Home LiteratureArticle Details
PMID: 16667403 Published · ppublish English Journal Article

Expression and Processing of an Arabidopsis 2S Albumin in Transgenic Tobacco.

Plant physiology ·Vol. 92 ·No. 4 ·1990-04-00 ·Pages 899-907

De Clercq A, Vandewiele M, De Rycke R, Van Damme J, Van Montagu M, Krebbers E, Vandekerckhove J

Abstract

2S albumin seed storage proteins undergo a complex series of posttranslational proteolytic cleavages. In order to determine if this process is correctly carried out in transgenic plants, the gene AT2S1 encoding an Arabidopsis thaliana 2S albumin isoform has been expressed in transgenic tobacco. Initial experiments using a reporter gene demonstrated that the AT2S1 promoter directs seed specific expression in both transgenic tobacco and Brassica napus plants. The entire AT2S1 gene was then transferred into tobacco plants, where it showed a tissue specific and developmentally regulated expression. Arabidopsis 2S albumin accumulates up to 0.1% of the total high-salt extractable seed protein. Protein sequencing demonstrated that the amino termini of the two Arabidopsis 2S albumin subunits were correctly processed, suggesting that the protease(s) necessary for posttranslational processing of 2S albumin precursors may display common specificities among different dicot plant species. Immunocytochemical studies showed that the Arabidopsis 2S albumin is localized in the protein body matrix of tobacco endosperm and embryo. Correct processing and targeting of the 2S albumin in transgenic plants suggests that modified versions could be expressed, allowing the study of 2S albumin processing and in particular the possible roles of the processed fragments in protein stability and/or targeting.

Authors & Affiliations
7 authors, click to expand affiliations / ORCID
De Clercq A
Laboratorium voor Genetica, Rijksuniversiteit Gent, B-9000 Gent, Belgium.
Vandewiele M
De Rycke R
Van Damme J
Van Montagu M
Krebbers E
Vandekerckhove J
References (28)
28 references, click to expand
  1. Accumulation and assembly of soybean beta-conglycinin in seeds of transformed petunia plants.
    EMBO J. 1985 Dec 1;4(12):3047-53 PMID: 16453646
  2. A quantitative dot-immunobinding assay for proteins using nitrocellulose membrane filters.
    Proc Natl Acad Sci U S A. 1984 Mar;81(6):1684-7 PMID: 6424121
  3. A standardized vector system for manipulation and enhanced expression of genes in Escherichia coli.
    DNA. 1987 Dec;6(6):583-91 PMID: 3322744
  4. Developmentally regulated expression of the bean beta-phaseolin gene in tobacco seed.
    Proc Natl Acad Sci U S A. 1985 May;82(10):3320-4 PMID: 16578787
  5. Synthesis and protein body deposition of maize 15-kd zein in transgenic tobacco seeds.
    EMBO J. 1987 Nov;6(11):3213-21 PMID: 16453803
  6. Engineering herbicide resistance in plants by expression of a detoxifying enzyme.
    EMBO J. 1987 Sep;6(9):2513-8 PMID: 16453789
  7. Correct targeting of the bean storage protein phaseolin in the seeds of transformed tobacco.
    Plant Physiol. 1985 Sep;79(1):65-71 PMID: 16664403
  8. Transformation of Brassica napus and Brassica oleracea Using Agrobacterium tumefaciens and the Expression of the bar and neo Genes in the Transgenic Plants.
    Plant Physiol. 1989 Oct;91(2):694-701 PMID: 16667089
  9. The synthesis of a 19 kilodalton zein protein in transgenic petunia plants.
    Plant Physiol. 1988 Dec;88(4):1002-7 PMID: 16666411
  10. The amino-acid sequence of the 2S sulphur-rich proteins from seeds of Brazil nut (Bertholletia excelsa H.B.K.).
    Eur J Biochem. 1986 Sep 15;159(3):597-604 PMID: 3758080
  11. Cleavage of structural proteins during the assembly of the head of bacteriophage T4.
    Nature. 1970 Aug 15;227(5259):680-5 PMID: 5432063
  12. A simple and general method for transferring genes into plants.
    Science. 1985 Mar 8;227(4691):1229-31 PMID: 17757866
  13. Nuclear transcriptional activity of the tobacco plastid psbA promoter.
    Nucleic Acids Res. 1989 Jan 11;17(1):19-29 PMID: 2563150
  14. High resolution two-dimensional electrophoresis of basic as well as acidic proteins.
    Cell. 1977 Dec;12(4):1133-41 PMID: 23215
  15. cDNA clones for Brassica napus seed storage proteins: evidence from nucleotide sequence analysis that both subunits of napin are cleaved from a precursor polypeptide.
    J Mol Appl Genet. 1983;2(3):273-83 PMID: 6689334
  16. Nucleotide sequence of a member of the napin storage protein family from Brassica napus.
    J Biol Chem. 1987 Sep 5;262(25):12202-8 PMID: 3040733
  17. Characterization of the herbicide-resistance gene bar from Streptomyces hygroscopicus.
    EMBO J. 1987 Sep;6(9):2519-23 PMID: 16453790
  18. Soybean seed lectin gene and flanking nonseed protein genes are developmentally regulated in transformed tobacco plants.
    Proc Natl Acad Sci U S A. 1986 Nov;83(21):8240-4 PMID: 3464951
  19. Efficient in vitro synthesis of biologically active RNA and RNA hybridization probes from plasmids containing a bacteriophage SP6 promoter.
    Nucleic Acids Res. 1984 Sep 25;12(18):7035-56 PMID: 6091052
  20. Regulation of gene expression during plant embryogenesis.
    Cell. 1989 Jan 27;56(2):149-60 PMID: 2643470
  21. Determination of the Processing Sites of an Arabidopsis 2S Albumin and Characterization of the Complete Gene Family.
    Plant Physiol. 1988 Aug;87(4):859-66 PMID: 16666238
  22. Efficient oligonucleotide-directed construction of mutations in expression vectors by the gapped duplex DNA method using alternating selectable markers.
    Nucleic Acids Res. 1989 Jun 26;17(12):4441-54 PMID: 2501754
  23. Cellular localization of soybean storage protein mRNA in transformed tobacco seeds.
    Proc Natl Acad Sci U S A. 1988 Jan;85(2):458-62 PMID: 16593906
  24. Properties, biosynthesis and processing of a sulfur-rich protein in Brazil nut (Bertholletia excelsa H.B.K.).
    Eur J Biochem. 1987 Feb 2;162(3):477-83 PMID: 3830150
  25. Albumin storage proteins in the protein bodies of castor bean.
    Plant Physiol. 1978 Jan;61(1):13-6 PMID: 16660226
  26. Structure of the rapeseed 1.7 S storage protein, napin, and its precursor.
    J Biol Chem. 1986 Nov 5;261(31):14576-81 PMID: 3771543
  27. Alterations in the phenotype of plant cells studied by NH(2)-terminal amino acid-sequence analysis of proteins electroblotted from two-dimensional gel-separated total extracts.
    Proc Natl Acad Sci U S A. 1987 Jul;84(14):4806-10 PMID: 16578810
  28. Expression of a maize storage protein gene in petunia plants is not restricted to seeds.
    Plant Physiol. 1988 Apr;86(4):1281-5 PMID: 16666067
Article Info
Journal
Plant physiology
Abbr.
Plant Physiol
ISSN
0032-0889
Published
1990-04-00
Pages
899-907
Language
English
Region
United States
NLM ID
0401224
PMCID
PMC1062393
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: [email protected]