Cloning and Sequencing of the cDNA Encoding the Rubber Elongation Factor of Hevea brasiliensis

Plant Physiol. 1991 September; 97(1): 317–321.

PMCID: PMC1081000

Cloning and Sequencing of the cDNA Encoding the Rubber Elongation Factor of Hevea brasiliensis

Elisabeth Goyvaerts, Mark Dennis, David Light,¹ and Nam-Hai Chua

Institute of Molecular and Cell Biology, National University of Singapore, Singapore 0511

Department of Medicinal and Biomolecular Chemistry, Genentech, Inc., South San Francisco, California 94080

Laboratory of Plant Molecular Biology, The Rockefeller University, New York, New York 10021-6396

¹ Present address: Codon, 213 East Grand Avenue, South San Francisco, CA 94080.

This article has been cited by other articles in PMC.

Abstract

In Hevea brasiliensis, the rubber particle in the laticiferous vessel is the site of rubber (cis-1-4-polyisoprene) biosynthesis. A 14 kilodalton protein, rubber elongation factor (REF), is associated with the rubber particle in a ratio of one REF to one rubber molecule (Dennis M, Henzel W, Bell J, Kohr W, Light D [1989] J Biol Chem 264: 18618-18628; Dennis M, Light D [1989] J Biol Chem 264: 18608-18617). To obtain more information concerning the function of REF and its synthesis and assembly in the rubber particle, we isolated cDNA clones encoding REF. We used antibodies to REF to screen a Hevea leaf γgt11 cDNA expression library and obtained several positive clones. Sequence analysis of the REF cDNA clones showed that the REF mRNA contains 121 nucleotides of 5′-nontranslated sequences and a 205 nucleotide 3′-nontranslated region. The open reading frame encodes the entire 14 kilodalton REF protein without any extra amino acids (Dennis M, Henzel W, Bell J, Kohr W, Light D [1989] J Biol Chem 264: 18618-18628). The REF cDNA was subcloned in pGEM-3Z/-4Z and expressed in vitro. The translation product is a 14 kilodalton protein that can be immunoprecipitated with antibodies to REF. Addition of microsomal membranes to the in vitro translation product did not alter the mobility of the REF protein. This, and the sequence data, indicate that REF is not made as a preprotein. Our results suggest that REF is synthesized on free polysomes in the laticifer cytoplasm and that assembly of the rubber particles is likely to occur in the cytosol.

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Selected References

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