Structural analysis of the photosystem I supercomplex of cyanobacteria induced by iron deficiency

Here we describe the three-dimensional structure of the newly discovered CP43'-photosystem I (PSI) supercomplex of cyanobacteria calculated by single-particle analysis of images obtained by electron cryomicroscopy (cryo-EM). This large membrane protein complex has a molecular mass of approximately 2 MDa and is found in cyanobacteria when grown in iron deficient media. It is composed of a reaction center trimer surrounded by 18 subunits of the chlorophyll a binding CP43'protein, encoded by the isiA gene, which increases the light harvesting capacity of PSI by approximately 70%. By modeling higher-resolution structural data obtained from X-ray crystallography into the three-dimensional (3D) cryo-EM map, we have been able to gain a better understanding of the structure and functional properties of this supermolecular complex. We have identified three separate clusters of chlorophyll molecules at the periphery of the PSI core which may aid energy transfer from the CP43' antenna ring to the reaction center. Moreover, it is shown that despite the replacement of ferredoxin with flavodoxin as an electron acceptor under iron stress conditions, the 3D map has density to accommodate the extrinsic proteins, PsaC, PsaD, and PsaE. The presence of these three proteins was also confirmed by immunoblotting.     

Detecting plant spatial pattern change after disperser loss: A simulation and a case study

Disruption of seed dispersal processes may affect plant population spatial structure. We used a spatial simulation model and an empirical case study to assess the conditions under which the loss of seed dispersers has a detectable effect on a species' spatial pattern. Our simulation experiments suggested that detecting spatial change following disperser loss will be difficult, except when rates of fruit removal are initially high and then completely disappear. To contextualize the simulation modeling, we used spatial point pattern analyses to characterize the spatial pattern of two large-seeded species (Leucopogon nutans, a fire-killed seeder shrub and Macrozamia riedlei, a long-lived, resprouting cycad) in the jarrah (Eucalyptus marginata) forests of southwestern Australia. The plant species' primary disperser, the emu (Dromaius novaehollandiae), was absent from one of the sites we considered, but present at the other two. There was no detectable difference for either plant species in the strength of aggregation between sites with and without emu. However, even if disperser loss may not greatly affect local spatial structure for most plant species, it is likely to be important for long distance dispersal and genetic structuring of populations, so accurate characterization of the dispersal kernel is critical, especially in terms of plant emigration.     

Tree line identification from pollen data: beyond the limit?

Aim The boreal tree line is a prominent biogeographic feature, the position of which reflects climatic conditions. Pollen is the key sensor used to reconstruct past tree line patterns. Our aims in this study were to investigate pollen–vegetation relationships at the boreal tree line and to assess the success of a modified version of the biomization method that incorporates pollen productivity and dispersal in distinguishing the tree line. Location Northern Canada (307 sites) and Alaska (316 sites). Methods The REVEALS method for estimating regional vegetation composition from pollen data was simplified to provide correction factors to account for differential production and dispersal of pollen among taxa. The REVEALS‐based correction factors were used to adapt the biomization method and applied as a set of experiments to pollen data from lake sediments and moss polsters from the boreal tree line. Proportions of forest and tundra predicted from modern pollen samples along two longitudinal transects were compared with those derived from a vegetation map by: (1) a tally of ‘correct’ versus ‘incorrect’ assignments using vegetation in the relevant map pixels, and (2) a comparison of the shape and position of north–south forest‐cover curves generated from all transect pixels and from pollen data. Possible causes of bias in the misclassifications were assessed. Results Correcting for pollen productivity alone gave fewest misclassifications and the closest estimate of the modern mapped tree line position (Canada, + 300 km; Alaska, + 10 km). In Canada success rates were c. 40–70% and all experiments over‐predicted forest cover. Most corrections improved results over uncorrected biomization; using only lakes improved success rates to c. 80%. In Alaska success rates were 70–80% and classification errors were more evenly distributed; there was little improvement over uncorrected biomization. Main conclusions Corrected biomization should improve broad‐scale reconstructions of spatial patterns in forest/non‐forest vegetation mosaics and across climate‐sensitive ecotones. The Canadian example shows this is particularly the case in regions affected by taxa with extremely high pollen productivity (such as Pinus). Improved representation of actual vegetation distribution is most likely if pollen data from lake sediments are used because the REVEALS algorithm is based on the pollen dynamics of lake‐based systems.     

