34 research outputs found

    Protein PII regulates both inorganic carbon and nitrate uptake and is modified by a redox signal in Synechocystis PCC 6803

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    AbstractIn Synechocystis PCC 6803 as in other cyanobacteria, involvement of protein PII in the co-regulation of inorganic carbon and nitrogen metabolism was established based on post-translational modifications of the protein resulting from changes in the carbon/nitrogen regimes. Uptake of bicarbonate and nitrate in response to changes of the carbon and/or nitrogen regimes is altered in a PII-null mutant, indicating that both processes are under control of PII. Modulation of electron flow by addition of methyl viologen with or without duroquinol, or in a NAD(P)H dehydrogenase-deficient mutant, affects the phosphorylation level of PII. The redox state of the cells would thus act as a trigger for PII phosphorylation

    Salt shock-inducible Photosystem I cyclic electron transfer in Synechocystis PCC6803 relies on binding of ferredoxin:NADP+ reductase to the thylakoid membranes via its CpcD phycobilisome-linker homologous N-terminal domain

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    AbstractRelative to ferredoxin:NADP+ reductase (FNR) from chloroplasts, the comparable enzyme in cyanobacteria contains an additional 9 kDa domain at its amino-terminus. The domain is homologous to the phycocyanin associated linker polypeptide CpcD of the light harvesting phycobilisome antennae. The phenotypic consequences of the genetic removal of this domain from the petH gene, which encodes FNR, have been studied in Synechocystis PCC 6803. The in frame deletion of 75 residues at the amino-terminus, rendered chloroplast length FNR enzyme with normal functionality in linear photosynthetic electron transfer. Salt shock correlated with increased abundance of petH mRNA in the wild-type and mutant alike. The truncation stopped salt stress-inducible increase of Photosystem I-dependent cyclic electron flow. Both photoacoustic determination of the storage of energy from Photosystem I specific far-red light, and the re-reduction kinetics of P700+, suggest lack of function of the truncated FNR in the plastoquinone–cytochrome b6f complex reductase step of the PS I-dependent cyclic electron transfer chain. Independent gold-immunodecoration studies and analysis of FNR distribution through activity staining after native polyacrylamide gelelectrophoresis showed that association of FNR with the thylakoid membranes of Synechocystis PCC 6803 requires the presence of the extended amino-terminal domain of the enzyme. The truncated ΔpetH gene was also transformed into a NAD(P)H dehydrogenase (NDH1) deficient mutant of Synechocystis PCC 6803 (strain M55) (T. Ogawa, Proc. Natl. Acad. Sci. USA 88 (1991) 4275–4279). Phenotypic characterisation of the double mutant supported our conclusion that both the NAD(P)H dehydrogenase complex and FNR contribute independently to the quinone cytochrome b6f reductase step in PS I-dependent cyclic electron transfer. The distribution, binding properties and function of FNR in the model cyanobacterium Synechocystis PCC 6803 will be discussed

    Pathogenic NR2F1 variants cause a developmental ocular phenotype recapitulated in a mutant mouse model.

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    Pathogenic NR2F1 variants cause a rare autosomal dominant neurodevelopmental disorder referred to as the Bosch-Boonstra-Schaaf Optic Atrophy Syndrome. Although visual loss is a prominent feature seen in affected individuals, the molecular and cellular mechanisms contributing to visual impairment are still poorly characterized. We conducted a deep phenotyping study on a cohort of 22 individuals carrying pathogenic NR2F1 variants to document the neurodevelopmental and ophthalmological manifestations, in particular the structural and functional changes within the retina and the optic nerve, which have not been detailed previously. The visual impairment became apparent in early childhood with small and/or tilted hypoplastic optic nerves observed in 10 cases. High-resolution optical coherence tomography imaging confirmed significant loss of retinal ganglion cells with thinning of the ganglion cell layer, consistent with electrophysiological evidence of retinal ganglion cells dysfunction. Interestingly, for those individuals with available longitudinal ophthalmological data, there was no significant deterioration in visual function during the period of follow-up. Diffusion tensor imaging tractography studies showed defective connections and disorganization of the extracortical visual pathways. To further investigate how pathogenic NR2F1 variants impact on retinal and optic nerve development, we took advantage of an Nr2f1 mutant mouse disease model. Abnormal retinogenesis in early stages of development was observed in Nr2f1 mutant mice with decreased retinal ganglion cell density and disruption of retinal ganglion cell axonal guidance from the neural retina into the optic stalk, accounting for the development of optic nerve hypoplasia. The mutant mice showed significantly reduced visual acuity based on electrophysiological parameters with marked conduction delay and decreased amplitude of the recordings in the superficial layers of the visual cortex. The clinical observations in our study cohort, supported by the mouse data, suggest an early neurodevelopmental origin for the retinal and optic nerve head defects caused by NR2F1 pathogenic variants, resulting in congenital vision loss that seems to be non-progressive. We propose NR2F1 as a major gene that orchestrates early retinal and optic nerve head development, playing a key role in the maturation of the visual system

