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PubMed2026

Integrated metabolomic, transcriptomic, and proteomic analyses reveal changes in the non-volatile metabolite profile of LED light-withered oolong tea.

LED light withering is a crucial method for overcoming weather limitations and enhancing the quality of oolong tea. To elucidate the underlying molecular mechanisms, this study simulated solar spectra using multiwavelength LED light and compared the resulting metabolic, transcriptomic, and proteomic profiles during the enzymatic-catalysis process (ECP) in oolong tea processing. Results indicated that LED light withering altered gene expression and protein regulation of secondary metabolism, particularly in the flavonoid biosynthesis pathway. These shifts encompassed key quality-related compounds, including flavonoids (quercetin-3-O-rhamnoside, dihydroquercetin), amino acids (L-asparagine, L-histidine), guanosine 5'-monophosphate (GMP), and carbohydrates. Furthermore, LED light withering accelerated tea leaf water loss, influenced gene expression involved in photosynthetic cellular components (chloroplasts, thylakoids), increased ascorbate peroxidase regulation under stress, and subsequently modulated energy metabolism and signal transduction in tea leaves. This study offers molecular theoretical framework for the controlled light-withering of oolong tea under bad weather and the associated improvements in its quality.

Camellia sinensis↗
PubMed1994

[Mechanisms of bronchial hyperreactivity. Role of smooth muscle].

The understanding of the cellular and molecular mechanisms involved in the contraction of airway smooth muscle has tremendously advanced in the recent years. Extracellular messengers that control the muscle contraction as well as both the membrane transduction processes and the activation of the contractile proteins are known in some details. Sensitization alters the contractility of airway smooth muscle. It increases shortening in the early phase of the contraction when rapidly cycling cross bridges are operative. At the molecular level, the responsible mechanism is an increment in the activity of the processes leading to phosphorylation of the myosin light chain. Such an alteration in the airway smooth muscle contractility may be observed in the early phase of the development of bronchial hyperresponsiveness.

Animals↗
PubMed1999

Arabidopsis thaliana 'extra-large GTP-binding protein' (AtXLG1): a new class of G-protein.

Heterotrimeric GTP-binding proteins, composed of alpha, beta, and gamma subunits, are involved in signal transduction pathways in animal and plant systems. In plants, physiological analyses implicate heterotrimeric G-proteins in ion channel regulation, light signaling, and hormone and pathogen responses. However, only one class of plant G alpha genes has been identified to date. We have cloned a novel gene, 'Arabidopsis thaliana extra-large GTP-binding protein' (AtXLG1). AtXLG1 appears to be a member of a small gene family and is transcribed in all tissues assayed: roots, leaves, stems, flowers, and fruits. The conceptually translated protein from AtXLG1 is 99 kDa, twice as large as typical G alpha proteins. The carboxy-terminal half of the AtXLG1 protein has significant homology to animal and plant G alpha proteins. This region includes a GTP-binding domain, a predicted helical domain, and an aspartate/glutamate-rich loop, which are characteristics of G alpha's. Despite the absence of some of the amino acids implicated in GTP binding and hydrolysis by crystallographic and mutational analyses of mammalian G alpha's, recombinant AtXLG1 binds GTP with specificity. The amino-terminal region of AtXLG1 contains domains homologous to the bacterial TonB-box, which is involved in energy transduction between the inner and outer bacterial membranes, and to zinc-finger proteins. Given the unique structure of AtXLG1, it will be of interest to uncover its physiological functions.

Amino Acid Sequence↗
PubMedDate not supplied

[Photoreceptor apparatus of a fungus Neurospora crassa].

Light governs living functions of ascomycete fungus Neurospora crassa by controlling expression of the genes responsible for differentiation of reproductive structures, synthesis of secondary metabolites and the circadian oscillator activity. Illumination also influences electrogenic processes in cell membrane and the activity and molecular organization of some enzymes. The major but, probably, not the sole photoreceptor pigment in Neurospora cells is WCC, a heterodimeric complex formed by PAS-domain-containing polypeptides WC-1 and WC-2, which are the products of genes white collar-1 and white collar-2. Mutation of any of these genes arrests most of the organism's responses to light. The photoreceptor belongs to a recently discovered vast group of non-homologous light-sensitive proteins, whose molecules bind flavin coenzymes as the photosensor chromophores. The photosignal transduction is started by excitation and photochemical activity of excited FAD molecule non-covalently bound by LOV-domain (a specialized version of PAS-domain) in WC-1 protein. The presence in both WC-1- and WC-2-proteins of "zinc fingers" (the GATA recognizing sequences) suggested that these motives might act as transcription factors. The critical analysis of photoinduction mechanism has shown, however, that promoters of light-sensitive genes do not contain a common cis-acting element, what makes to look for alternative mechanisms underlying photoregulated gene activity.