Fire-stimulated reproduction in the resprouting,non-serotinous conifer Podocarpus drouynianus (Podocarpaceae): the impact of a changing fire regime

Species with fire stimulated reproduction (fsr) are common in Mediterranean climate ecosystems. We investigated how season of, and time since, fire affects seed production in Podocarpus drouynianus F. Muell., a dioecious resprouting coniferous shrub endemic to the jarrah (Eucalyptus marginata Sm.) forests of southwestern Australia, and if the now largely managed fire regime in these forests poses a risk to its persistence. We hypothesised that, like other species showing fsr, seed production in P. drouynianus would be limited to the first few years following fire and seed set would be lower after spring burns. Mature plants regenerated rapidly from buried stem tissue (lignotuber) after fire, producing abundant sporophylls in autumn 12–18 months later. Stands burnt in autumn showed peak seed production 1 year later, while for those burned in spring, peak seed production was delayed until the second autumn after fire. Limited seed production occurred for up to 3 years following fire, but no seed production was observed in longer unburnt (>10 years since fire) stands. While we did not observe a significant impact of fire season on seed production, seed weight and viability were lower for spring-burnt plants. Population-level effects associated with plant density may also have negative impacts on P. drouynianus demography, with females within a small population burnt in autumn producing very few seeds 12 months following fire. Interactions between climate change, fire regimes and fire management practices need to be considered in order to best safeguard the long-term persistence of this conifer species.     

The composition and structure of photosystem I‐associated antenna from Cyanidioschyzon merolae

Red algae contain two types of light‐harvesting antenna systems, the phycobilisomes and chlorophyll a binding polypeptides (termed Lhcr), which expand the light‐harvesting capacity of the photosynthetic reaction centers. In this study, photosystem I (PSI) and its associated light‐harvesting proteins were isolated from the red alga Cyanidioschyzon merolae. The structural and functional properties of the largest PSI particles observed were investigated by biochemical characterization, mass spectrometry, fluorescence emission and excitation spectroscopy, and transmission electron microscopy. Our data provide strong evidence for a stable PSI complex in red algae that possesses two distinct types of functional peripheral light‐harvesting antenna complex, comprising both Lhcr and a PSI‐linked phycobilisome sub‐complex. We conclude that the PSI antennae system of red algae represents an evolutionary intermediate between the prokaryotic cyanobacteria and other eukaryotes, such as green algae and vascular plants.     

Measuring emotion in advertising research: prefrontal brain activity

With the current interest in the role of emotion in advertising and advertising research, there has been an increasing interest in the use of various brain activity measures to access nonverbal emotional responses. One such approach relies on measuring the difference between left and right hemisphere prefrontal cortical activity to assess like and dislike. This approach is based on electroencephalography (EEG) and neuroimaging work, suggesting that the approach/withdrawal (frequently but not always associated with like/dislike) dimension of emotion is indicated by the balance of activity between the left and right prefrontal cortex. Much of this work was initiated by Richard Davidson in the early 1990s. An early study by Davidson et al. measured brain electrical activity to assess patterns of activation during the experience of happiness and disgust. The authors reported that disgust was found to be associated with increased right-sided activation in the frontal and anterior temporal regions compared with happiness. In contrast, happiness was found to be accompanied by left-sided activation in the anterior temporal region compared with disgust. Early reports suggested that frontal laterality indexes motivational valence with positive emotions (happy, like) associated with left greater than the right frontal activity and vice versa. Although these findings appear to be consistent with personality traits (e.g., optimism pessimism), state changes in frontal laterality appears to index approach withdraw rather than emotional valence. Interestingly, the behavioral and motivational correlates of prefrontal asymmetric activity are not restricted to humans or even primates but have been observed in numerous species such as birds and fish (see [4]). Henceforth, we use the term motivational valence (MV) rather than the more cumbersome term approach withdraw.     