    Inhibition of Cell Division Suppresses Heterocyst Development in Anabaena sp. Strain PCC 7120

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    When the filamentous cyanobacterium Anabaena PCC 7120 is exposed to combined nitrogen starvation, 5 to 10% of the cells along each filament at semiregular intervals differentiate into heterocysts specialized in nitrogen fixation. Heterocysts are terminally differentiated cells in which the major cell division protein FtsZ is undetectable. In this report, we provide molecular evidence indicating that cell division is necessary for heterocyst development. FtsZ, which is translationally fused to the green fluorescent protein (GFP) as a reporter, is found to form a ring structure at the mid-cell position. SulA from Escherichia coli inhibits the GTPase activity of FtsZ in vitro and prevents the formation of FtsZ rings when expressed in Anabaena PCC 7120. The expression of sulA arrests cell division and suppresses heterocyst differentiation completely. The antibiotic aztreonam, which is targeted to the FtsI protein necessary for septum formation, has similar effects on both cell division and heterocyst differentiation, although in this case, the FtsZ ring is still formed. Therefore, heterocyst differentiation is coupled to cell division but independent of the formation of the FtsZ ring. Consistently, once the inhibitory pressure of cell division is removed, cell division should take place first before heterocyst differentiation resumes at a normal frequency. The arrest of cell division does not affect the accumulation of 2-oxoglutarate, which triggers heterocyst differentiation. Consistently, a nonmetabolizable analogue of 2-oxoglutarate does not rescue the failure of heterocyst differentiation when cell division is blocked. These results suggest that the control of heterocyst differentiation by cell division is independent of the 2-oxoglutarate signal

    Persistent photoconductivity in uniforndy and selectively silicon doped AlAs / GaAs short period superlattices

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    Hall and photo-Hall measurements have been carried out between 4 K et 400 K on MBE deposited AlAs / GaAs superiattices (SPS) with short period (25 Å << P<50P < 50 Å) SPSs were uniformly or selectively doped with silicon. Galvanomagnetic measurements show that SPSs exhibited an electrical behaviour similar to that of Alx_xGal1−x_{1-x}As : Si alloy (0.32<x<0.350.32 < x < 0.35). The Hall mobility was increased under illumination and persistent photoconductivity (PPC) was observed at low temperature (DX center). Ibermal annealing of PPC was performed by increasing the measurement temperature. Two plateaus are observed in the nH(T)n_{\rm H}(T) curves in uniformly doped SPSs whereas only one plateau was present in selectively doped SPSs. These experimental results are interpreted in terms of the multibarrier model of the DX center recently proposed in AIx_xGal1−x_{1-x}As: Si.Nous présentons des résultats de mesures d'effet Hall et photo-Hall obtenus entre 4 K et 400 K dans des superréseaux AlAs / GaAs de courtes périodes (25 Å << P<50P < 50 Å) déposées par MBE et dopées au silicium de manière uniforme ou sélectivement dans GaAs. Les mesures de concentration de porteurs et de mobilité par effet Hall à l'obscurité montrent que ce type de SPS (short period superiattice) présente un comportement électrique voisin de l'alliage AIx_xGal1−x_{1-x}As: Si de teneur en aluminium équivalente (0.32<x<0.350.32 < x < 0.35). Les mesures de photo-Hall à basse température montrent que ces SPS présentent également une photeconductivité persistente (PPC) et une augmentation de mobilité sous éclairement. La présence d'un plateau de PPC à basse temperature (T<90T < 90 K) est caractéristique du centre métastable DX dans tous les cas. Des mesures de décroissance du nombre de porteurs mesurés à l'obscurité aprés éclairement quand la température augmente (capture thermique), mettent en évidence la présence de deux plateaux correspondant à deux barrières thermiques de l'état métastable du centre DX dans les SPS à dopage uniforme, alors que les SPS dopées sélectivement dans GaAs ne présentent qu'une seule barrière (un seul plateau). Ainsi la microstructure GaAs / AlAs apparaît être un outil particulièrement intéressant pour confirmer la validité du modèle multibarrière proposé récemment pour le centre DX dans Alx_xGal1−x_{1-x} As: Si

    Iron Starvation Leads to Oxidative Stress in Anabaena sp. Strain PCC 7120

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    We establish here that iron deficiency causes oxidative stress in the cyanobacterium Anabaena sp. strain PCC 7120. Iron starvation leads to a significant increase in reactive oxygen species, whose effect can be abolished by treatment with the antioxidant tempol (4-hydroxy-2,2,6,6-tetramethylpiperidine 1-oxyl). Oxidative stress induced by iron starvation could be a common feature of photosynthetic bacteria
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