DNA-Binding Proteins↗
PubMed1993

Inhibitory effects of KN-93, an inhibitor of Ca2+ calmodulin-dependent protein kinase II, on light-regulated root gravitropism in maize.

Light is essential for root gravitropism in Zea mays L., cultivar Merit. It is hypothesized that calcium mediates this light-regulated response. KN-93, an inhibitor of calcium/calmodulin kinase II (CaMK II), inhibits light-regulated root gravitropism but does not affect light perception. We hypothesize that CaMK II, or a homologue, operates late in the light/gravity signal transduction chain. Here we provide evidence suggesting a possible physiological involvement of CaMK II in root gravitropism in plants.

Benzylamines↗
PubMed1990

A new gene (madI) involved in the phototropic response of Phycomyces.

Only eight genes are known to be involved in the phototropic response of Phycomyces (madA-H). Mutants affected in these genes have played a major role in the analysis of photosensory transduction processes in this system. A set of new mutants isolated by Alvarez et al. (1989) that are unable to bend towards dim unilateral blue light were studied by complementation and recombination. Two of these mutants have mutations in madE, one has a mutation in madF and one is a double madE madF mutant. The three remaining mutants tested did not complement each other and showed positive complementation with strains carrying mutations in the genes madA, madB, and madC, indicating that they carried mutations in a new gene designated madI. Recombination analysis showed that madI is unlinked to madA, madB and madC.

Genes, Fungal↗
PubMed2003

Growth suppression, altered stomatal responses, and augmented induction of heat shock proteins in cytosolic ascorbate peroxidase (Apx1)-deficient Arabidopsis plants.

The accumulation of hydrogen peroxide (H2O2) in plants is typically associated with biotic or abiotic stresses. However, H2O2 is continuously produced in cells during normal metabolism. Yet, little is known about how H2O2 accumulation will affect plant metabolism in the absence of pathogens or abiotic stress. Here, we report that a deficiency in the H2O2-scavenging enzyme, cytosolic ascorbate peroxidase (APX1), results in the accumulation of H2O2 in Arabidopsis plants grown under optimal conditions. Knockout-Apx1 plants were characterized by suppressed growth and development, altered stomatal responses, and augmented induction of heat shock proteins during light stress. The inactivation of Apx1 resulted in the induction of several transcripts encoding signal transduction proteins. These were not previously linked to H2O2 signaling during stress and may belong to a signal transduction pathway specifically involved in H2O2 sensing during normal metabolism. Surprisingly, the expression of transcripts encoding H2O2 scavenging enzymes, such as catalase or glutathione peroxidase, was not elevated in knockout-Apx1 plants. The expression of catalase, two typical plant peroxidases, and several different heat shock proteins was however elevated in knockout-Apx1 plants during light stress. Our results demonstrate that in planta accumulation of H2O2 can suppress plant growth and development, interfere with different physiological processes, and enhance the response of plants to abiotic stress conditions. Our findings also suggest that at least part of the induction of heat shock proteins during light stress in Arabidopsis is mediated by H2O2 that is scavenged by APX1.

Arabidopsis↗
PubMedDate not supplied

A phytochrome-like protein AphC triggers the cAMP signaling induced by far-red light in the cyanobacterium Anabaena sp. strain PCC7120.

In the filamentous, nitrogen-fixing cyanobacterium Anabaena sp. PCC7120, red light (630 nm) decreased, whereas far-red light (720 nm) increased cellular adenosine 3',5'-cyclic monophosphate (cAMP) content. To find a red and far-red light photoreceptor that triggers the cAMP signal cascade, we disrupted 10 open reading frame having putative chromophore-binding GAF domains. The response of the cellular cAMP concentration to red and far-red light in each open reading frame disruptant was determined. It was found that only the mutant of the gene all2699 failed to respond to far-red light. The open reading frame named as aphC encoded a protein with 920 amino acids including GAF domains similar to those involved in Cph2, a photoreceptor of Synechocystis sp. PCC6803. To determine which adenylate cyclase (AC) is responsible for far-red light signal, we disrupted all AC genes and found that CyaC was the candidate. The enzymatic activity of CyaC might be controlled by a far-red light photoreceptor through the phosphotransfer reaction. The site-specific mutant of the Asp59 residue of the receiver (R1) domain of CyaC lost its light-response capability. It was suggested that the far-red light signal was received by AphC and then transferred to the N-terminal response regulator domain of CyaC. Then its catalytic activity was stimulated, which increased the cellular cAMP concentration and drove the subsequent signal transduction cascade.

Adenylyl Cyclases↗
PubMed2003

Localization of the two constitutively expressed nitric oxide synthase isoforms (nNOS and eNOS) in the same cell types in the saccule maculae of the frog Rana pipiens by immunoelectron microscopy: evidence for a back-up system?

