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Krause, D.

Publications and source records attributed to Krause, D..

6 recordsLinked to original sources

Analyses of Genome-Wide Recombination in Sulfolobus islandicus

Recombination is a fundamental process that both maintains genome integrity and dramatically shapes evolutionary dynamics in populations from all domains of life. Here we cross two genetically distinct but highly syntenic sympatric strains of Sulfolobus (Saccharolobus) islandicus and identify the pattern of recombination across the archaeal chromosome. We quantify recombination breakpoints occurring at neutral chromosomal markers through high-throughput short read Illumina sequencing of selected recombinant strains. We focus on three datasets that provide different, but corroborating information: i) individual colony sequences, ii) large colony pools of strains sorted by background genotype, and iii) a large colony pool of combined background genotypes. For each dataset we apply a newly designed and optimized computational tool called MARBL (Multiply Anchored Recombination Breakpoint Locator) for quantifying breakpoints and testing their statistical significance relative to sources of error introduced by sequencing technology. By combining the results of each dataset, we determine the short tract recombination preference in this organism and discuss its implications for mechanisms of recombination in archaea. We demonstrate uniform rates of recombination throughout the chromosome, which differ from previous studies of natural variation and discuss this in light of previous data suggesting speciation occurs through selection against inter-species hybrids. Importance StatementRecombination is a critical process in all domains of life that facilitates the essential repair of chromosomes and exchanges variation between organisms. Here we investigate genome-wide patterns of recombination in the model archaeaon Saccharolobus islandicus to better understand mechanistic aspects of their DNA exchange and recombination processes. Our observations of recombination in a controlled laboratory setting identified patterns of recombination that support the ability to incorporate short pieces of DNA in a piecemeal fashion from larger transferred segments. By comparing these observations to inferred recombination from natural populations, we can gain insight into evolutionary parameters including natural selection that would be otherwise difficult to determine from genomic evidence alone.

microbiology↗

Single-shot impulsive stimulated Brillouin microscopy by tailored ultrashort pulses

Brillouin microscopy has become an important tool for investigating the mechanical properties of tissue. The recently developed Impulsive stimulated Brillouin Scattering (ISBS) promises a label-free, non-invasive measurements of viscoelastic properties of transparent samples and offers the potential for a high temporal resolution. However, the spatial resolution of ISBS is currently limited, which hinders its transfer to real-world applications. Increasing the spatial resolution of ISBS leads to an increase in the energy density of the pump beams, which requires a balancing of the excitation parameters to stay below the phototoxic threshold. This paper focuses on the influences of different excitation parameters on the spatial, temporal and spectral resolution and their optimal values. Combined with the adoption of a noise suppressing window function, a measurement rate of 20{micro}s/pixel in hydrogel is achieved, which is promising for fast 3D imaging. The presented advanced impulsive stimulated Brillouin microscopy can be applied for fast tissue elastography to-ward disease studies.

biophysics↗

Lipidome visualisation, comparison, and analysis in a vector space

A shallow neural network was used to embed lipid structures in a 2- or 3-dimensional space with the goal that structurally similar species have similar vectors. Tests on complete lipid databanks show that the method automatically produces distributions which follow conventional lipid classifications. The embedding is accompanied by the web-based software, Lipidome Projector. This displays user lipidomes as 2D or 3D scatterplots for quick exploratory analysis, quantitative comparison and interpretation at a structural level. Author summaryLipids are not just the basis of membranes. They carry signals and metabolic energy. This means that the presence, absence, and quantity of lipids reflects a cells biochemical state - starving, nourished, sick or healthy. Lipidomics (measuring all lipids in a biological specimen) provides lists of the chemical species and their quantities. We have used a shallow neural network from natural language modelling to embed lipids in a continuous vector space. Firstly, this means that similar molecules have similar positions in this space. Conventional lipid categories cluster automatically. Secondly, the accompanying web-based software, Lipidome Projector imports a lipidome and displays it as a set of points. Reading several lipidomes at once allows quantitative and structural comparisons. Combined with the ability to show structure and abundance diagrams, the software allows exploratory analysis and interpretation of lipidomics datasets.

bioinformatics↗

The capsular polysaccharide obstructs wall teichoic acid functions in Staphylococcus aureus

BackgroundThe cell envelope of Staphylococcus aureus contains two major secondary cell wall glycopolymers: capsular polysaccharide (CP) and wall teichoic acid (WTA). Both the CP and the WTA are attached to the cell wall and play distinct roles in S. aureus colonization, pathogenesis, and bacterial evasion of host immune defenses. ObjectiveWe aimed to investigate whether CP interferes with WTA-mediated properties. MethodsStrains with natural heterogeneous expression of CP, strains with homogeneous high CP expression and CP-deficient strains were compared to WTA deficient controls regarding WTA dependent phage binding, cell adhesion, IgG deposition, and virulence in vivo. ResultsWTA-mediated phage adsorption, specific antibody deposition and cell adhesion were negatively correlated with CP expression. WTA, but not CP, enhanced the bacterial burden in a mouse abscess model, while CP overexpression resulted in intermediate virulence in vivo. ConclusionsCP protects the bacteria from WTA-dependent opsonization and phage binding. This protection comes at the cost of diminished adhesion to host cells. The highly complex regulation and mostly heterogeneous expression of CP has probably evolved to ensure the survival and optimal physiological adaptation of the bacterial population as a whole.

microbiology↗

DynaTMT: A user-friendly tool to process combined SILAC/TMT data

The measurement of protein dynamics by proteomics to study cell remodeling has seen increased attention over the last years. This development is largely driven by a number of technological advances in proteomics methods. Pulsed stable isotope labeling in cell culture (SILAC) combined with tandem mass tag (TMT) labeling has evolved as a gold standard for profiling protein synthesis and degradation. While the experimental setup is similar to typical proteomics experiments, the data analysis proves more difficult: After peptide identification through search engines, data extraction requires either custom scripted pipelines or tedious manual table manipulations to extract the TMT-labeled heavy and light peaks of interest. To overcome this limitation, which deters researchers from using protein dynamic proteomics, we developed a user-friendly, browser-based application that allows easy and reproducible data analysis without the need for scripting experience. In addition, we provide a python package that can be implemented in established data analysis pipelines. We anticipate that this tool will ease data analysis and spark further research aimed at monitoring protein translation and degradation by proteomics.

bioinformatics↗

LAMP3 is critical for surfactant homeostasis in mice

Lysosome-associated membrane glycoprotein 3 (LAMP3) is a type I transmembrane protein of the LAMP protein family with a cell-type-specific expression in alveolar type II cells in mice and hitherto unknown function. In type II pneumocytes, LAMP3 is localized in lamellar bodies, secretory organelles releasing pulmonary surfactant into the extracellular space to lower surface tension at the air/liquid interface. The physiological function of LAMP3, however, remains enigmatic. We generated Lamp3 knockout mice by CRISPR/Cas9. LAMP3 deficient mice are viable with an average life span and display regular lung function under basal conditions. The levels of a major hydrophobic protein component of pulmonary surfactant, SP-C, are strongly increased in the lung of Lamp3 knockout mice, and the lipid composition of the bronchoalveolar lavage shows mild but significant changes, resulting in alterations in surfactant functionality. In ovalbumin-induced experimental allergic asthma, the changes in lipid composition are aggravated, and LAMP3-deficient mice exert an increased airway resistance. Our data suggest a critical role of LAMP3 in the regulation of pulmonary surfactant homeostasis and normal lung function.

biochemistry↗