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. 1989 Dec;1(12):1137–1146. doi: 10.1105/tpc.1.12.1137

Novel regulation of heat shock genes during carrot somatic embryo development.

J L Zimmerman 1, N Apuya 1, K Darwish 1, C O'Carroll 1
PMCID: PMC159849  PMID: 2535535

Abstract

We have determined that somatic embryos of carrot exhibit a number of interesting and unusual properties when exposed to heat shock at different times in their development. Specifically, we have seen that mid-globular embryos can be arrested irreversibly in their development when heat-shocked, whereas all other stages of embryogenesis, both before and after this stage, are fully capable of normal development after the stress. In investigating the molecular basis of this developmental sensitivity to heat shock, using a cloned heat shock gene encoding a small heat shock protein, we have determined that globular embryos both synthesize and accumulate significantly less heat shock mRNA when compared with embryos of any other stage or to callus suspension cells. In fact, there appears to be no transcriptional induction of heat shock gene expression in response to heat shock during this time period; the gene is expressed at the same relatively low level both before and after heat shock. However, in spite of the low level of heat shock mRNA available, globular embryos synthesize the full complement of heat shock proteins in response to heat treatment. The globular embryos appear to accomplish this by translating the existing heat shock mRNAs at an elevated rate, which compensates for the low level of available mRNA. Once the embryos have progressed beyond the globular stage of development, regulation at the transcriptional level resumes, and the embryos again exhibit normal development after heat shock.

