., Gerlach W. L., Pryor A. J., Bennetzen J. L., Inglis A., Llewellyn D., Sachs M. M., Ferl R. J., Peacock W. J. Molecular analysis of the alcohol dehydrogenase (Adh1) gene of maize. Nucleic Acids Res. 1984 May 11;12(9):3983–4000. doi: 10.1093/nar/12.9.3983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Garnier J., Osguthorpe D. J., Robson B. Analysis of the accuracy and implications of simple methods for predicting the secondary structure of globular proteins. J Mol Biol. 1978 Mar 25;120(1):97–120. doi: 10.1016/0022-2836(78)90297-8. [DOI] [PubMed] [Google Scholar]
  13. Ingolia T. D., Craig E. A. Four small Drosophila heat shock proteins are related to each other and to mammalian alpha-crystallin. Proc Natl Acad Sci U S A. 1982 Apr;79(7):2360–2364. doi: 10.1073/pnas.79.7.2360. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Ingolia T. D., Craig E. A., McCarthy B. J. Sequence of three copies of the gene for the major Drosophila heat shock induced protein and their flanking regions. Cell. 1980 Oct;21(3):669–679. doi: 10.1016/0092-8674(80)90430-4. [DOI] [PubMed] [Google Scholar]
  15. Karch F., Török I., Tissières A. Extensive regions of homology in front of the two hsp70 heat shock variant genes in Drosophila melanogaster. J Mol Biol. 1981 May 25;148(3):219–230. doi: 10.1016/0022-2836(81)90536-2. [DOI] [PubMed] [Google Scholar]
  16. Key J. L., Lin C. Y., Chen Y. M. Heat shock proteins of higher plants. Proc Natl Acad Sci U S A. 1981 Jun;78(6):3526–3530. doi: 10.1073/pnas.78.6.3526. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Key J. L., Silflow C. The occurrence and distribution of poly(a) ribonucleic Acid in soybean. Plant Physiol. 1975 Sep;56(3):364–369. doi: 10.1104/pp.56.3.364. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Kozak M. Selection of initiation sites by eucaryotic ribosomes: effect of inserting AUG triplets upstream from the coding sequence for preproinsulin. Nucleic Acids Res. 1984 May 11;12(9):3873–3893. doi: 10.1093/nar/12.9.3873. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Kyte J., Doolittle R. F. A simple method for displaying the hydropathic character of a protein. J Mol Biol. 1982 May 5;157(1):105–132. doi: 10.1016/0022-2836(82)90515-0. [DOI] [PubMed] [Google Scholar]
  20. Lin C. Y., Roberts J. K., Key J. L. Acquisition of Thermotolerance in Soybean Seedlings : Synthesis and Accumulation of Heat Shock Proteins and their Cellular Localization. Plant Physiol. 1984 Jan;74(1):152–160. doi: 10.1104/pp.74.1.152. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Maxam A. M., Gilbert W. Sequencing end-labeled DNA with base-specific chemical cleavages. Methods Enzymol. 1980;65(1):499–560. doi: 10.1016/s0076-6879(80)65059-9. [DOI] [PubMed] [Google Scholar]
  22. Nagao R. T., Shah D. M., Eckenrode V. K., Meagher R. B. Multigene family of actin-related sequences isolated from a soybean genomic library. DNA. 1981;1(1):1–9. doi: 10.1089/dna.1.1981.1.1. [DOI] [PubMed] [Google Scholar]
  23. Pedersen K., Devereux J., Wilson D. R., Sheldon E., Larkins B. A. Cloning and sequence analysis reveal structural variation among related zein genes in maize. Cell. 1982 Jul;29(3):1015–1026. doi: 10.1016/0092-8674(82)90465-2. [DOI] [PubMed] [Google Scholar]
  24. Pelham H. R. A regulatory upstream promoter element in the Drosophila hsp 70 heat-shock gene. Cell. 1982 Sep;30(2):517–528. doi: 10.1016/0092-8674(82)90249-5. [DOI] [PubMed] [Google Scholar]
  25. Pelham H. R., Bienz M. A synthetic heat-shock promoter element confers heat-inducibility on the herpes simplex virus thymidine kinase gene. EMBO J. 1982;1(11):1473–1477. doi: 10.1002/j.1460-2075.1982.tb01340.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Proudfoot N. The end of the message. Nature. 1982 Aug 5;298(5874):516–517. doi: 10.1038/298516a0. [DOI] [PubMed] [Google Scholar]
