Publications by authors named "Michael Holdsworth"

Reduced oxygen availability (hypoxia) represents a key plant abiotic stress in natural and agricultural systems, but conversely it is also an important component of normal growth and development. We review recent advances that demonstrate how genetic adaptations associated with hypoxia impact the known plant oxygen-sensing mechanism through the PLANT CYSTEINE OXIDASE N-degron pathway. Only 3 protein substrates of this pathway have been identified, and all adaptations identified to date are associated with the most important of these, the group VII ETHYLENE RESPONSE FACTOR transcription factors.

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Oxygen is a key signalling component of plant biology, and whilst an oxygen-sensing mechanism was previously described in Arabidopsis thaliana, key features of the associated PLANT CYSTEINE OXIDASE (PCO) N-degron pathway and Group VII ETHYLENE RESPONSE FACTOR (ERFVII) transcription factor substrates remain untested or unknown. We demonstrate that ERFVIIs show non-autonomous activation of root hypoxia tolerance and are essential for root development and survival under oxygen limiting conditions in soil. We determine the combined effects of ERFVIIs in controlling gene expression and define genetic and environmental components required for proteasome-dependent oxygen-regulated stability of ERFVIIs through the PCO N-degron pathway.

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Increasing drought phenomena pose a serious threat to agricultural productivity. Although plants have multiple ways to respond to the complexity of drought stress, the underlying mechanisms of stress sensing and signaling remain unclear. The role of the vasculature, in particular the phloem, in facilitating inter-organ communication is critical and poorly understood.

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Flowering plants (angiosperms) can grow at extreme altitudes, and have been observed growing as high as 6,400 metres above sea level; however, the molecular mechanisms that enable plant adaptation specifically to altitude are unknown. One distinguishing feature of increasing altitude is a reduction in the partial pressure of oxygen (pO). Here we investigated the relationship between altitude and oxygen sensing in relation to chlorophyll biosynthesis-which requires molecular oxygen-and hypoxia-related gene expression.

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Article Synopsis
  • - The research reveals that genetic changes in specific regulatory elements (WT box and W box) of the RAP2.12 gene link to how certain species adapt to varying environmental stresses like drought and flooding.
  • - Different transcription factors control these regulatory elements, leading to the selective accumulation of gene transcripts that respond to low oxygen levels.
  • - The study suggests that the transition between different gene regulatory variants is tied to species moving from dry to more humid environments, highlighting a complex adaptation strategy influenced by noncoding genetic variations.
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  • * Overexpression of UCP1 in Arabidopsis thaliana leads to changes in nuclear gene expression and affects physiological responses by inhibiting the cytoplasmic PLANT CYSTEINE OXIDASE (PCO) branch of the PROTEOLYSIS (PRT)6 N-degron pathway, which is essential for sensing oxygen levels.
  • * This research uncovers a new mechanism where mitochondrial signaling represses the PCO N-de
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The N-degron pathway is a branch of the ubiquitin-proteasome system where amino-terminal residues serve as degradation signals. In a synthetic biology approach, we expressed ubiquitin ligase PRT6 and ubiquitin conjugating enzyme 2 (AtUBC2) from in a strain with mutation in its endogenous N-degron pathway. The two enzymes re-constitute part of the plant N-degron pathway and were probed by monitoring the stability of co-expressed GFP-linked plant proteins starting with Arginine N-degrons.

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Article Synopsis
  • Aerobic respiration is crucial for survival in eukaryotes, and how organisms sense oxygen plays a key role in their ability to thrive.
  • Plants and animals have developed different mechanisms for oxygen sensing, including the destruction of transcription factors and direct oxygen detection by specific enzymes.
  • Understanding these sensing systems can yield insights for developing therapeutic strategies for humans and creating crops that can better withstand stress.
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  • VERNALIZATION2 (VRN2) is a key protein in plant development that is regulated by oxygen and is part of the polycomb repressive complex 2 (PRC2), impacting how plants respond to environmental factors.
  • In Arabidopsis thaliana, VRN2 affects flowering time based on light conditions and shapes root architecture by negatively influencing root growth.
  • Its activity is enhanced in cold conditions, allowing it to function effectively outside of growth regions, where it interacts with other proteins for proper developmental processes.
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N-degron pathways of ubiquitin-mediated proteolysis (formerly known as the N-end rule pathway) control the stability of substrate proteins dependent on the amino-terminal (Nt) residue. Unlike yeast or mammalian N-recognin E3 ligases, which each recognize several different classes of Nt residues, in , N-recognin functions of different N-degron pathways are carried out independently by PROTEOLYSIS (PRT)1, PRT6, and other unknown proteins. PRT1 recognizes type 2 aromatic Nt-destabilizing residues and PRT6 recognizes type 1 basic residues.

