Skip to main content
NCBI home page
Plant Physiology logoLink to Plant Physiology
. 1995 Dec;109(4):1353–1361. doi: 10.1104/pp.109.4.1353

Structural requirements of oleosin domains for subcellular targeting to the oil body.

G J van Rooijen 1, M M Moloney 1
PMCID: PMC157669  PMID: 8539295

Abstract

We have investigated the protein domains responsible for the correct subcellular targeting of plant seed oleosins. We have attempted to study this targeting in vivo using "tagged" oleosins in transgenic plants. Different constructs were prepared lacking gene sequences encoding one of three structural domains of natural oleosins. Each was fused in frame to the Escherichia coli uid A gene encoding beta-glucuronidase (GUS). These constructs were introduced into Brassica napus using Agrobacterium-mediated transformation. GUS activity was measured in washed oil bodies and in the soluble protein fraction of the transgenic seeds. It was found that complete Arabidopsis oleosin-GUS fusions undergo correct subcellular targeting in transgenic Brassica seeds. Removal of the C-terminal domain of the Arabidopsis oleosin comprising the last 48 amino acids had no effect on overall subcellular targeting. In contrast, loss of the first 47 amino acids (N terminus) or amino acids 48 to 113 (which make up a lipophilic core) resulted in impaired targeting of the fusion protein to the oil bodies and greatly reduced accumulation of the fusion protein. Northern blotting revealed that this reduction is not due to differences in mRNA accumulation. Results from these measurements indicated that both the N-terminal and central oleosin domain are important for targeting to the oil body and show that there is a direct correlation between the inability to target to the oil body and protein stability.

Full Text

The Full Text of this article is available as a PDF (2.5 MB).

Selected References

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

  1. 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]
  2. Carrington J. C., Freed D. D., Leinicke A. J. Bipartite signal sequence mediates nuclear translocation of the plant potyviral NIa protein. Plant Cell. 1991 Sep;3(9):953–962. doi: 10.1105/tpc.3.9.953. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Dower W. J., Miller J. F., Ragsdale C. W. High efficiency transformation of E. coli by high voltage electroporation. Nucleic Acids Res. 1988 Jul 11;16(13):6127–6145. doi: 10.1093/nar/16.13.6127. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Iturriaga G., Jefferson R. A., Bevan M. W. Endoplasmic reticulum targeting and glycosylation of hybrid proteins in transgenic tobacco. Plant Cell. 1989 Mar;1(3):381–390. doi: 10.1105/tpc.1.3.381. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Kavanagh T. A., Jefferson R. A., Bevan M. W. Targeting a foreign protein to chloroplasts using fusions to the transit peptide of a chlorophyll a/b protein. Mol Gen Genet. 1988 Dec;215(1):38–45. doi: 10.1007/BF00331300. [DOI] [PubMed] [Google Scholar]
  6. Lee K., Bih F. Y., Learn G. H., Ting J. T., Sellers C., Huang A. H. Oleosins in the gametophytes of Pinus and Brassica and their phylogenetic relationship with those in the sporophytes of various species. Planta. 1994;193(3):461–469. doi: 10.1007/BF00201827. [DOI] [PubMed] [Google Scholar]
  7. Lee K., Ratnayake C., Huang A. H. Genetic dissection of the co-expression of genes encoding the two isoforms of oleosins in the oil bodies of maize kernel. Plant J. 1995 Apr;7(4):603–611. doi: 10.1046/j.1365-313x.1995.7040603.x. [DOI] [PubMed] [Google Scholar]
  8. Murphy D. J. Storage lipid bodies in plants and other organisms. Prog Lipid Res. 1990;29(4):299–324. [PubMed] [Google Scholar]
  9. Pang S. Z., Rasmussen J., Ye G. N., Sanford J. C. Use of the signal peptide of Pisum vicilin to translocate beta-glucuronidase in Nicotiana tabacum. Gene. 1992 Mar 15;112(2):229–234. doi: 10.1016/0378-1119(92)90381-x. [DOI] [PubMed] [Google Scholar]
  10. Plant A. L., van Rooijen G. J., Anderson C. P., Moloney M. M. Regulation of an Arabidopsis oleosin gene promoter in transgenic Brassica napus. Plant Mol Biol. 1994 May;25(2):193–205. doi: 10.1007/BF00023237. [DOI] [PubMed] [Google Scholar]
  11. Qu R., Wang S. M., Lin Y. H., Vance V. B., Huang A. H. Characteristics and biosynthesis of membrane proteins of lipid bodies in the scutella of maize (Zea mays L.). Biochem J. 1986 Apr 1;235(1):57–65. doi: 10.1042/bj2350057. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Roberts M. R., Hodge R., Ross J. H., Sorensen A., Murphy D. J., Draper J., Scott R. Characterization of a new class of oleosins suggests a male gametophyte-specific lipid storage pathway. Plant J. 1993 May;3(5):629–636. [PubMed] [Google Scholar]
  13. Vance V. B., Huang A. H. The major protein from lipid bodies of maize. Characterization and structure based on cDNA cloning. J Biol Chem. 1987 Aug 15;262(23):11275–11279. [PubMed] [Google Scholar]
  14. Verwoerd T. C., Dekker B. M., Hoekema A. A small-scale procedure for the rapid isolation of plant RNAs. Nucleic Acids Res. 1989 Mar 25;17(6):2362–2362. doi: 10.1093/nar/17.6.2362. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Yatsu L. Y., Jacks T. J. Spherosome membranes: half unit-membranes. Plant Physiol. 1972 Jun;49(6):937–943. doi: 10.1104/pp.49.6.937. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. van Rooijen G. J., Terning L. I., Moloney M. M. Nucleotide sequence of an Arabidopsis thaliana oleosin gene. Plant Mol Biol. 1992 Apr;18(6):1177–1179. doi: 10.1007/BF00047721. [DOI] [PubMed] [Google Scholar]

Articles from Plant Physiology are provided here courtesy of Oxford University Press

RESOURCES