  27. Pustell J., Kafatos F. C. A convenient and adaptable package of DNA sequence analysis programs for microcomputers. Nucleic Acids Res. 1982 Jan 11;10(1):51–59. doi: 10.1093/nar/10.1.51. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Russnak R. H., Jones D., Candido E. P. Cloning and analysis of cDNA sequences coding for two 16 kilodalton heat shock proteins (hsps) in Caenorhabditis elegans: homology with the small hsps of Drosophila. Nucleic Acids Res. 1983 May 25;11(10):3187–3205. doi: 10.1093/nar/11.10.3187. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Schuler M. A., Schmitt E. S., Beachy R. N. Closely related families of genes code for the alpha and alpha' subunits of the soybean 7S storage protein complex. Nucleic Acids Res. 1982 Dec 20;10(24):8225–8244. doi: 10.1093/nar/10.24.8225. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Schöffl F., Key J. L. An analysis of mRNAs for a group of heat shock proteins of soybean using cloned cDNAs. J Mol Appl Genet. 1982;1(4):301–314. [PubMed] [Google Scholar]
  31. Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
  32. Southgate R., Ayme A., Voellmy R. Nucleotide sequence analysis of the Drosophila small heat shock gene cluster at locus 67B. J Mol Biol. 1983 Mar 25;165(1):35–57. doi: 10.1016/s0022-2836(83)80241-1. [DOI] [PubMed] [Google Scholar]
  33. Van Der Ouderaa F. J., De Jong W. W., Hilderink A., Bloemendal H. The amino-acids sequence of the alphaB2 chain of bovine alpha-crystallin. Eur J Biochem. 1974 Nov 1;49(1):157–168. doi: 10.1111/j.1432-1033.1974.tb03821.x. [DOI] [PubMed] [Google Scholar]
  34. Velazquez J. M., Lindquist S. hsp70: nuclear concentration during environmental stress and cytoplasmic storage during recovery. Cell. 1984 Mar;36(3):655–662. doi: 10.1016/0092-8674(84)90345-3. [DOI] [PubMed] [Google Scholar]
  35. Vieira J., Messing J. The pUC plasmids, an M13mp7-derived system for insertion mutagenesis and sequencing with synthetic universal primers. Gene. 1982 Oct;19(3):259–268. doi: 10.1016/0378-1119(82)90015-4. [DOI] [PubMed] [Google Scholar]
  36. Vodkin L. O., Rhodes P. R., Goldberg R. B. cA lectin gene insertion has the structural features of a transposable element. Cell. 1983 Oct;34(3):1023–1031. doi: 10.1016/0092-8674(83)90560-3. [DOI] [PubMed] [Google Scholar]
  37. Voellmy R., Goldschmidt-Clermont M., Southgate R., Tissières A., Levis R., Gehring W. A DNA segment isolated from chromosomal site 67B in D. melanogaster contains four closely linked heat-shock genes. Cell. 1981 Jan;23(1):261–270. doi: 10.1016/0092-8674(81)90290-7. [DOI] [PubMed] [Google Scholar]
  38. Voellmy R., Rungger D. Transcription of a Drosophila heat shock gene is heat-induced in Xenopus oocytes. Proc Natl Acad Sci U S A. 1982 Mar;79(6):1776–1780. doi: 10.1073/pnas.79.6.1776. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Wadsworth S. C., Craig E. A., McCarthy B. J. Genes for three Drosophila heat-shock-induced proteins at a single locus. Proc Natl Acad Sci U S A. 1980 Apr;77(4):2134–2137. doi: 10.1073/pnas.77.4.2134. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Wasylyk B., Kédinger C., Corden J., Brison O., Chambon P. Specific in vitro initiation of transcription on conalbumin and ovalbumin genes and comparison with adenovirus-2 early and late genes. Nature. 1980 Jun 5;285(5764):367–373. doi: 10.1038/285367a0. [DOI] [PubMed] [Google Scholar]
  41. Wu C. Two protein-binding sites in chromatin implicated in the activation of heat-shock genes. Nature. 1984 May 17;309(5965):229–234. doi: 10.1038/309229a0. [DOI] [PubMed] [Google Scholar]
  42. van der Ouderaa F. J., de Jong W. W., Bloemendal H. The amino-acid sequence of the alphaA2 chain of bovine alpha-crystallin. Eur J Biochem. 1973 Nov 1;39(1):207–222. doi: 10.1111/j.1432-1033.1973.tb03119.x. [DOI] [PubMed] [Google Scholar]

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