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Selected References

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  1. Bond U., Schlesinger M. J. Heat-shock proteins and development. Adv Genet. 1987;24:1–29. doi: 10.1016/s0065-2660(08)60005-x. [DOI] [PubMed] [Google Scholar]
  2. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1016/0003-2697(76)90527-3. [DOI] [PubMed] [Google Scholar]
  3. Elsdale T., Pearson M., Whitehead M. Abnormalities in somite segmentation following heat shock to Xenopus embryos. J Embryol Exp Morphol. 1976 Jun;35(3):625–635. [PubMed] [Google Scholar]
  4. Feinberg A. P., Vogelstein B. "A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity". Addendum. Anal Biochem. 1984 Feb;137(1):266–267. doi: 10.1016/0003-2697(84)90381-6. [DOI] [PubMed] [Google Scholar]
  5. Graziosi G., de Cristini F., di Marcotullio A., Marzari R., Micali F., Savoini A. Morphological and molecular modifications induced by heat shock in Drosophila melanogaster embryos. J Embryol Exp Morphol. 1983 Oct;77:167–182. [PubMed] [Google Scholar]
  6. Gurley W. B., Czarnecka E., Nagao R. T., Key J. L. Upstream sequences required for efficient expression of a soybean heat shock gene. Mol Cell Biol. 1986 Feb;6(2):559–565. doi: 10.1128/mcb.6.2.559. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Heikkila J. J., Kloc M., Bury J., Schultz G. A., Browder L. W. Acquisition of the heat-shock response and thermotolerance during early development of Xenopus laevis. Dev Biol. 1985 Feb;107(2):483–489. doi: 10.1016/0012-1606(85)90329-x. [DOI] [PubMed] [Google Scholar]
  8. Hwang C. H., Zimmerman J. L. The Heat Shock Response of Carrot : Protein Variations between Cultured Cell Lines. Plant Physiol. 1989 Oct;91(2):552–558. doi: 10.1104/pp.91.2.552. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Lindquist S. Regulation of protein synthesis during heat shock. Nature. 1981 Sep 24;293(5830):311–314. doi: 10.1038/293311a0. [DOI] [PubMed] [Google Scholar]
  10. Lindquist S. The heat-shock response. Annu Rev Biochem. 1986;55:1151–1191. doi: 10.1146/annurev.bi.55.070186.005443. [DOI] [PubMed] [Google Scholar]
  11. Luthe D. S., Quatrano R. S. Transcription in Isolated Wheat Nuclei: I. ISOLATION OF NUCLEI AND ELIMINATION OF ENDOGENOUS RIBONUCLEASE ACTIVITY. Plant Physiol. 1980 Feb;65(2):305–308. doi: 10.1104/pp.65.2.305. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. McGarry T. J., Lindquist S. The preferential translation of Drosophila hsp70 mRNA requires sequences in the untranslated leader. Cell. 1985 Oct;42(3):903–911. doi: 10.1016/0092-8674(85)90286-7. [DOI] [PubMed] [Google Scholar]
  13. Mitchell H. K., Lipps L. S. Heat shock and phenocopy induction in Drosophila. Cell. 1978 Nov;15(3):907–918. doi: 10.1016/0092-8674(78)90275-1. [DOI] [PubMed] [Google Scholar]
  14. O'Farrell P. H. High resolution two-dimensional electrophoresis of proteins. J Biol Chem. 1975 May 25;250(10):4007–4021. [PMC free article] [PubMed] [Google Scholar]
  15. Parker C. S., Topol J. A Drosophila RNA polymerase II transcription factor binds to the regulatory site of an hsp 70 gene. Cell. 1984 May;37(1):273–283. doi: 10.1016/0092-8674(84)90323-4. [DOI] [PubMed] [Google Scholar]
  16. Percival-Smith A., Segall J. Isolation of DNA sequences preferentially expressed during sporulation in Saccharomyces cerevisiae. Mol Cell Biol. 1984 Jan;4(1):142–150. doi: 10.1128/mcb.4.1.142. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Petersen N. S., Mitchell H. K. The induction of a multiple wing hair phenocopy by heat shock in mutant heterozygotes. Dev Biol. 1987 Jun;121(2):335–341. doi: 10.1016/0012-1606(87)90169-2. [DOI] [PubMed] [Google Scholar]
  18. Petko L., Lindquist S. Hsp26 is not required for growth at high temperatures, nor for thermotolerance, spore development, or germination. Cell. 1986 Jun 20;45(6):885–894. doi: 10.1016/0092-8674(86)90563-5. [DOI] [PubMed] [Google Scholar]
  19. Roccheri M. C., Sconzo G., Di Carlo M., Di Bernardo M. G., Pirrone A., Gambino R., Giudice G. Heat-shock proteins in sea urchin embryos. Transcriptional and posttranscriptional regulation. Differentiation. 1982;22(3):175–178. doi: 10.1111/j.1432-0436.1982.tb01246.x. [DOI] [PubMed] [Google Scholar]
  20. Santamaría P. Heat shock induced phenocopies of dominant mutants of the bithorax complex in Drosophila melanogaster. Mol Gen Genet. 1979 May 4;172(2):161–163. doi: 10.1007/BF00268277. [DOI] [PubMed] [Google Scholar]
  21. Sirotkin K., Davidson N. Developmentally regulated transcription from Drosophila melanogaster chromosomal site 67B. Dev Biol. 1982 Jan;89(1):196–210. doi: 10.1016/0012-1606(82)90307-4. [DOI] [PubMed] [Google Scholar]
  22. Sollner-Webb B., Reeder R. H. The nucleotide sequence of the initiation and termination sites for ribosomal RNA transcription in X. laevis. Cell. 1979 Oct;18(2):485–499. doi: 10.1016/0092-8674(79)90066-7. [DOI] [PubMed] [Google Scholar]
  23. Storti R. V., Scott M. P., Rich A., Pardue M. L. Translational control of protein synthesis in response to heat shock in D. melanogaster cells. Cell. 1980 Dec;22(3):825–834. doi: 10.1016/0092-8674(80)90559-0. [DOI] [PubMed] [Google Scholar]
  24. Widholm J. M. The use of fluorescein diacetate and phenosafranine for determining viability of cultured plant cells. Stain Technol. 1972 Jul;47(4):189–194. doi: 10.3109/10520297209116483. [DOI] [PubMed] [Google Scholar]
  25. Wittig S., Hensse S., Keitel C., Elsner C., Wittig B. Heat shock gene expression is regulated during teratocarcinoma cell differentiation and early embryonic development. Dev Biol. 1983 Apr;96(2):507–514. doi: 10.1016/0012-1606(83)90187-2. [DOI] [PubMed] [Google Scholar]
  26. Wu C. An exonuclease protection assay reveals heat-shock element and TATA box DNA-binding proteins in crude nuclear extracts. Nature. 1985 Sep 5;317(6032):84–87. doi: 10.1038/317084a0. [DOI] [PubMed] [Google Scholar]

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