PubMed Health⌕ Search

Biomedical subjects

Peter McCourt

Publications and source records attributed to Peter McCourt.

16 recordsLinked to original sources
PubMed2006

Plant hormone receptors: perception is everything.

Despite the bewildering ability of higher plants to change their development with respect to the environment, there appear to be only a few hormones that function to organize growth and development. With the recent identification of three plant hormone receptors, the molecular identities of all the major plant receptors are now known. Some plant hormones such as cytokinins, ethylene, and brassinosteroids (BR) use well-characterized signaling modules such as those involving receptor kinases, but in the case of the ethylene and BR receptors, there appear to be additional functions aside from the hormone they perceive. Auxin and gibberellin perception require unique mechanisms where the receptors are components involved in ubiquitination-dependent proteolysis. With plant hormone receptors in hand, comparisons can now be made between plants and other kingdoms as to how hormones control growth and development.

Models, Biological↗
PubMed2006

A small-molecule screen in C. elegans yields a new calcium channel antagonist.

Small-molecule inhibitors of protein function are powerful tools for biological analysis and can lead to the development of new drugs. However, a major bottleneck in generating useful small-molecule tools is target identification. Here we show that Caenorhabditis elegans can provide a platform for both the discovery of new bioactive compounds and target identification. We screened 14,100 small molecules for bioactivity in wild-type worms and identified 308 compounds that induce a variety of phenotypes. One compound that we named nemadipine-A induces marked defects in morphology and egg-laying. Nemadipine-A resembles a class of widely prescribed anti-hypertension drugs called the 1,4-dihydropyridines (DHPs) that antagonize the alpha1-subunit of L-type calcium channels. Through a genetic suppressor screen, we identified egl-19 as the sole candidate target of nemadipine-A, a conclusion that is supported by several additional lines of evidence. egl-19 encodes the only L-type calcium channel alpha1-subunit in the C. elegans genome. We show that nemadipine-A can also antagonize vertebrate L-type calcium channels, demonstrating that worms and vertebrates share the orthologous protein target. Conversely, FDA-approved DHPs fail to elicit robust phenotypes, making nemadipine-A a unique tool to screen for genetic interactions with this important class of drugs. Finally, we demonstrate the utility of nemadipine-A by using it to reveal redundancy among three calcium channels in the egg-laying circuit. Our study demonstrates that C. elegans enables rapid identification of new small-molecule tools and their targets.

Animals↗
PubMed2005

Stabilin-1 and stabilin-2 are both directed into the early endocytic pathway in hepatic sinusoidal endothelium via interactions with clathrin/AP-2, independent of ligand binding.

Liver sinusoidal endothelial cells (LSECs) mediate clearance of hyaluronan (HA) and scavenger receptor ligands, for example, advanced glycation end product (AGE)-modified proteins and oxidized lipids from the circulation. We recently cloned stabilin-1 and -2, two members of a novel family of transmembrane proteins expressed in LSECs. By using primary LSECs and HEK293 cells separately expressing either stabilin, we have investigated their roles in the early events of endocytosis with respect to localization, ligand-binding properties, and associations with clathrin and adaptor protein (AP)-2. Both stabilins were present at the cell surface, although surface levels of stabilin-1 were limited. In addition, stabilins were present in early endosomal antigen (EEA)-1+ organelles colocalizing with endocytosed AGE-modified bovine serum albumin (BSA). Treating cells with monensin further pronounced this distribution. Recombinant stabilin-2, but not recombinant stabilin-1, bound HA and the scavenger receptor ligands AGE-modified BSA, formaldehyde-treated BSA, and collagen N-terminal propeptides. In LSECs, both stabilins were associated with clathrin and AP-2, but not with each other. These interactions did not change upon addition of exogenous HA, suggesting that stabilins are constitutively internalized. In conclusion, hepatic stabilins are both present in the early endocytic pathway, associating with clathrin/AP-2, but whereas stabilin-2 has a clear scavenging profile, stabilin-1 does not recognize these ligands.

Amino Acid Sequence↗
PubMed2005

Preventing leaf identity theft with hormones.

Genetic analysis of plant development has begun to demonstrate the importance of hormone synthesis and transport in regulating morphogenesis. In the case of leaf development, for example, auxin pooling determines where a primordium will emerge and leads to the activation of transcription factors, which determine leaf identities by modulating abscisic acid (ABA) and gibberellic acid (GA) concentrations. Signal transduction studies suggest that negative regulation of transcription factors through protein turnover is commonly used as a mechanism of hormone action. Together, these findings suggest that auxin might degrade a repressor that allows the activation of genes that modulate ABA/GA ratios in emerging leaves. With our increased understanding of the molecular basis of hormone signaling, it is becoming possible to overlay important regulators onto signaling modules that determine morphological outputs.