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Each year, billions of songbirds cross large ecological barriers during their migration. Understanding how they perform this incredible task is crucial to predict how global change may threaten the safety of such journeys. Earlier studies based on radar suggested that most songbirds cross deserts in intermittent flights at high altitude, stopping in the desert during the day, while recent tracking with light loggers suggested diurnal prolongation of nocturnal flights and common non-stop flights for some species.

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Article Synopsis
  • - Masson et al. (2019) explored how responses to low oxygen (hypoxia) are influenced by the degradation of specific proteins in different kingdoms like animals and plants.
  • - They discovered a mammalian version of an oxygen-sensing pathway that had only been seen in plants before, highlighting a cross-kingdom similarity.
  • - This research, along with other recent studies, offers fresh perspectives on how eukaryotic organisms detect and respond to oxygen levels.
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The amino-terminal residue of a protein (or amino-terminus of a peptide following protease cleavage) can be an important determinant of its stability, through the Ubiquitin Proteasome System associated N-degron pathways. Plants contain a unique combination of N-degron pathways (previously called the N-end rule pathways) E3 ligases, PROTEOLYSIS (PRT)6 and PRT1, recognizing non-overlapping sets of amino-terminal residues, and others remain to be identified. Although only very few substrates of PRT1 or PRT6 have been identified, substrates of the oxygen and nitric oxide sensing branch of the PRT6 N-degron pathway include key nuclear-located transcription factors (ETHYLENE RESPONSE FACTOR VIIs and LITTLE ZIPPER 2) and the histone-modifying Polycomb Repressive Complex 2 component VERNALIZATION 2.

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Article Synopsis
  • Timely detection of environmental changes is crucial for plant survival, particularly during submergence when gas levels fluctuate. !* -
  • In the study, Arabidopsis thaliana plants sense submergence through ethylene trapping, which enhances the stability of ERFVII transcription factors amid flooding-induced low oxygen levels. !* -
  • The findings indicate that ethylene can increase PHYTOGLOBIN1 levels to deplete nitric oxide, thereby helping plants pre-adapt to low oxygen conditions, revealing important targets for creating flood-resistant crops. !*
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Assessing posttranslational modification (PTM) patterns within protein molecules and reading their functional implications present grand challenges for plant biology. We combine four perspectives on PTMs and their roles by considering five classes of PTMs as examples of the broader context of PTMs. These include modifications of the N terminus, glycosylation, phosphorylation, oxidation, and N-terminal and protein modifiers linked to protein degradation.

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Article Synopsis
  • The study explores how the polycomb repressive complex 2 (PRC2) regulates gene repression in flowering plants and identifies a new mechanism for its control related to oxygen levels.
  • Researchers discovered that the plant PRC2 subunit VRN2 has an N-terminal degron that triggers its degradation, shaped by early angiosperm evolution through gene duplication.
  • The findings suggest that environmental factors like hypoxia and cold exposure increase VRN2 levels by interfering with its degradation, potentially linking environmental signals to the epigenetic regulation of plant growth and development.
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A key component of seed germination is the interplay of mechanical forces governing embryo growth and the surrounding restraining endosperm tissue. Endosperm cell separation is therefore thought to play a critical role in the control of this developmental transition. Here we demonstrate that in Arabidopsis thaliana seeds, endosperm cell expansion is a key component of germination.

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Article Synopsis
  • The Arg/N-end rule pathway influences plant development, especially in transforming dormant seeds into photosynthesizing seedlings, by regulating protein degradation.
  • PRT6, an N-recognin E3 ligase, degrades certain transcription factors (ERFVIIs) that control sugar sensitivity and oil body breakdown in seedlings.
  • The study suggests that the Arg/N-end rule pathway operates through both ABA-dependent and independent mechanisms, highlighting its role in multiple processes during the seed-to-seedling transition.
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The N-end rule pathway is a highly conserved constituent of the ubiquitin proteasome system, yet little is known about its biological roles. Here we explored the role of the N-end rule pathway in the plant immune response. We investigated the genetic influences of components of the pathway and known protein substrates on physiological, biochemical and metabolic responses to pathogen infection.

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Post-translational modifications are essential mediators between stimuli from development or the environment and adaptive transcriptional patterns. Recent data allow a first glimpse at how two modifications, phosphorylation and sumoylation, act interdependently to modulate stress responses. In particular, many components of the SUMO conjugation system are phosphoproteins, and some regulators and enzymes of protein phosphorylation can be sumoylated.

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N-term 2017 was the first international meeting to bring together researchers from diverse disciplines with a shared interest in protein N-terminal modifications and the N-end rule pathway of ubiquitin-mediated proteolysis, providing a platform for interdisciplinary cross-kingdom discussions and collaborations, as well as strengthening the visibility of this growing scientific community.

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