Protoplasts from barley (Hordeum vulgare), pea (fisum sativum), wheat (Trificum aesfivum), and spinach (Spinacia oleracea) leaves were fradionated into chloroplast-and mitochondrionenriched fractions. Pyruvate dehydrogenase complex capacities in mitochondria (mtPDC) and chloroplasts (cpPDC) were measured in appropriate fractions under conditions optimal for each isozyme. The total cellular capacity of PDC was similar in barley and pea but about 50% lower in wheat and spinach. In pea a distribution of 87% mtPDC and 13% cpPDC was found on a cellular basis. In barley, wheat, and spinach the subcellular distribution was the opposite, with about 15% mtPDC and 85% cpPDC. cpPDC adivity was constant at about 0
Mitochondria fulfill important functions in photosynthetic cells not only in darkness but also in light. Mitochondrial oxidative phosphorylation is probably the main mechanism to supply ATP for extrachloroplastic functions in both conditions. Furthermore, during photosynthesis mitochondrial electron transport is important for regulation of the redox balance in the cell. This makes mitochondrial function an integral part of a flexible metabolic system in the photosynthetic cell. This flexibility is probably very important in order to allow the metabolism to override disturbances caused by the changing environment which plants are adapted to.
Photosynthesis was characterized for the unicellular green alga Coccomyxa sp., grown at low inorganic carbon (Ci) concentrations, and compared with Chlamydomonas reinhardtii, which had been grown so that the CO2 concentrating mechanism (CCM) was expressed, and with protoplasts isolated from the C3 plant barley (Hordeum vulgare). Chlamydomonas had a significantly higher Ci‐use efficiency of photosynthesis, with an initial slope of the Ci‐response curve of 0.7 mol(gChl)−1 h−1 mmol Cim−3)−1, as compared to 0.3 and 0.23 mol(gChl)−1 h−1 (mmol Cim−3)−1 for Coccomyxa and barley, respectively. The affinity for Ci was also higher in Chlamydomonas, as the half maximum rate of photosynthesis [K0.5 (Ci)] was reached at 0.18 mol m−3, as compared to 0.30 and 0.45 mol m−3 for Coccomyxa and barley, respectively. Ethoxyzolamide (EZ), an inhibitor of the enzyme carbonic anhydrase (CA) and the CCM, caused a 17‐fold decrease in the initial slope of the photosynthetic Cj‐response curve in Chlamydomonas, but only a 1.5‐ to two‐fold decrease in Coccomyxa and barley. The photosynthetic light‐response curve showed further similarities between barley and Coccomyxa. The rate of bending of the curve, described by the convexity parameter, was 0.99 (sharp bending) and 0.81–0.83 (gradual bending) for cells grown under low and high light, respectively. In contrast, the maximum convexity of Chlamydomonas was 0.85. The intrinsically lower convexity of Chlamydomonas is suggested to result from the diversion of electron transport from carbon fixation to the CCM. Taken together, these results suggest that Coccomyxa does not possess a CCM and due to this apparent lack of a CCM, we propose that Coccomyxa is a better cell model system for studying C3 plant photosynthesis than many algae currently used.
The Ca2+-stimulated type 1 adenylyl cyclase (AC1) contributes to several forms of synaptic plasticity and is the only known neurospecific adenylyl cyclase. Furthermore, the protein and mRNA levels of AC1 undergo a circadian oscillation in the pineal gland, and AC1 may play a pivotal role in regulating nocturnal melatonin synthesis. To better understand the expression of AC1, we isolated mouse genomic DNA clones of AC1. The transcription and translation start regions of mouse AC1 share extensive homologies with the bovine counterpart. The upstream proximal region has potential binding sites for transcription factors, including the steroid receptor family, the E-box factors, and Sp1. A 280-bp fragment that contains the transcription start site directed reporter gene expression in cultured cortical neurons and pinealocytes functioning as a basal neuro- and pineal-directed promoter. Interestingly, pinealocyte expression of the reporter gene was inhibited by increases in cAMP. This cAMP sensitivity may explain why AC1 mRNA in the pineal is low at night when cAMP is elevated and high during the day when cAMP signals drop. An adjacent 330-bp fragment interacted specifically with nuclear factor(s) that we designate binary E-box factor (BEF). Methylation interference and DNase I footprinting identified the BEF-binding site sequence as 5'-CCAAGGTCACGTGGC-3'. When linked to the basal tissue-directed promoter, this 15-bp sequence further enhanced reporter expression in neurons and pinealocytes. We propose that this 15-bp sequence may contribute to increased expression of AC1 in neurons and pinealocytes relative to other cells.
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