Intracellular localization and induction of a dynamic RNA-editing event of macro-algal V-ATPase subunit A (VHA-A) in response to copper

Morris, Ceri, Owen, J. R., Thomas, M. C., El-Hiti, G. A., Harwood, John L. ORCID: https://orcid.org/0000-0003-2377-2612 and Kille, Peter ORCID: https://orcid.org/0000-0001-6023-5221 2014. Intracellular localization and induction of a dynamic RNA-editing event of macro-algal V-ATPase subunit A (VHA-A) in response to copper. Plant, Cell & Environment 37 (1) , pp. 189-203. 10.1111/pce.12145

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Abstract

A V-ATPase subunit A protein (VHA-A) transcript together with a variant (C793 to U), which introduces a stop codon truncating the subunit immediately downstream of its ATP binding site, was identified within a Fucus vesiculosus cDNA from a heavy metal contaminated site. This is intriguing because the VHA-A subunit is the crucial catalytic subunit responsible for the hydrolysis of ATP that drives ion transport underlying heavy metal detoxification pathways. We employed a chemiluminescent hybridization protection assay to quantify the proportion of both variants directly from mRNA while performing quantification of total transcript using Q-PCR. Polyclonal antisera raised against recombinant VHA-A facilitated simultaneous detection of parent and truncated VHA-A and revealed its cellular and subcellular localization. By exploiting laboratory exposures and samples from an environmental copper gradient, we showed that total VHA-A transcript and protein, together with levels of the truncated variant, were induced by copper. The absence of a genomic sequence representing the truncated variant suggests a RNA editing event causing the production of the truncated VHA-A. Based on these observations, we propose RNA editing as a novel molecular process underpinning VHA trafficking and intracellular sequestration of heavy metals under stress.

Item Type:	Article
Date Type:	Publication
Status:	Published
Schools:	Biosciences
Subjects:	Q Science > QH Natural history > QH301 Biology
Uncontrolled Keywords:	gene expression; macro-algae; Phaeophyta; protein location; toxicity
Publisher:	Wiley-Blackwell
ISSN:	0140-7791
Last Modified:	24 Oct 2022 11:58
URI:	https://orca.cardiff.ac.uk/id/eprint/49876

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