Light-induced effects of several enzymes of carbohydrate metabolism in Phycomyces blakesleeanus

• 1.1. The photoregulation shown by glyceraldehyde 3-phosphate dehydrogenase and glucose 6-phosphate dehydrogenase appears to be independent of the mad gene product(s) and also independent of carotene biosynthesis regulation.
• 2.2. The photoregulation of malate dehydrogenase appeared to be dependent on the mutation of the mad and car S genes.
• 3.3. Pyruvate kinase and lactate dehydrogenase may be classified as light-independent.
• 4.4. The action of ATP and fructose 1,6-bisphosphate on the enzymes studied was generally independent of light/dark grown conditions.
• 5.5. However, the effect of fructose 1,6-bisphosphate on Phycomyces pyruvate kinase appears to be light-dependent.

     

Photoregulation of morphogenesis of tobacco plants transformed with the gene of human interlenkin-18

The effects of blue light (BL), green light (GL), and red light (RL) on morphogenesis photoregulation of transgenic tobacco (Nicotiana tabacum L.) plants containing a copy of human interleukin-18 gene were studied. Wild-type and transgenic (Il1 No.87–1 and Il18 No.7–11) lines and derived calluses were used. Photomorphogenesis of transgenic lines under GL and RL differed substantially from that of initial line at early stages of morphogenesis; this was expressed in hypocotyl lengthening under GL and cotyledon enhanced expansion under RL. Growth responses to light quality were related to changes in the levels of zeatin, IAA, and ABA. Thus, hypocotyl lengthening and increased cotyledon area under GL occurred at the elevated level of IAA and reduced level of ABA. Growth of callus cells in transgenic lines during zero passage differed from wild-type calluses only in darkness. The data obtained indicate that tobacco plant transformation with the human interleukin-18 gene changed functioning of the photoregulation systems at early developmental stages. This phenomenon is evidently explained by the pleiotropic effects of the gene inserted.     

Differential usage of photoreceptors for chalcone synthase gene expression during plant development

Photoregulation of chalcone synthase (CHS) mRNA accumulation was analysed in parsley (Petroselinum crispum) and mustard (Sinapis alba) plants at different developmental stages. In both species, CHS mRNA accumulation in young etiolated seedlings was primarily under phytochrome control. In leaves of adult re-etiolated plants, a UV-B photoreceptor was predominantly involved in photocontrol. The reduced red light control in mature leaves was not due to the absence of immunoreactive phytochrome. The apparent dependence of photoreceptor usage on the developmental state of the cell or organism was in accordance with observations on the photoregulation of fusion constructs between CHS promoters from parsley or mustard and the β-glucuronidase reporter gene (GUS). When tested in the parsley protoplast transient expression system, both constructs yielded the same type of photoregulation as observed for the endogenous CHS gene.     

Photoregulation of cellular morphology during complementary chromatic adaptation requires sensor-kinase-class protein RcaE in Fremyella diplosiphon

We used wild-type UTEX481; SF33, a shortened-filament mutant strain that shows normal complementary chromatic adaptation pigmentation responses; and FdBk14, an RcaE-deficient strain that lacks light-dependent pigmentation responses, to investigate the molecular basis of the photoregulation of cellular morphology in the cyanobacterium Fremyella diplosiphon. Detailed microscopic and biochemical analyses indicate that RcaE is required for the photoregulation of cell and filament morphologies of F. diplosiphon in response to red and green light.     

Oleic acid metabolism: A biochemical marker for studying photoregulation of epicotyl elongation in Vigna unguiculata

Oleic acid metabolism can be considered to be an indicator of growth photoregulation in cowpea ( Vigna unguiculata Westphal cv. M53) epicotyls. Phosphatidylcholine and phosphatidylethanolamine are the two lipid classes concerned in the photoregulation of oleic acid accumulation. The incorporation of radioactive precursors in internodes of whole plants has shown that there is de novo synthesis of these phospholipids during the light dependent growth process.
The variations in oleic acid content were used to study the photocontrol of elongation in segments excised from the apical part of the epicotyl. In this system, as in whole plants, oleic acid was the only fatty acid showing significant variation related to the light/dark treatments. Differences in photoresponse between excised intenode segments and internodes in whole plants are discussed.     

Photoresponsive 5'-cap for the reversible photoregulation of gene expression

Photoresponsive 5'-caps that can be reversibly cis-trans isomerized by light irradiation were developed for the reversible photoregulation of gene expression. The 8-naphthylvinyl cap (8NV-cap) in the trans form completely inhibited translation of mRNA, whereas the cis form yielded protein with the same efficiency as mRNA capped with the normal-cap, a 26-fold higher efficiency than that of the trans form. The 8NV-capped mRNA could be switched between a translating state (ON state) and a non-translating state (OFF state) in a reversible fashion by alternately irradiating with monochromatic 410 nm or 310 nm light.     

