N-terminomics reveals control of Arabidopsis seed storage proteins and proteases by the Arg/N-end rule pathway

A - Papers appearing in refereed journals

Zhang, H., Gannon, L., Hassall, K. L., Deery, M. J., Gibbs, D. J., Holdsworth, M. J., Van der Hoorn, R. A. L., Lilley, K. S. and Theodoulou, F. L. 2018. N-terminomics reveals control of Arabidopsis seed storage proteins and proteases by the Arg/N-end rule pathway. New Phytologist. 218 (3), pp. 1106-1126.

AuthorsZhang, H., Gannon, L., Hassall, K. L., Deery, M. J., Gibbs, D. J., Holdsworth, M. J., Van der Hoorn, R. A. L., Lilley, K. S. and Theodoulou, F. L.
Abstract

The N-end rule pathway of targeted protein degradation is an important regulator of diverse processes in plants but detailed knowledge regarding its influence on the proteome is lacking. To investigate the impact of the Arg/N-end rule pathway on the proteome of etiolated seedlings, we used terminal amine isotopic labelling of substrates with tandem mass tags (TMT-TAILS) for relative quantification of N-terminal peptides in prt6, an Arabidopsis thaliana N-end rule mutant lacking the E3 ligase PROTEOLYSIS6 (PRT6).
TMT-TAILS identified over 4000 unique N-terminal peptides representing c. 2000 protein groups. Forty-five protein groups exhibited significantly increased N-terminal peptide abundance in prt6 seedlings, including cruciferins, major seed storage proteins, which were regulated by Group VII Ethylene Response Factor (ERFVII) transcription factors, known substrates of PRT6. Mobilisation of endosperm a-cruciferin was delayed in prt6 seedlings. N-termini of several proteases were downregulated in prt6, including RD21A. RD21A transcript, protein and activity levels were downregulated in a largely ERFVII-dependent manner. By contrast, cathepsin B3 protein and activity were upregulated by ERFVIIs independent of transcript.
We propose that the PRT6 branch of the pathway regulates protease activities in a complex manner and optimises storage reserve mobilisation in the transition from seed to seedling via control of ERFVII action.

KeywordsArabidopsis thaliana; Cruciferin; N-end rule; N-terminomics; Protease; Quantitative proteomics; TAILS; Tandem mass tag (TMT)
Year of Publication2018
JournalNew Phytologist
Journal citation218 (3), pp. 1106-1126
Digital Object Identifier (DOI)doi:10.1111/nph.14909
Open accessPublished as ‘gold’ (paid) open access
FunderBiotechnology and Biological Sciences Research Council
Funder project or codeSubstrates of the N-end rule of targeted protein degradation
TPM - Tailoring Plant Metabolism - Work package 1 (WP1) - High value lipids for health and industry
Publisher's version
Output statusPublished
Publication dates
Online23 Nov 2017
Publication process dates
Accepted23 Oct 2017
PublisherWiley
Wiley-Blackwell
Copyright licenseCC BY
ISSN0028-646X

Permalink - https://repository.rothamsted.ac.uk/item/84563/n-terminomics-reveals-control-of-arabidopsis-seed-storage-proteins-and-proteases-by-the-arg-n-end-rule-pathway

23 total views
43 total downloads
0 views this month
0 downloads this month
Download files as zip