Abscisic Acid↗
PubMed2005

Molecular tailoring of farnesylation for plant drought tolerance and yield protection.

Protecting crop yield under drought stress is a major challenge for modern agriculture. One biotechnological target for improving plant drought tolerance is the genetic manipulation of the stress response to the hormone abscisic acid (ABA). Previous genetic studies have implicated the involvement of the beta-subunit of Arabidopsis farnesyltransferase (ERA1) in the regulation of ABA sensing and drought tolerance. Here we show that molecular manipulation of protein farnesylation in Arabidopsis, through downregulation of either the alpha- or beta-subunit of farnesyltransferase enhances the plant's response to ABA and drought tolerance. To test the effectiveness of tailoring farnesylation in a crop plant, transgenic Brassica napus carrying an ERA1 antisense construct driven by a drought-inducible rd29A promoter was examined. In comparison with the non-transgenic control, transgenic canola showed enhanced ABA sensitivity, as well as significant reduction in stomatal conductance and water transpiration under drought stress conditions. The antisense downregulation of canola farnesyltransferase for drought tolerance is a conditional and reversible process, which depends on the amount of available water in the soil. Furthermore, transgenic plants were more resistant to water deficit-induced seed abortion during flowering. Results from three consecutive years of field trial studies suggest that with adequate water, transgenic canola plants produced the same amount of seed as the parental control. However, under moderate drought stress conditions at flowering, the seed yields of transgenic canola were significantly higher than the control. Using protein farnesyltransferase as an effective target, these results represent a successful demonstration of engineered drought tolerance and yield protection in a crop plant under laboratory and field conditions.

Abscisic Acid↗
PubMed2004

The transcription factor FUSCA3 controls developmental timing in Arabidopsis through the hormones gibberellin and abscisic acid.

Although plants continually produce different organs throughout their life cycle, little is known about the factors that regulate the timing of a given developmental program. Here we report that the restricted expression of FUS3 to the epidermis is sufficient to control foliar organ identity in Arabidopsis by regulating the synthesis of two hormones, abscisic acid and gibberellin. These hormones in turn regulate the rates of cell cycling during organ formation to determine whether an embryonic or adult leaf will emerge. We also show that FUS3 expression is influenced by the patterning hormone, auxin, and therefore acts as a nexus of hormone action during embryogenesis. The identification of lipophillic hormones downstream of a heterochronic regulator in Arabidopsis has parallels to mechanisms of developmental timing in animals and suggests a common logic for temporal control of developmental programs between these two kingdoms.

Abscisic Acid↗
PubMed2004

Systems approaches to understanding cell signaling and gene regulation.

The age of 'omics' is upon us, and scientific papers that reflect this are starting to appear at an ever-increasing rate. The amount of information generated in any 'omics' program is daunting and often overwhelms plant scientists whose main interests relate to cell or developmental biology. For this revolution in data generation to have any impact in plant signaling studies, we must have great confidence in both the quality of the data and our ability to represent it in ways that are meaningful to general plant biologists. Systems biology has begun to address these issues and to provide examples in which the analysis of large data sets has led to biological insights into cell signaling and gene regulation.

Arabidopsis↗
PubMed2004

The FUS3 transcription factor functions through the epidermal regulator TTG1 during embryogenesis in Arabidopsis.

Loss-of-function mutations in the FUSCA3 (FUS3) gene of Arabidopsis result in alterations in cotyledon identity, inability to complete late seed maturation processes, and the premature activation of apical and root embryonic meristems, which indicates that this transcription factor is an essential regulator of embryogenesis. Although FUS3 shows a complex pattern of expression in the embryo, this gene is only required in the protoderm to carry out its functions. Moreover, the epidermal morphogenesis regulator TRANSPARENT TESTA GLABRA1 (TTG1) is negatively regulated by FUS3 in the embryo. When a loss-of-function ttg1 mutation is introduced into a fus3 mutant, a number of fus3-related phenotypes are rescued, indicating a functional TTG1 gene is required to manifest the fus3 mutant phenotype. It therefore appears that one of the functions of FUS3 is to restrict the domain of expression of TTG1 during embryogenesis. The FUS3-TTG1 interaction is both maternal and zygotic, suggesting a complex relationship is required between these gene products to allow correct seed development.

Anthocyanins↗
PubMed2003

Hormone signalling from a developmental context.

The influence of hormones on plant growth and development has been clearly documented over the past 50 years. Now, with molecular genetics, the genes that convert changes in hormone levels into a cellular response are beginning to be identified. However, recent studies have demonstrated that the developmental context in which the hormones act plays a large influence on their synthesis and action. In this review, examples are given where known hormone response genes have been shown to have broader developmental roles as well as examples where genes that regulate developmental decisions, such as differentiation and fate, also influence hormone metabolism. The early conclusion of these studies is that an understanding of hormone signal transduction cannot be achieved in the absence of a developmental framework.