Spatial-specific regulation of root development by phytochromes in Arabidopsis thaliana

Distinct tissues and organs of plants exhibit dissimilar responses to light exposure – cotyledon growth is promoted by light, whereas hypocotyl growth is inhibited by light. Light can have different impacts on root development, including impacting root elongation, morphology, lateral root proliferation and root tropisms. In many cases, light inhibits root elongation. There has been much attention given to whether roots themselves are the sites of photoperception for light that impacts light-dependent growth and development of roots. A number of approaches including photoreceptor localization in planta, localized irradiation and exposure of dissected roots to light have been used to explore the site(s) of light perception for the photoregulation of root development. Such approaches have led to the observation that photoreceptors are localized to roots in many plant species, and that roots are capable of light absorption that can alter morphology and/or gene expression. Our recent results show that localized depletion of phytochrome photoreceptors in Arabidopsis thaliana disrupts root development and root responsiveness to the plant hormone jasmonic acid. Thus, root-localized light perception appears central to organ-specific, photoregulation of growth and development in roots.     

Downstream effectors of light- and phytochrome-dependent regulation of hypocotyl elongation in Arabidopsis thaliana

Arabidopsis, like most plants, exhibits tissue-specific, light-dependent growth responses. Cotyledon and leaf growth and the accumulation of photosynthetic pigments are promoted by light, whereas hypocotyl growth is inhibited. The identification and characterization of distinct phytochrome-dependent molecular effectors that are associated with these divergent tissue-specific, light-dependent growth responses are limited. To identify phytochrome-dependent factors that impact the photoregulation of hypocotyl length, we conducted comparative gene expression studies using Arabidopsis lines exhibiting distinct patterns of phytochrome chromophore inactivation and associated disparate hypocotyl elongation responses under far-red (FR) light. A large number of genes was misregulated in plants lacking mesophyll-specific phytochromes relative to constitutively-deficient phytochrome lines. We identified and characterized genes whose expression is impacted by light and by phyA and phyB that have roles in the photoregulation of hypocotyl length. We characterized the functions of several identified target genes by phenotyping of T-DNA mutants. Among these genes is a previously uncharacterized LHE (LIGHT-INDUCED HYPOCOTYL ELONGATION) gene, which we show impacts light- and phytochrome-mediated regulation of hypocotyl elongation under red (R) and FR illumination. We describe a new approach for identifying genes involved in light- and phytochrome-dependent, tissue-specific growth regulation and confirmed the roles of three such genes in the phytochrome-dependent photoregulation of hypocotyl length.     

Lesions in many different spindle components activate the spindle checkpoint in the budding yeast Saccharomyces cerevisiae.

The spindle checkpoint arrests cells in mitosis in response to defects in the assembly of the mitotic spindle or errors in chromosome alignment. We determined which spindle defects the checkpoint can detect by examining the interaction of mutations that compromise the checkpoint (mad1, mad2, and mad3) with those that damage various structural components of the spindle. Defects in microtubule polymerization, spindle pole body duplication, microtubule motors, and kinetochore components all activate the MAD-dependent checkpoint. In contrast, the cell cycle arrest caused by mutations that induce DNA damage (cdc13), inactivate the cyclin proteolysis machinery (cdc16 and cdc23), or arrest cells in anaphase (cdc15) is independent of the spindle checkpoint.     

Phytochrome regulation of nuclear gene expression in plants

The photoregulation of gene expression in higher plants was extensively studied during the 1980s, in particular the light-responsive cis -acting elements and trans -acting factors of the Lhcb and rbcS genes. However, little has been discovered about: (1) which plant genes are regulated by light, and (2) which photoreceptors control the expression of these genes. In the 1990s, the functional analysis of the various photoreceptors has progressed rapidly using photoreceptor-deficient mutants, including those of the phytochrome gene family. More recently however, advanced techniques for gene expression analysis, such as fluorescent differential display and DNA microarray technology, have become available enabling the global identification of genes that are regulated by particular photoreceptors. In this paper we describe distinct and overlapping effects of individual phytochromes on gene expression in Arabidopsis thaliana.     

Feedback control of mitosis in budding yeast.

We have investigated the feedback control that prevents cells with incompletely assembled spindles from leaving mitosis. We isolated budding yeast mutants sensitive to the anti-microtubule drug benomyl. Mitotic arrest-deficient (mad) mutants are the subclass of benomyl-sensitive mutants in which the completion of mitosis is not delayed in the presence of benomyl and that die as a consequence of their premature exit from mitosis. A number of properties of the mad mutants indicate that they are defective in the feedback control over the exit from mitosis: their killing by benomyl requires passage through mitosis; their benomyl sensitivity can be suppressed by an independent method for delaying the exit from mitosis; they have normal microtubules; and they have increased frequencies of chromosome loss. We cloned MAD2, which encodes a putative calcium-binding protein whose disruption is lethal. We discuss the role of feedback controls in coordinating events in the cell cycle.     

