Photochemical characterization of a novel fungal rhodopsin from Phaeosphaeria nodorum

Eukaryotic microbial rhodopsins are widespread bacteriorhodopsin-like proteins found in many lower eukaryotic groups including fungi. Many fungi contain multiple rhodopsins, some significantly diverged from the original bacteriorhodopsin template. Although few fungal rhodopsins have been studied biophysically, both fast-cycling light-driven proton pumps and slow-cycling photosensors have been found. The purpose of this study was to characterize photochemically a new subgroup of fungal rhodopsins, the so-called auxiliary group. The study used the two known rhodopsin genes from the fungal wheat pathogen, Phaeosphaeria nodorum. One of the genes is a member of the auxiliary group while the other is highly similar to previously characterized proton-pumping Leptosphaeria rhodopsin. Auxiliary rhodopsin genes from a range of species form a distinct group with a unique primary structure and are located in carotenoid biosynthesis gene cluster. Amino acid conservation pattern suggests that auxiliary rhodopsins retain the transmembrane core of bacteriorhodopsins, including all residues important for proton transport, but have unique polar intramembrane residues. Spectroscopic characterization of the two yeast-expressed Phaeosphaeria rhodopsins showed many similarities: absorption spectra, conformation of the retinal chromophore, fast photocycling, and carboxylic acid protonation changes. It is likely that both Phaeosphaeria rhodopsins are proton-pumping, at least in vitro.We suggest that auxiliary rhodopsins have separated from their ancestors fairly recently and have acquired the ability to interact with as yet unidentified transducers, performing a photosensory function without changing their spectral properties and basic photochemistry

Light Controls Growth and Development via a Conserved Pathway in the Fungal Kingdom

Light inhibits mating and haploid fruiting of the human fungal pathogen Cryptococcus neoformans, but the mechanisms involved were unknown. Two genes controlling light responses were discovered through candidate gene and insertional mutagenesis approaches. Deletion of candidate genes encoding a predicted opsin or phytochrome had no effect on mating, while strains mutated in the white collar 1 homolog gene BWC1 mated equally well in the light or the dark. The predicted Bwc1 protein shares identity with Neurospora crassa WC-1, but lacks the zinc finger DNA binding domain. BWC1 regulates cell fusion and repression of hyphal development after fusion in response to blue light. In addition, bwc1 mutant strains are hypersensitive to ultraviolet light. To identify other components required for responses to light, a novel self-fertile haploid strain was created and subjected to Agrobacterium-mediated insertional mutagenesis. One UV-sensitive mutant that filaments equally well in the light and the dark was identified and found to have an insertion in the BWC2 gene, whose product is structurally similar to N. crassa WC-2. The C. neoformans Bwc1 and Bwc2 proteins interact in the yeast two-hybrid assay. Deletion of BWC1 or BWC2 reduces the virulence of C. neoformans in a murine model of infection; the Bwc1-Bwc2 system thus represents a novel protein complex that influences both development and virulence in a pathogenic fungus. These results demonstrate that a role for blue/UV light in controlling development is an ancient process that predates the divergence of the fungi into the ascomycete and basidiomycete phyla

Photoreactions and Structural Changes of Anabaena Sensory Rhodopsin

Anabaena sensory rhodopsin (ASR) is an archaeal-type rhodopsin found in eubacteria. The gene encoding ASR forms a single operon with ASRT (ASR transducer) which is a 14 kDa soluble protein, suggesting that ASR functions as a photochromic sensor by activating the soluble transducer. This article reviews the detailed photoreaction processes of ASR, which were studied by low-temperature Fourier-transform infrared (FTIR) and UV-visible spectroscopy. The former research reveals that the retinal isomerization is similar to bacteriorhodopsin (BR), but the hydrogen-bonding network around the Schiff base and cytoplasmic region is different. The latter study shows the stable photoproduct of the all-trans form is 100% 13-cis, and that of the 13-cis form is 100% all-trans. These results suggest that the structural changes of ASR in the cytoplasmic domain play important roles in the activation of the transducer protein, and photochromic reaction is optimized for its sensor function

Light regulation of metabolic pathways in fungi

Light represents a major carrier of information in nature. The molecular machineries translating its electromagnetic energy (photons) into the chemical language of cells transmit vital signals for adjustment of virtually every living organism to its habitat. Fungi react to illumination in various ways, and we found that they initiate considerable adaptations in their metabolic pathways upon growth in light or after perception of a light pulse. Alterations in response to light have predominantly been observed in carotenoid metabolism, polysaccharide and carbohydrate metabolism, fatty acid metabolism, nucleotide and nucleoside metabolism, and in regulation of production of secondary metabolites. Transcription of genes is initiated within minutes, abundance and activity of metabolic enzymes are adjusted, and subsequently, levels of metabolites are altered to cope with the harmful effects of light or to prepare for reproduction, which is dependent on light in many cases. This review aims to give an overview on metabolic pathways impacted by light and to illustrate the physiological significance of light for fungi. We provide a basis for assessment whether a given metabolic pathway might be subject to regulation by light and how these properties can be exploited for improvement of biotechnological processes

De novo Assembly of a 40 Mb Eukaryotic Genome from Short Sequence Reads: Sordaria macrospora, a Model Organism for Fungal Morphogenesis

