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. 1993 Jul;5(7):769–780. doi: 10.1105/tpc.5.7.769

Cellular Concentrations and Uniformity of Cell-Type Accumulation of Two Lea Proteins in Cotton Embryos.

JK Roberts 1, NA DeSimone 1, WL Lingle 1, L Dure 3rd 1
PMCID: PMC160315  PMID: 12271086

Abstract

The levels and cell-type distribution of late embryogenesis abundant (Lea) proteins D-7 and D-113 have been determined in mature cotton embryos by immunochemical methods. The two proteins were expressed in and purified from Escherichia coli and utilized for antibody production in rabbits. The antiserum to each protein was found to interact with all members of each protein family in cotton extracts by protein gel blotting. Using these antibodies in quantitative "rocket" immunoelectrophoreses, D-7 proteins were found to accumulate to ~8 x 1015 molecules per embryo, which is equivalent to ~109 molecules per "average cell." D-113 proteins accumulate to ~1016 molecules per embryo, which equates to ~1.3 x 109 molecules per average cell. These values calculate to concentrations of about 226 and 283 [mu]M, respectively, in the cell aqueous phase immediately prior to seed desiccation. In immunocytochemical studies using the fluorophor rhodamine linked to the secondary antibody, both proteins appeared to be evenly present in the cytosol of all cell types present in the embryo, including both cotyledon and axis epidermal cells. Thus, their function does not appear related to unique functions of specific cell or tissue types. The very high molar concentrations of the two proteins, coupled with their unusual predicted structure and their cytosol location, would seem to reduce the number of their conceivable functions.

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Selected References

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  1. Almoguera C., Jordano J. Developmental and environmental concurrent expression of sunflower dry-seed-stored low-molecular-weight heat-shock protein and Lea mRNAs. Plant Mol Biol. 1992 Aug;19(5):781–792. doi: 10.1007/BF00027074. [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. Cohen A., Bray E. A. Nucleotide sequence of an ABA-induced tomato gene that is expressed in wilted vegetative organs and developing seeds. Plant Mol Biol. 1992 Jan;18(2):411–413. doi: 10.1007/BF00034969. [DOI] [PubMed] [Google Scholar]
  4. Curry J., Morris C. F., Walker-Simmons M. K. Sequence analysis of a cDNA encoding a group 3 LEA mRNA inducible by ABA or dehydration stress in wheat. Plant Mol Biol. 1991 Jun;16(6):1073–1076. doi: 10.1007/BF00016078. [DOI] [PubMed] [Google Scholar]
  5. Dure L., 3rd A repeating 11-mer amino acid motif and plant desiccation. Plant J. 1993 Mar;3(3):363–369. doi: 10.1046/j.1365-313x.1993.t01-19-00999.x. [DOI] [PubMed] [Google Scholar]
  6. Dure L., 3rd, Greenway S. C., Galau G. A. Developmental biochemistry of cottonseed embryogenesis and germination: changing messenger ribonucleic acid populations as shown by in vitro and in vivo protein synthesis. Biochemistry. 1981 Jul 7;20(14):4162–4168. doi: 10.1021/bi00517a033. [DOI] [PubMed] [Google Scholar]
  7. 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]
  8. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  9. Laurell C. B. Quantitative estimation of proteins by electrophoresis in agarose gel containing antibodies. Anal Biochem. 1966 Apr;15(1):45–52. doi: 10.1016/0003-2697(66)90246-6. [DOI] [PubMed] [Google Scholar]
  10. Marcotte W. R., Jr, Russell S. H., Quatrano R. S. Abscisic acid-responsive sequences from the em gene of wheat. Plant Cell. 1989 Oct;1(10):969–976. doi: 10.1105/tpc.1.10.969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Mundy J., Yamaguchi-Shinozaki K., Chua N. H. Nuclear proteins bind conserved elements in the abscisic acid-responsive promoter of a rice rab gene. Proc Natl Acad Sci U S A. 1990 Feb;87(4):1406–1410. doi: 10.1073/pnas.87.4.1406. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. 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]
  13. Rosenberg A. H., Lade B. N., Chui D. S., Lin S. W., Dunn J. J., Studier F. W. Vectors for selective expression of cloned DNAs by T7 RNA polymerase. Gene. 1987;56(1):125–135. doi: 10.1016/0378-1119(87)90165-x. [DOI] [PubMed] [Google Scholar]
  14. Seffens W. S., Almoguera C., Wilde H. D., Vonder Haar R. A., Thomas T. L. Molecular analysis of a phylogenetically conserved carrot gene: developmental and environmental regulation. Dev Genet. 1990;11(1):65–76. doi: 10.1002/dvg.1020110108. [DOI] [PubMed] [Google Scholar]
  15. Skriver K., Mundy J. Gene expression in response to abscisic acid and osmotic stress. Plant Cell. 1990 Jun;2(6):503–512. doi: 10.1105/tpc.2.6.503. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Walbot V., Dure L. S., 3rd Developmental biochemistry of cotton seed embryogenesis and germination. VII. Characterization of the cotton genome. J Mol Biol. 1976 Mar 15;101(4):503–536. doi: 10.1016/0022-2836(76)90242-4. [DOI] [PubMed] [Google Scholar]

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