Photoregulatory behavior of bluegill,Lepomis macrochira,in a virtual light gradient

Synopsis Vertical movements of bluegill were monitored in gradients of light intensity to assess this fish's photoregulatory ability and mechanisms. A computerized monitoring and control system created virtual gradients of light intensity by adjusting an overhead lamp's output in response to fish movements, in a vertical tube, to produce a programmed intensity at the fish's depth position. This approach separated the process of gradient formation from normal clues for photoregulation and allowed formation of light gradients incompatible with natural taxic responses to intensity. Hourly shifts in gradient position minimized the possibility of confounding photoregulation with position regulation. Observed patterns of movement reduced the extremes of light intensity to which bluegill were exposed, compared to no movement or random movement. Seven fish were tested, producing 10 experiments. In 4 of 10 experiments, the fish effectively photoregulated in gradients in which light intensity decreased with depth, as in natural habitats. In 1 of 10 experiments, the fish photoregulated in an inverse gradient, with intensity increasing with depth. Evidence of regulation in an inverse gradient suggests that normal taxic responses are not essential for photoregulation in bluegill.     

The Spindle Checkpoint of Budding Yeast Depends on a Tight Complex between the Mad1 and Mad2 Proteins

The spindle checkpoint arrests the cell cycle at metaphase in the presence of defects in the mitotic spindle or in the attachment of chromosomes to the spindle. When spindle assembly is disrupted, the budding yeast mad and bub mutants fail to arrest and rapidly lose viability. We have cloned the MAD2 gene, which encodes a protein of 196 amino acids that remains at a constant level during the cell cycle. Gel filtration and co-immunoprecipitation analyses reveal that Mad2p tightly associates with another spindle checkpoint component, Mad1p. This association is independent of cell cycle stage and the presence or absence of other known checkpoint proteins. In addition, Mad2p binds to all of the different phosphorylated isoforms of Mad1p that can be resolved on SDS-PAGE. Deletion and mutational analysis of both proteins indicate that association of Mad2p with Mad1p is critical for checkpoint function and for hyperphosphorylation of Mad1p.     

Photoregulation of Protochlorophyllide Oxidoreductase and Rubisco Large Subunit Accumulation in Phytochrome A-Deficient Transgenic Tobacco Plants

Some morphogenetic responses, induced by far red (FR) light in tobacco plants (Nicotiana tabacum L.), were studied. The inhibitory effect of FR irradiation on chlorophyll synthesis in transgenic plants with reduced phytochrome A content was almost absent. Phytochrome A-mediated repression of the por gene was demonstrated with the use of polyclonal antiserum against protochlorophyllide oxidoreductase. Continuous FR light induced the accumulation of Rubisco large subunits in wild-type but not in transgenic tobacco plants. Our data confirm the suggestion that phytochrome A mediates photoregulation of the synthesis of these proteins.     

C-terminal region of Mad2 plays an important role during mitotic spindle checkpoint in fission yeast <Emphasis Type="Italic">S</Emphasis>c<Emphasis Type="Italic">hizosaccharomyces pombe</Emphasis>

The mitotic arrest deficiency 2 (Mad2) protein is an essential component of the spindle assembly checkpoint that interacts with Cdc20/Slp1 and inhibit its ability to activate anaphase promoting complex/cyclosome (APC/C). In bladder cancer cell line the C-terminal residue of the mad2 gene has been found to be deleted. In this study we tried to understand the role of the C-terminal region of mad2 on the spindle checkpoint function. To envisage the role of C-terminal region of Mad2, we truncated 25 residues of Mad2 C-terminal region in fission yeast S.pombe and characterized its effect on spindle assembly checkpoint function. The cells containing C-terminal truncation of Mad2 exhibit sensitivity towards microtubule destabilizing agent suggesting perturbation of spindle assembly checkpoint. Further, the C-terminal truncation of Mad2 exhibit reduced viability in the nda3-KM311 mutant background at non-permissive temperature. Truncation in mad2 gene also affects its foci forming ability at unattached kinetochore suggesting that the mad2-?CT mutant is unable to maintain spindle checkpoint activation. However, in response to the defective microtubule, only brief delay of mitotic progression was observed in Mad2 C-terminal truncation mutant. In addition we have shown that the deletion of two β strands of Mad2 protein abolishes its ability to interact with APC activator protein Slp1/Cdc20. We purpose that the truncation of two β strands (β7 and β8) of Mad2 destabilize the safety belt and affect the Cdc20-Mad2 interaction leading to defects in the spindle checkpoint activation.