Filamentous fungi are of great importance in ecology, agriculture, medicine, and biotechnology. Thus, it is not surprising that genomes for more than 100 filamentous fungi have been sequenced, most of them by Sanger sequencing. While next-generation sequencing techniques have revolutionized genome resequencing, e.g. for strain comparisons, genetic mapping, or transcriptome and ChIP analyses, de novo assembly of eukaryotic genomes still presents significant hurdles, because of their large size and stretches of repetitive sequences. Filamentous fungi contain few repetitive regions in their 30–90 Mb genomes and thus are suitable candidates to test de novo genome assembly from short sequence reads. Here, we present a high-quality draft sequence of the Sordaria macrospora genome that was obtained by a combination of Illumina/Solexa and Roche/454 sequencing. Paired-end Solexa sequencing of genomic DNA to 85-fold coverage and an additional 10-fold coverage by single-end 454 sequencing resulted in ∼4 Gb of DNA sequence. Reads were assembled to a 40 Mb draft version (N50 of 117 kb) with the Velvet assembler. Comparative analysis with Neurospora genomes increased the N50 to 498 kb. The S. macrospora genome contains even fewer repeat regions than its closest sequenced relative, Neurospora crassa. Comparison with genomes of other fungi showed that S. macrospora, a model organism for morphogenesis and meiosis, harbors duplications of several genes involved in self/nonself-recognition. Furthermore, S. macrospora contains more polyketide biosynthesis genes than N. crassa. Phylogenetic analyses suggest that some of these genes may have been acquired by horizontal gene transfer from a distantly related ascomycete group. Our study shows that, for typical filamentous fungi, de novo assembly of genomes from short sequence reads alone is feasible, that a mixture of Solexa and 454 sequencing substantially improves the assembly, and that the resulting data can be used for comparative studies to address basic questions of fungal biology

Emerging futures for Sao Tome and Principe's International Airport Campus

CIVINS (Civilian Institutions) Thesis documentSao Tome and Principe has recently reached the completion point for the United Nation's Highly Indebted and Poor Countries (HIPC) program and is now looking to develop a sustainable and prosperous economy (International Monetary Fund 2007). However, the existing international airport is failing to meet the safety, security and capacity requirements to further the country's economic development. Construction of an airport security fence is underway that will cutoff several thousand residents from the water sources and will redirect routes to the markets of Sao Tome. This Master's project utilizes scenario planning to explore possible futures for the expansion of Sao Tome and Principe's international airport. The scenario development process is intended to facilitate dialogue within the country about the future by which policy and development decisions can be made. A wide range of topics such as environment, history, culture, economy, social structure, land use, domestic policy and infrastructure were studied in order to make educated projections. potential benefits, dangers, and conflicts between the issues were considered in order to design planning scenarios that present both positive and negative futures. The result was four primary scenarios for the international airport and surrounding campus that explore opportunities and externalities inherent in such a project with broad and drastic impacts. The representative scenarios included in this final report demonstrate a broad range of alternative futures such as: dramatic expansion, no expansion, or conservative expansion of the airport. Each of the scenarios is presented through various means to describe the possible future including: written analysis, story telling (narratives), sketches, mapping, and photo vignettes. This report is intended to stimulate discussion regarding the future Sao Tome and Principe.http://archive.org/details/emergingfuturesf109452991Funding number: N62271-97-G-0056.Approved for public release; distribution is unlimited

Religious motifs in the songs performed by football fans

Tekst ukazuje, jak kibice piłki nożnej używają religijnych motywów w swoich przyśpiewkach. Istnieje kilka przyczyn tego rodzaju działania. Pieśni religijne stanowią istotny element polskiej kultury. Parafrazowanie ich ułatwia zapamiętanie rytmu. Klub piłkarski bardzo często bywa określany „świętym”. Kibice porównują klub do Boga, co dodaje wagi i wprowadza klub w sferę sacrum. Tego rodzaju przyśpiewki pokazują również, że klub piłkarski jest uważany za święty przez kibiców tworzących tego typu przyśpiewki.Text shows how football fans use religious motifs in their songs. There are a few reasons for that. Religious songs are an important part of Polish culture. Paraphrasing them makes it easier to remember the rhythm. Club is very often said to be “saint”. It is compared to the God. It adds importance and brings the club into sacrum zone. It also shows that club is regarded by sacrum by fans who create this kind of songs

“Devilish sympathetic, angels like hell” - about religious motifs on football supporters’ flags

In my paper I describe how Polish football fans use religious motifs on their flags. The reasons for it can be different. They compare their football club to God. They want to make opposite team’s supporters frightened (then they very often use devil motif) or they want to make their communicate more attractive. All of these with examples are described in the paper

Esters of 2-azidocarboxylic acid as starting materials for the synthesis of symmetrical pyrroles substituted heterocyclic systems

Poniższa praca skupia się na próbie otrzymania symetrycznego 2,5-dikarboksylowego pirolu podstawionego w pozycji 3 oraz 4 układami heterocyklicznymi. Substratami wyjściowymi do powyższych związków są 2-azydoestry, które najłatwiej można otrzymać z 2-bromoestrów. W trakcie prowadzonych badań zaproponowano szereg alternatywnych ścieżek wychodząc z 2-formylotiofenu oraz 2-formylopirydyny. Jednak z powodu braku czasu dalsze syntezy prowadzono tylko na pochodnych tiofenu. W rezultacie okazało się, iż największym problemem jest otrzymanie odpowiedniego 2-bromoestru, który stałby się substratem wyjściowym do otrzymania końcowego produktu.The following work is focusing on the attempt to obtain a symmetrical 2,5-dicarboxylic pyrrole substituted in position 3 and 4 of heterocyclic systems. The starting materials for the above compounds are 2-azidoesters which most easily can be prepared from 2-bromoesters. The during of research proposed a number of alternative paths starting from 2-thiophenecarboxaldehyde and 2-pyridinecarboxaldehyde. However, lack of time further syntheses were carried out only thiophene derivatives. As a result it turned out that receiving appropriate 2-bromoesters which would become initial substrate was a biggest problem for receiving the final produc