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. 1983;2(3):419–426. doi: 10.1002/j.1460-2075.1983.tb01439.x

Identification of sequences involved in the polyadenylation of higher plant nuclear transcripts using Agrobacterium T-DNA genes as models.

P Dhaese 1, H De Greve 1, J Gielen 1, L Seurinck 1, M Van Montagu 1, J Schell 1
PMCID: PMC555149  PMID: 11894958

Abstract

Sequences in the 3'-untranslated region of two different octopine T-DNA genes were analyzed with regard to their significance in polyadenylation. Poly(A) addition sites were localized precisely by S1 nuclease mapping with T-DNA-derived mRNAs isolated from tobacco. The gene encoding transcript 7' contains two AATAAA hexanucleotides, respectively 119 bp and 170 bp downstream of the TAA stop codon. A single poly(A) site was mapped 24-25 bp downstream of the first AATAAA. Further, we show that a mutant octopine synthase gene, which has lost part of its 3'-untranslated region by deletion, is still active. This mutant gene terminates 19 bp upstream from the major wild-type polyadenylation site. The deletion also removes the AATAAT signal preceding this site. The mutant octopine synthase gene contains a minimum of four different poly(A) sites. The most prominent of these sites is identical to the minor poly(A) site of the wild-type gene, and is preceded by a sequence AATGAATATA. Three other sites are located within the adjacent plant DNA, giving rise to hybrid T-DNA/plant DNA transcripts. The two most distal sites are probably dependent on a motif AATAAATAAA, found 29 bp away from the T-DNA/plant DNA junction.

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Selected References

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