The Contribution of AAUAAA and the Upstream Element UUUGUA to the Efficiency of MRNA 3'-end Formation in Plants

Overview

Journal EMBO J

Specialties Biotechnology
Molecular Biology

Date 1994 May 1

PMID 8187773

Citations 46

Authors

H M Rothnie

J Reid

T Hohn

Affiliations

Soon will be listed here.

Abstract

The requirement for sequence specificity in the AAUAAA motif of the cauliflower mosaic virus (CaMV) polyadenylation signal was examined by saturation mutagenesis. While deletion of AAUAAA almost abolished processing at the CaMV polyadenylation site, none of the 18 possible single base mutations had a dramatic effect on processing efficiency. The effect of replacing all six nucleotides simultaneously varied depending on the sequence used, but some replacements were as detrimental as the deletion mutant. Taken together, these results confirm that AAUAAA is an essential component of the CaMV polyadenylation signal, but indicate that a high degree of sequence variation can be tolerated. A repeated UUUGUA motif was identified as an important upstream accessory element of the CaMV polyadenylation signal. This sequence was able to induce processing at a heterologous polyadenylation site in a sequence-specific and additive manner. The effect of altering the spacing between this upstream element and the AAUAAA was examined; moving these two elements closer together or further apart reduces the processing efficiency. The upstream element does not function to signal processing at the CaMV polyadenylation site if placed downstream of the cleavage site. Analysis of further upstream sequences revealed that almost all of the 200 nt fragment required for maximal processing contributes positively to processing efficiency. Furthermore, isolated far upstream sequences distinct from UUUGUA were also able to induce processing at a heterologous polyadenylation site.

Citing Articles

Arabidopsis and maize terminator strength is determined by GC content, polyadenylation motifs and cleavage probability.

Gorjifard S, Jores T, Tonnies J, Mueth N, Bubb K, Wrightsman T Nat Commun. 2024; 15(1):5868.

PMID: 38997252 PMC: 11245536. DOI: 10.1038/s41467-024-50174-7.

and Maize Terminator Strength is Determined by GC Content, Polyadenylation Motifs and Cleavage Probability.

Gorjifard S, Jores T, Tonnies J, Mueth N, Bubb K, Wrightsman T bioRxiv. 2023; .

PMID: 37398426 PMC: 10312805. DOI: 10.1101/2023.06.16.545379.

Plant terminators: the unsung heroes of gene expression.

de Felippes F, Waterhouse P J Exp Bot. 2022; 74(7):2239-2250.

PMID: 36477559 PMC: 10082929. DOI: 10.1093/jxb/erac467.

Transcription Terminator-Mediated Enhancement in Transgene Expression in Maize: Preponderance of the AUGAAU Motif Overlapping With Poly(A) Signals.

Wang P, Kumar S, Zeng J, McEwan R, Wright T, Gupta M Front Plant Sci. 2020; 11:570778.

PMID: 33178242 PMC: 7591816. DOI: 10.3389/fpls.2020.570778.

Plant 3' Regulatory Regions From mRNA-Encoding Genes and Their Uses to Modulate Expression.

Bernardes W, Menossi M Front Plant Sci. 2020; 11:1252.

PMID: 32922424 PMC: 7457121. DOI: 10.3389/fpls.2020.01252.

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