The 1.45 A three-dimensional structure of C-phycocyanin from the thermophilic cyanobacterium Synechococcus elongatus

The conversion of solar radiation to chemical energy by photosynthetic organisms provides the primary driving force for life on earth. Light energy is captured by a variety of pigments, usually bound to proteins, which vary with different types of organisms. We report here the 1.45 A resolution three-dimensional structure of one such pigment protein, C-phycocyanin, from Synechococcus elongatus. The structure is at the highest resolution achieved for any such phycobiliprotein. This level of resolution was made possible by implementing a novel crystallization method whereby nucleation is decoupled from subsequent growth, by incubating crystallizing drops for 7h under nucleation conditions and then transferring them to metastable conditions for growth. This is done without touching the crystallization drops throughout the process.     

New enzymes from environmental cassette arrays: functional attributes of a phosphotransferase and an RNA-methyltransferase

By targeting gene cassettes by polymerase chain reaction (PCR) directly from environmentally derived DNA, we are able to amplify entire open reading frames (ORFs) independently of prior sequence knowledge. Approximately 10% of the mobile genes recovered by these means can be attributed to known protein families. Here we describe the characterization of two ORFs which show moderate homology to known proteins: (1) an aminoglycoside phosphotransferase displaying 25% sequence identity with APH(7") from Streptomyces hygroscopicus, and (2) an RNA methyltransferase sharing 25%-28% identity with a group of recently defined bacterial RNA methyltransferases distinct from the SpoU enzyme family. Our novel genes were expressed as recombinant products and assayed for appropriate enzyme activity. The aminoglycoside phosphotransferase displayed ATPase activity, consistent with the presence of characteristic Mg(2+)-binding residues. Unlike related APH(4) or APH(7") enzymes, however, this activity was not enhanced by hygromycin B or kanamycin, suggesting the normal substrate to be a different aminoglycoside. The RNA methyltransferase contains sequence motifs of the RNA methyltransferase superfamily, and our recombinant version showed methyltransferase activity with RNA. Our data confirm that gene cassettes present in the environment encode folded enzymes with novel sequence variation and demonstrable catalytic activity. Our PCR approach (cassette PCR) may be used to identify a diverse range of ORFs from any environmental sample, as well as to directly access the gene pool found in mobile gene cassettes commonly associated with integrons. This gene pool can be accessed from both cultured and uncultured microbial samples as a source of new enzymes and proteins.     

Metabolism of N tau-methylhistidine by mice.

Mice and rats were injected with tracer doses of radioactive N tau-[Me-14C]methylhistidine in order to determine the recovery of the injected radioactivity and the extent of the metabolism of N tau-methylhistidine. In the first 27 h after injection, 96.3, 78.0 and 97.5% of radioactivity was excreted by female mice, male mice and male rats respectively. Recovery after 5 days of collection was 98.4 and 92.8% for female and male mice respectively. However, radioactivity associated with N tau-methylhistidine or its acetylated derivative accounted for 44, 86.5 and 96.0% of the excreted radioactivity for female mice, male mice and rats respectively. In female mice the remaining excreted radioactivity was associated with four major peaks of activity when the metabolites were separated by cation-exchange chromatography. In male mice there were only three of these metabolites present. After chromatographic purification, one metabolite was identified by mass spectroscopy to be 1-methylimidazole-4-acetic acid. Examination of the possible sources of this metabolite indicates that, in mice, N tau-methylhistidine is decarboxylated and enters the chain of reactions common to histamine metabolism. Such extensive metabolism precludes the use of N tau-methylhistidine excretion as an index of myofibrillar protein breakdown in mice.