There is growing evidence for a nitric oxide/cyclic GMP pathway of signal transduction in the vestibular system. Recently, two isoforms of nitric oxide (NO) synthase (nNOS and eNOS) and NO itself have been identified at the light microscopic level in the vestibulocochlear system of mice using specific antibodies and a new fluorescence indicator. In order to acquire more information about signal transduction and tissue modulation in this neuroepithelium at the cellular and subcellular levels, ultrathin sections of London Resin White-embedded saccule maculae of the frog Rana pipiens were incubated with various concentrations of commercially available antibodies to nNOS and eNOS. The immunoreactivity was visualized by a gold-labelled secondary antibody and the amount of the immunoreactions per microm2 was quantified for the different cell types and subcellular regions. Significant eNOS immunoreactivity was identified in the hair bundles, cuticular plates and the rest of the cytoplasm of the hair cells as well as in different subcellular regions of the supporting cells. Gold-labelled anti-nNOS antibodies stained mainly stereovilli and cuticular structures of hair cells and supporting cells, whereas the number of the immunoreactions in the remaining cytoplasm of both cell types was near the background level. The spatial co-localization of the two NOS isotypes in the same cell regions of hair cells and supporting cells was confirmed in double-labelling experiments. The immunocytochemical findings are suggestive of a redundant system in which one NOS isoform can (partially) replace the other. The different subcellular localization of the NOS isoforms may allow for isoform specific regulation of NOS activity by different Ca2+ currents at the subcellular level, underlining the importance of NO-regulated processes in neuroepithelia of the inner ear.

Acoustic Maculae↗
PubMed2004

Plant body weight-induced secondary growth in Arabidopsis and its transcription phenotype revealed by whole-transcriptome profiling.

Wood is an important raw material and environmentally cost-effective renewable source of energy. However, the molecular biology of wood formation (i.e. secondary growth) is surprisingly understudied. A novel experimental system was employed to study the molecular regulation of secondary xylem formation in Arabidopsis. First, we demonstrate that the weight carried by the stem is a primary signal for the induction of cambium differentiation and the plant hormone, auxin, is a downstream carrier of the signal for this process. We used Arabidopsis whole-transcriptome (23 K) GeneChip analysis to examine gene expression profile changes in the inflorescent stems treated for wood formation by cultural manipulation or artificial weight application. Many of the genes up-regulated in wood-forming stems had auxin responsive cis-acting elements in their promoter region, indicating auxin-mediated regulation of secondary growth. We identified 700 genes that were differentially expressed during the transition from primary growth to secondary growth. More than 40% of the genes that were up-regulated (>5x) were associated with signal transduction and transcriptional regulation. Biological significance of these regulatory genes is discussed in light of the induction and development of secondary xylem.

Arabidopsis↗
PubMed2005

Proteins in action monitored by time-resolved FTIR spectroscopy.

In the post genome era proteins coming into the focus of life sciences. X-ray structure analysis and NMR spectroscopy are established methods to determine the geometry of proteins. In order to determine the molecular reaction mechanism of proteins, time-resolved FTIR (trFTIR) difference spectroscopy emerges as a valuable tool. In this Minireview we describe the trFTIR difference spectroscopy and show its application on the light-driven proton pump bacteriorhodopsin (bR), the photosynthetic reaction center and the GTPase Ras, which is crucial in signal transduction. The main principles of the technique are presented, including a summary of triggering techniques, scan modes and analysis.

Bacteriorhodopsins↗
PubMed2003

Phototaxis, chemotaxis and the missing link.

Phototaxis in Archaea employs an integral membrane complex composed of a photoreceptor that is similar to the light-driven proton pump bacteriorhodopsin, and a transducer protein that is similar to the familiar eubacterial chemotaxis receptors. Recent structural studies have revealed how these proteins are assembled in the membrane, and provide a heuristic framework for future work on the mechanism of signal transduction by this important class of molecules.

Archaea↗
PubMed1994

Involvement of superoxide radical in signal transduction regulating stomatal movements.

Stomatal movements are regulated by alternate phase transition in the lipophilic domains of membrane initiated by blue light through activation of plasmalemma redox system. ABA blocks the blue light activated redox reactions leading to the accumulation of superoxide (O2.-) radical inducing melting of the membrane and increased fluidity. Phenolic compounds which reverse the ABA effects reduce O2.- radical and thereby reverse phase transition in lipophilic domains of membrane. Stomatal movements thus appear to be regulated by blue light activated redox reactions involving formation of O2.- radical and its reduction by the reducing equivalents available in the GCPs.

Abscisic Acid↗
PubMed2002

Interactions between gravitropism and phototropism in plants.

