ALFA: First Operational Experience of the MPE/MPIA Laser Guide Star System for Adaptive Optics

The sodium laser guide star adaptive optics system ALFA has been constructed at the Calar Alto 3.5-m telescope. Following the first detection of the laser beacon on the wavefront sensor in 1997 the system is now being optimized for best performance. In this contribution we discuss the current status of the launch beam and the planned improvements and upgrades. We report on the performance level achieved when it is used with the adaptive optics system, and relate various aspects of our experience during operation of the system. We have begun to produce scientific results and mention two of these.Comment: 9 pages, 6 figures, LaTeX (spie.sty). SPIE conf proc 3353, Adaptive Optical System Technologies, March 199

On insertion-deletion systems over relational words

We introduce a new notion of a relational word as a finite totally ordered set of positions endowed with three binary relations that describe which positions are labeled by equal data, by unequal data and those having an undefined relation between their labels. We define the operations of insertion and deletion on relational words generalizing corresponding operations on strings. We prove that the transitive and reflexive closure of these operations has a decidable membership problem for the case of short insertion-deletion rules (of size two/three and three/two). At the same time, we show that in the general case such systems can produce a coding of any recursively enumerable language leading to undecidabilty of reachability questions.Comment: 24 pages, 8 figure

Cell-Free Synthesis of the Mitochondrial ADP/ATP Carrier Protein of Neurospora crassa

ADP/ATP carrier protein was synthesized in heterologous cell-free systems programmed with Neurospora poly(A)-containing RNA and homologous cell-free systems from Neurospora. The apparent molecular weight of the product obtained in vitro was the same as that of the authentic mitochondrial protein. The primary translation product obtained in reticulocyte lysates starts with formylmethionine when formylated initiator methionyl-tRNA (fMet-tRNAfMet) was present. The product synthesized in vitro was released from the ribosomes into the postribosomal supernatant. The evidence presented indicates that the ADP/ATP carrier is synthesized as a polypeptide with the same molecular weight as the mature monomeric protein and does not carry an additional sequence

Genome-wide transcription start site mapping of Bradyrhizobium japonicum grown free-living or in symbiosis – a rich resource to identify new transcripts, proteins and to study gene regulation

Background: Differential RNA-sequencing (dRNA-seq) is indispensable for determination of primary transcriptomes. However, using dRNA-seq data to map transcriptional start sites (TSSs) and promoters genome-wide is a bioinformatics challenge. We performed dRNA-seq of Bradyrhizobium japonicum USDA 110, the nitrogen-fixing symbiont of soybean, and developed algorithms to map TSSs and promoters. Results: A specialized machine learning procedure for TSS recognition allowed us to map 15,923 TSSs: 14,360 in free-living bacteria, 4329 in symbiosis with soybean and 2766 in both conditions. Further, we provide proteomic evidence for 4090 proteins, among them 107 proteins corresponding to new genes and 178 proteins with N-termini different from the existing annotation (72 and 109 of them with TSS support, respectively). Guided by proteomics evidence, previously identified TSSs and TSSs experimentally validated here, we assign a score threshold to flag 14 % of the mapped TSSs as a class of lower confidence. However, this class of lower confidence contains valid TSSs of low-abundant transcripts. Moreover, we developed a de novo algorithm to identify promoter motifs upstream of mapped TSSs, which is publicly available, and found motifs mainly used in symbiosis (similar to RpoN-dependent promoters) or under both conditions (similar to RpoD-dependent promoters). Mapped TSSs and putative promoters, proteomic evidence and updated gene annotation were combined into an annotation file. Conclusions: The genome-wide TSS and promoter maps along with the extended genome annotation of B. japonicum represent a valuable resource for future systems biology studies and for detailed analyses of individual non-coding transcripts and ORFs. Our data will also provide new insights into bacterial gene regulation during the agriculturally important symbiosis between rhizobia and legumes

Different Transport Pathways of Individual Precursor Proteins in Mitochondria

Transport of mitochondrial precursor proteins into mitochondria of Neurospora crassa was studied in a cellfree reconstituted system. Precursors were synthesized in a reticulocyte lysate programmed with Neurospora mRNA and transported into isolated mitochondria in the absence of protein synthesis. Uptake of the following precursors was investigated: apocytochrome c, ADP/ATP carrier and subunit 9 of the oligomycin-sensitive ATPase. Addition of high concentrations of unlabelled chemically prepared apocytochrome c (1–10 μM) inhibited the appearance in the mitochondrial of labelled cytochrome c synthesized in vitro because the unlabelled protein dilutes the labelled one and because the translocation system has a limited capacity [apparent V is 1–3 pmol × min−1× (mg mitochondrial protein)−1]. Concentrations of added apocytochrome c exceeding the concentrations of precursor proteins synthesized in vitro by a factor of about 104 did not inhibit the transfer of ADP/ATP carrier or ATPase subunit 9 into mitochondria. Carbonylcyanide m-chlorophenylhydrazone, an uncoupler of oxidative phosphorylation, inhibited transfer in vitro of ADP/ATP carrier and of ATPase subunit 9, but not of cytochrome c. These findings suggest that cytochrome c and the other two proteins have different import pathways into mitochondria. It can be inferred from the data presented that different 'receptors' on the mitochondrial surface mediate the specific recognition of precursor proteins by mitochondria as a first step in the transport process