Plant Development↗
PubMed2003

The ABSCISIC ACID INSENSITIVE 3 (ABI3) gene is modulated by farnesylation and is involved in auxin signaling and lateral root development in Arabidopsis.

Genetic screens have identified a number of genes that regulate abscisic acid (ABA) responsiveness in Arabidopsis. Using a combination of suppressor screens and double mutant analysis, we have determined a genetic relationship for a number of these ABA response loci. Based on germination in the presence of exogenous ABA, the ABI1 and ABI2 phosphatases act at or upstream of the ERA1 farnesyl transferase and the ABI3 and ABI5 transcription factors act at or downstream of ERA1. In contrast with ABI3 and ABI5, the ABI4 transcription factor appears to act at or upstream of ERA1. Based on reporter gene constructs, the upstream regulation of ABI3 by ERA1 occurs at least partially at the level of transcription, suggesting that this lipid modification is required to attenuate ABI3 expression. Similar experiments also indicate that ABI3 is auxin inducible in lateral root primordia. Related to this, loss-of-function abi3 alleles show reduced lateral root responsiveness in the presence of auxin and an auxin transport inhibitor, and era1 mutants have increased numbers of lateral roots. These results suggest the possibility that genes identified through ABA responsive germination screens such as ERA1 and ABI3 have functions in auxin action in Arabidopsis.

Abscisic Acid↗
PubMed2003

Cross-talk in plant hormone signalling: what Arabidopsis mutants are telling us.

Genetic screens have been extremely useful in identifying genes involved in hormone signal transduction. However, although these screens were originally designed to identify specific components involved in early hormone signalling, mutations in these genes often confer changes in sensitivity to more than one hormone at the whole-plant level. Moreover, a variety of hormone response genes has been identified through screens that were originally designed to uncover regulators of sugar metabolism. Together, these facts indicate that the linear representation of the hormone signalling pathways controlling a specific aspect of plant growth and development is not sufficient, and that hormones interact with each other and with a variety of developmental and metabolic signals. Following the advent of arabidopsis molecular genetics we are beginning to understand some of the mechanisms by which a hormone is transduced into a cellular response. In this Botanical Briefing we review a subset of genes in arabidopsis that influence hormonal cross-talk, with emphasis on the gibberellin, abscisic acid and ethylene pathways. In some cases it appears that modulation of hormone sensitivity can cause changes in the synthesis of an unrelated hormone, while in other cases a hormone response gene defines a node of interaction between two response pathways. It also appears that a variety of hormones may converge to regulate the turnover of important regulators involved in growth and development. Examples are cited of the recent use of suppressor and enhancer analysis to identify new nodes of interaction between these pathways.

Abscisic Acid↗
PubMed2002

A screen for genes that function in abscisic acid signaling in Arabidopsis thaliana.

The plant hormone abscisic acid (ABA) controls many aspects of plant growth and development under a diverse range of environmental conditions. To identify genes functioning in ABA signaling, we have carried out a screen for mutants that takes advantage of the ability of wild-type Arabidopsis seeds to respond to (-)-(R)-ABA, an enantiomer of the natural (+)-(S)-ABA. The premise of the screen was to identify mutations that preferentially alter their germination response in the presence of one stereoisomer vs. the other. Twenty-six mutants were identified and genetic analysis on 23 lines defines two new loci, designated CHOTTO1 and CHOTTO2, and a collection of new mutant alleles of the ABA-insensitive genes, ABI3, ABI4, and ABI5. The abi5 alleles are less sensitive to (+)-ABA than to (-)-ABA. In contrast, the abi3 alleles exhibit a variety of differences in response to the ABA isomers. Genetic and molecular analysis of these alleles suggests that the ABI3 transcription factor may perceive multiple ABA signals.

Abscisic Acid↗
PubMed1999

GENETIC ANALYSIS OF HORMONE SIGNALING.

Phytohormones influence many diverse developmental processes ranging from seed germination to root, shoot, and flower formation. Recently, mutational analysis using the model plant Arabidopsis thaliana has been instrumental in determining the individual components of specific hormone signal transduction pathways. Moreover, epistasis and suppressor studies are beginning to explain how these genes and their products relate to one another. While no hormone transduction pathway is completely understood, the genes identified to date suggest that simple molecular rules can be established to explain how plant hormone signals are transduced. This review describes some of the shared characteristics of plant hormone signal transduction pathways and the properties for informational transfer common to many of the genes that specify the transduction of the signal.

Journal Article↗