To receive adequate light and nutrients for survival, plants orient stems and stem-like organs toward light and away from the gravity vector and, conversely, orient roots into the soil, away from light toward the direction of gravity. Therefore, both gravity and light can influence the differential growth of plant organs. To add to the complexity of the interactions between gravity and light, each stimulus can enhance or reduce the effectiveness of the other. On earth, the constant presence of gravity makes it difficult to determine whether plant growth and development is influenced by gravity or light alone or the combination of the two stimuli. In the past decade, our understanding of the gravity and light transduction pathways has advanced through the use of mutants in either gravitropic or phototropic responses and the use of innovative techniques that reduce the effects of one stimulus on the other. Thus, both unique and common elements in the transduction pathways of the gravitropic and phototropic responses have been isolated. This article is focused on the interactions between the light- and gravity-transduction pathways and describes methods used to separate the influences of these two environmental stimuli.

Arabidopsis↗
PubMed1994

Intracellular signal transduction for vasoactive peptides in hypertension.

Increased peripheral resistance is the hallmark of hypertension. It may result in part from exaggerated vascular reactivity of resistance arteries. Some changes in density of surface receptors for different vasoconstrictors and vasorelaxants have been described that could play a role in physiological findings in hypertension. Smooth muscle cells of resistance arteries have increased cytosolic free calcium concentration in some models of experimental hypertension, which may contribute to enhance vascular responses. Exaggerated response of the inositol phosphate-calcium pathway has been demonstrated after stimulation with some vasoconstrictor agents such as norepinephrine, angiotension II, and vasopressin. In contrast, responses to the potent vasoconstrictor peptide endothelin-1 are either normal (in spontaneously hypertensive rats) or blunted (in deoxycorticosterone-salt hypertension). In the latter case, endothelin receptor density, inositol phosphate and diacylglycerol generation, and cytosolic calcium responses agree with blunted response of blood vessels. Increased basal cytosolic calcium and exaggerated sensitivity of myosin light chain to calcium may be mechanisms underlying increases in sensitivity of signal transduction in smooth muscle in some models of hypertension. However, in general, signal transduction of receptors for vasoconstrictors appears to be blunted rather than exaggerated, except for responses to angiotensin II. Altered structure of resistance arteries (remodeling) may be a mechanism that, even in presence of blunted intracellular signal transduction, may result in enhanced pressor responsiveness of blood vessels in hypertension.

Animals↗
PubMed2005

Predicting the signaling state of photoactive yellow protein.

As a bacterial blue light sensor the photoactive yellow protein (PYP) undergoes conformational changes upon signal transduction. The absorption of a photon triggers a series of events that are initially localized around the protein chromophore, extends to encompass the whole protein within microseconds, and leads to the formation of the transient pB signaling state. We study the formation of this signaling state pB by molecular simulation and predict its solution structure. Conventional straightforward molecular dynamics is not able to address this formation process due to the long (microsecond) timescales involved, which are (partially) caused by the presence of free energy barriers between the metastable states. To overcome these barriers, we employed the parallel tempering (or replica exchange) method, thus enabling us to predict qualitatively the formation of the PYP signaling state pB. In contrast to the receptor state pG of PYP, the characteristics of this predicted pB structure include a wide open chromophore-binding pocket, with the chromophore and Glu(46) fully solvent-exposed. In addition, loss of alpha-helical structure occurs, caused by the opening motion of the chromophore-binding pocket and the disruptive interaction of the negatively charged Glu(46) with the backbone atoms in the hydrophobic core of the N-terminal cap. Recent NMR experiments agree very well with these predictions.

Bacterial Proteins↗
PubMed2000

The visual G protein of fly photoreceptors interacts with the PDZ domain assembled INAD signaling complex via direct binding of activated Galpha(q) to phospholipase cbeta.

Visual transduction in the compound eye of flies is a well-established model system for the study of G protein-coupled transduction pathways. Pivotal components of this signaling pathway, including the principal light-activated Ca(2+) channel transient receptor potential, an eye-specific protein kinase C, and the norpA-encoded phospholipase Cbeta, are assembled into a supramolecular signaling complex by the modular PDZ domain protein INAD. We have used immunoprecipitation assays to study the interaction of the heterotrimeric visual G protein with this INAD signaling complex. Light-activated Galpha(q)- guanosine 5'-O-(thiotriphosphate) and AlF(4)(-)-activated Galpha(q), but not Gbetagamma, form a stable complex with the INAD signaling complex. This interaction requires the presence of norpA-encoded phospholipase Cbeta, indicating that phospholipase Cbeta is the target of activated Galpha(q). Our data establish that the INAD signaling complex is a light-activated target of the phototransduction pathway, with Galpha(q) forming a molecular on-off switch that shuttles the visual signal from activated rhodopsin to INAD-linked phospholipase Cbeta.

Animals↗