Evaluating Oceanic Uptake of Atmospheric CCl4: A Combined Analysis of Model Simulations and Observations

We provide new estimates of the air‐sea flux of CCl4 using simulations from a global ocean biogeochemistry model (NEMO‐PlankTOM) in combination with depth‐resolved CCl4 observations from global oceanic databases. Estimates of global oceanic CCl4 uptake are derived from a range of model analyses, including prescribed parameterizations using reported values on hydrolysis and degradation, and analyses optimized using the global observational databases. We evaluate the sensitivity of our results to uncertainties in air‐sea gas exchange parameterization, estimation period, and circulation processes. Our best constrained estimate of ocean CCl4 uptake for the period 1996–2000 is 20.1 Gg/year (range 16.6–22.7), corresponding to estimates of the partial atmospheric lifetime with respect to ocean uptake of 124 (110–150) years. This new oceanic lifetime implies higher emissions of CCl4 than currently estimated and therefore a larger missing atmospheric source of CCl4

Response of the solar atmosphere to magnetic field evolution in a coronal hole region

Methods. We study an equatorial CH observed simultaneously by HINODE and STEREO on July 27, 2007. The HINODE/SP maps are adopted to derive the physical parameters of the photosphere and to research the magnetic field evolution and distribution. The G band and Ca II H images with high tempo-spatial resolution from HINODE/BFI and the multi-wavelength data from STEREO/EUVI are utilized to study the corresponding atmospheric response of different overlying layers. Results. We explore an emerging dipole locating at the CH boundary. Mini-scale arch filaments (AFs) accompanying the emerging dipole were observed with the Ca II H line. During the separation of the dipolar footpoints, three AFs appeared and expanded in turn. The first AF divided into two segments in its late stage, while the second and third AFs erupted in their late stages. The lifetimes of these three AFs are 4, 6, 10 minutes, and the two intervals between the three divisions or eruptions are 18 and 12 minutes, respectively. We display an example of mixed-polarity flux emergence of IN fields within the CH and present the corresponding chromospheric response. With the increase of the integrated magnetic flux, the brightness of the Ca II H images exhibits an increasing trend. We also study magnetic flux cancellations of NT fields locating at the CH boundary and present the obvious chromospheric and coronal response. We notice that the brighter regions seen in the 171 A images are relevant to the interacting magnetic elements. By examining the magnetic NT and IN elements and the response of different atmospheric layers, we obtain good positive linear correlations between the NT magnetic flux densities and the brightness of both G band (correlation coefficient 0.85) and Ca II H (correlation coefficient 0.58).Comment: 9 pages, 9 figures. A&A, in pres

Transport of Proteins into Mitochondria

The mitochondrial ADP/ATP carrier is an integral transmembrane protein of the inner membrane. It is synthesized on cytoplasmic ribosomes. Kinetic data suggested that this protein is transferred into mitochondria in a posttranslational manner. The following results provide further evidence for such a mechanism and provide information on its details. 1. In homologous and heterologous translation systems the newly synthesized ADP/ATP carrier protein is present in the postribosomal supernatant. 2. Analysis by density gradient centrifugation and gel filtration shows, that the ADP/ATP carrier molecules in the postribosomal fraction are present as soluble complexes with apparent molecular weights of about 120000 and 500000 or larger. The carrier binds detergents such as Triton X-100 and deoxycholate forming mixed micelles with molecular weights of about 200000–400000. 3. Incubation of a postribosomal supernatant of a reticulocyte lysate containing newly synthesized ADP/ATP carrier with mitochondria isolated from Neurospora spheroplasts results in efficient transfer of the carrier into mitochondria. About 20–30% of the transferred carrier are resistant to proteinase in whole mitochondria. The authentic mature protein is also largely resistant to proteinase in whole mitochondria and sensitive after lysis of mitochondria with detergent. Integrity of mitochondria is a prerequisite for translocation into proteinase resistant position. 4. The transfer in vitro into a proteinase-resistant form is inhibited by the uncoupler carbonyl-cyanide m-chlorophenylhydrazone but not the proteinase-sensitive binding. These observations suggest that the posttranslational transfer of ADP/ATP carrier occurs via the cytosolic space through a soluble oligomeric precursor form. This precursor is taken up by intact mitochondria into an integral position in the membrane. These findings are considered to be of general importance for the intracellular transfer of insoluble membrane proteins. They support the view that such proteins can exist in a water-soluble form its precursors and upon integration into the membrane undergo a conformational change. Uptake into the membrane may involve the cleavage of an additional sequence in some proteins, but this appears not to be a prerequisite as demonstrated by the ADP/ATP carrier protein