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Title: Tubulin C-terminal Post-translational Modifications Do Not Occur in Wood Forming Tissue of Populus

Abstract

Cortical microtubules (MTs) are evolutionarily conserved cytoskeletal components with specialized roles in plants, including regulation of cell wall biogenesis. MT functions and dynamics are dictated by the composition of their monomeric subunits, α- (TUA) and β-tubulins (TUB), which in animals and protists are subject to both transcriptional regulation and post-translational modifications (PTM). While spatiotemporal regulation of tubulin gene expression has been reported in plants, whether and to what extent tubulin PTMs occur in these species remain poorly understood. We chose the woody perennial Populus for investigation of tubulin PTMs in this study, with a particular focus on developing xylem where high tubulin transcript levels support MT-dependent secondary cell wall deposition. Mass spectrometry and immunodetection concurred that detyrosination, non-tyrosination and glutamylation were essentially absent in tubulins isolated from wood-forming tissues of P. deltoides and P. tremula ×alba. Label-free quantification of tubulin isotypes and RNA-Seq estimation of tubulin transcript abundance were largely consistent with transcriptional regulation. However, two TUB isotypes were detected at noticeably lower levels than expected based on RNA-Seq transcript abundance in both Populus species. These findings led us to conclude that MT composition during wood formation depends exclusively on transcriptional and, to a lesser extent, translational regulation of tubulinmore » isotypes.« less

Authors:
 [1];  [1];  [1];  [1];  [1];  [1]
  1. Univ. of Georgia, Athens, GA (United States)
Publication Date:
Research Org.:
Univ. of Georgia, Athens, GA (United States)
Sponsoring Org.:
USDOE Office of Science (SC), Biological and Environmental Research (BER)
OSTI Identifier:
1423767
Grant/Contract Number:  
SC0008470
Resource Type:
Accepted Manuscript
Journal Name:
Frontiers in Plant Science
Additional Journal Information:
Journal Volume: 7; Journal ID: ISSN 1664-462X
Publisher:
Frontiers Research Foundation
Country of Publication:
United States
Language:
English
Subject:
54 ENVIRONMENTAL SCIENCES

Citation Formats

Hu, Hao, Gu, Xi, Xue, Liang-Jiao, Swamy, Prashant S., Harding, Scott A., and Tsai, Chung-Jui. Tubulin C-terminal Post-translational Modifications Do Not Occur in Wood Forming Tissue of Populus. United States: N. p., 2016. Web. doi:10.3389/fpls.2016.01493.
Hu, Hao, Gu, Xi, Xue, Liang-Jiao, Swamy, Prashant S., Harding, Scott A., & Tsai, Chung-Jui. Tubulin C-terminal Post-translational Modifications Do Not Occur in Wood Forming Tissue of Populus. United States. https://doi.org/10.3389/fpls.2016.01493
Hu, Hao, Gu, Xi, Xue, Liang-Jiao, Swamy, Prashant S., Harding, Scott A., and Tsai, Chung-Jui. Thu . "Tubulin C-terminal Post-translational Modifications Do Not Occur in Wood Forming Tissue of Populus". United States. https://doi.org/10.3389/fpls.2016.01493. https://www.osti.gov/servlets/purl/1423767.
@article{osti_1423767,
title = {Tubulin C-terminal Post-translational Modifications Do Not Occur in Wood Forming Tissue of Populus},
author = {Hu, Hao and Gu, Xi and Xue, Liang-Jiao and Swamy, Prashant S. and Harding, Scott A. and Tsai, Chung-Jui},
abstractNote = {Cortical microtubules (MTs) are evolutionarily conserved cytoskeletal components with specialized roles in plants, including regulation of cell wall biogenesis. MT functions and dynamics are dictated by the composition of their monomeric subunits, α- (TUA) and β-tubulins (TUB), which in animals and protists are subject to both transcriptional regulation and post-translational modifications (PTM). While spatiotemporal regulation of tubulin gene expression has been reported in plants, whether and to what extent tubulin PTMs occur in these species remain poorly understood. We chose the woody perennial Populus for investigation of tubulin PTMs in this study, with a particular focus on developing xylem where high tubulin transcript levels support MT-dependent secondary cell wall deposition. Mass spectrometry and immunodetection concurred that detyrosination, non-tyrosination and glutamylation were essentially absent in tubulins isolated from wood-forming tissues of P. deltoides and P. tremula ×alba. Label-free quantification of tubulin isotypes and RNA-Seq estimation of tubulin transcript abundance were largely consistent with transcriptional regulation. However, two TUB isotypes were detected at noticeably lower levels than expected based on RNA-Seq transcript abundance in both Populus species. These findings led us to conclude that MT composition during wood formation depends exclusively on transcriptional and, to a lesser extent, translational regulation of tubulin isotypes.},
doi = {10.3389/fpls.2016.01493},
journal = {Frontiers in Plant Science},
number = ,
volume = 7,
place = {United States},
year = {2016},
month = {10}
}

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Cited by: 3 works
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Figures / Tables:

FIGURE 1 FIGURE 1: Tubulin purification. (A) SDS-PAGE analysis of xylem protein extracts from various steps of the tubulin purification process. M, markers; CP, crude proteins; FT, flow-through; W, washed, unbound proteins; EP, eluted proteins enriched with tubulins. (B) A two-color Western blot of purified tubulins from P. deltoides (Pd) and P.more » tremula x alba (Pta) normal wood (NW) or tension wood (TW) xylem probed with anti-TUA (red channel) or anti-TUB (green channel) antibodies. The scanning intensities were adjusted in the upper panel (reduced for red and increased for green) in order to visualize both TUA and TUB signals clearly. The lower panel was scanned at the same intensity for both channels.« less

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Works referencing / citing this record:

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Figures/Tables have been extracted from DOE-funded journal article accepted manuscripts.