Dataset Information

Arabidopsis, protein turnover, plastid, leaf growth and development-15N fully labelling as reference

ABSTRACT: In order to characterize plant protein degradation rates and understand their determinants as they relate to plant growth rate, we have applied 15N labelling approaches in leaves of the Arabidopsis rosette. We found a series of leaves in Arabidopsis plants for which the proteome was stable over time and degradation rates of individual proteins could thus be measured by considering the dilution of total protein abundance through growth. By progressively labelling new peptides with 15N and measuring the decrease in the abundance of over 60,000 peptides with natural isotope abundance profiles we determined the degradation rate of 1228 proteins. The exponential constant of the decay rate (KD) for each protein calculated from the relative isotope abundance of each peptide and the fold change in protein abundance during growth showed a wide distribution, ranging from 0 to 2 per day. This showed Arabidopsis protein half-lives vary from several hours to several months. In assessing intrinsic factors to explain these protein degradation rates, little effect of the N-end amino acid of proteins or of protein aggregation propensity were found, however protein complex membership and specific protein domains were strong predictors of degradation rate. We found new rapidly degrading subunits in a variety of protein complexes in plastids, identified the set of plant proteins whose degradation rate changes in different leaves of the rosette and correlated with leaf growth rate, and calculated the protein turnover energy costs in different leaves and their key determinants within the proteome.

INSTRUMENT(S): 6520A Quadrupole Time-of-Flight LC/MS

ORGANISM(S): Arabidopsis Thaliana (mouse-ear Cress)

TISSUE(S): Leaf

SUBMITTER: Lei Li

LAB HEAD: Harvey Millar

PROVIDER: PXD004549 | Pride | 2017-02-01

REPOSITORIES: Pride

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Protein Degradation Rate in Arabidopsis thaliana Leaf Growth and Development.

Li Lei L Nelson Clark J CJ Trösch Josua J Castleden Ian I Huang Shaobai S Millar A Harvey AH

The Plant cell 20170130 2

We applied 15N labeling approaches to leaves of the Arabidopsis thaliana rosette to characterize their protein degradation rate and understand its determinants. The progressive labeling of new peptides with 15N and measuring the decrease in the abundance of >60,000 existing peptides over time allowed us to define the degradation rate of 1228 proteins in vivo. We show that Arabidopsis protein half-lives vary from several hours to several months based on the exponentia ...[more]

PMID: 28138016

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Project description:Bacterial adaptation involves extensive cellular reorganization. In particular, growth rate adjustments are associated with substantial modifications of gene expression and mRNA abundance. In this work we aimed to assess the role of mRNA degradation during such variations. A genome-wide transcriptomic-based method was used to determine mRNA half-lives. The model bacterium Lactococcus lactis was used and five growth rates were studied in continuous cultures under isoleucine-limitation and in batch cultures during the adaptation to the isoleucine starvation. During continuous isoleucine-limited growth, the mRNAs of different genes had different half-lives. The stability of most of the transcripts was not constant, and increased as the growth rate decreased. This half-life diversity was analyzed to investigate determinants of mRNA stability. The concentration, length, codon adaptation index and secondary structures of mRNAs were found to contribute to the regulation of mRNA stability in these conditions. However, the growth rate was, by far, the most influential determinant. The respective influences of mRNA degradation and transcription on the regulation of intra-cellular transcript concentration were estimated. The role of degradation on mRNA homeostasis was clearly evidenced: for more than 90 % of the mRNAs studied during continuous isoleucine-limited growth of L. lactis, degradation was antagonistic to transcription. Although both transcription and degradation had, opposite effects,, the mRNA changes in response to growth rate were driven by transcription. Interestingly, degradation control increased during the dynamic adaptation of bacteria as the growth rate reduced due to progressive isoleucine starvation in batch cultures. This work shows that mRNA decay differs between gene transcripts and according to the growth rate. It demonstrates that mRNA degradation is an important regulatory process involved in bacterial adaptation. However, its impact on the regulation of mRNA levels is smaller than that of transcription in the conditions studied. In the study presented here mRNA stabilities were analyzed at 5 growth rates. For each growth rate mRNA levels were measured in a time course experiment following rifampicin addition. At least 12 time points per growth rate are available, including 3 replicates of the zero.

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	21DL3R1F1.mzML	Mzml
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Dataset Information

Arabidopsis, protein turnover, plastid, leaf growth and development-15N fully labelling as reference

Dataset's files

Publications

Protein Degradation Rate in <i>Arabidopsis thaliana</i> Leaf Growth and Development.

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