The Candida albicans Ku70 Modulates Telomere Length and Structure by Regulating Both Telomerase and Recombination

The heterodimeric Ku complex has been shown to participate in DNA repair and telomere regulation in a variety of organisms. Here we report a detailed characterization of the function of Ku70 in the diploid fungal pathogen Candida albicans. Both ku70 heterozygous and homozygous deletion mutants have a wild-type colony and cellular morphology, and are not sensitive to MMS or UV light. Interestingly, we observed complex effects of KU70 gene dosage on telomere lengths, with the KU70/ku70 heterozygotes exhibiting slightly shorter telomeres, and the ku70 null strain exhibiting long and heterogeneous telomeres. Analysis of combination mutants suggests that the telomere elongation in the ku70 null mutant is due mostly to unregulated telomerase action. In addition, elevated levels of extrachromosomal telomeric circles were detected in the null mutant, consistent with activation of aberrant telomeric recombination. Altogether, our observations point to multiple mechanisms of the Ku complex in telomerase regulation and telomere protection in C. albicans, and reveal interesting similarities and differences in the mechanisms of the Ku complex in disparate systems

First Complete Mitochondrial Genome Sequence from a Box Jellyfish Reveals a Highly Fragmented Linear Architecture and Insights into Telomere Evolution

Animal mitochondrial DNAs (mtDNAs) are typically single circular chromosomes, with the exception of those from medusozoan cnidarians (jellyfish and hydroids), which are linear and sometimes fragmented. Most medusozoans have linear monomeric or linear bipartite mitochondrial genomes, but preliminary data have suggested that box jellyfish (cubozoans) have mtDNAs that consist of many linear chromosomes. Here, we present the complete mtDNA sequence from the winged box jellyfish Alatina moseri (the first from a cubozoan). This genome contains unprecedented levels of fragmentation: 18 unique genes distributed over eight 2.9- to 4.6-kb linear chromosomes. The telomeres are identical within and between chromosomes, and recombination between subtelomeric sequences has led to many genes initiating or terminating with sequences from other genes (the most extreme case being 150 nt of a ribosomal RNA containing the 5′ end of nad2), providing evidence for a gene conversion–based model of telomere evolution. The silent-site nucleotide variation within the A. moseri mtDNA is among the highest observed from a eukaryotic genome and may be associated with elevated rates of recombination

The Telomere Binding Protein TRF2 Induces Chromatin Compaction

Mammalian telomeres are specialized chromatin structures that require the telomere binding protein, TRF2, for maintaining chromosome stability. In addition to its ability to modulate DNA repair activities, TRF2 also has direct effects on DNA structure and topology. Given that mammalian telomeric chromatin includes nucleosomes, we investigated the effect of this protein on chromatin structure. TRF2 bound to reconstituted telomeric nucleosomal fibers through both its basic N-terminus and its C-terminal DNA binding domain. Analytical agarose gel electrophoresis (AAGE) studies showed that TRF2 promoted the folding of nucleosomal arrays into more compact structures by neutralizing negative surface charge. A construct containing the N-terminal and TRFH domains together altered the charge and radius of nucleosomal arrays similarly to full-length TRF2 suggesting that TRF2-driven changes in global chromatin structure were largely due to these regions. However, the most compact chromatin structures were induced by the isolated basic N-terminal region, as judged by both AAGE and atomic force microscopy. Although the N-terminal region condensed nucleosomal array fibers, the TRFH domain, known to alter DNA topology, was required for stimulation of a strand invasion-like reaction with nucleosomal arrays. Optimal strand invasion also required the C-terminal DNA binding domain. Furthermore, the reaction was not stimulated on linear histone-free DNA. Our data suggest that nucleosomal chromatin has the ability to facilitate this activity of TRF2 which is thought to be involved in stabilizing looped telomere structures

Surface acoustic wave excitation on SF6 plasma treated AlGaN/GaN heterostructure

ISBN : 978-1-4244-2325-5International audienceWe present a new modified approach in the forming of interdigital transducers (IDTs) on AlGaN/GaN heterostructure fully compatible with the process technology of HEMTs. The modified approach uses a shallow recess-gate plasma etching of AlGaN barrier layer in combination with "in-situ" SF6 surface plasma treatment under Schottky gate fingers of IDTs. It enables to excite surface acoustic wave (SAW) in the AlGaN/GaN heterostructure without any high external bias voltages needed to apply to the IDTs. The initial results in the process technology and characterization are presented

Surface Acoustic Wave Excitation on SF6 plasma treated AlGaN/GaN heterostructure

Nowadays, there is an increasing interest to generate the Surface Acoustic Wave (SAW) directly on AlGaN/GaN heterostructure material system that defines a two-dimensional electron gas (2DEG) of the high electron mobility transistors (HEMTs). However, the direct interaction of SAW with free carriers of 2DEG prevents the acousto-electric transduction in the interdigital transducers (IDTs) due to the additional insertion losses. Therefore, in this work, we present a new approach in forming of interdigital (SAW) structures on AlGaN/GaN heterostructure to be applied for chemical sensors technology. The new approach uses a selective self-aligned SF6 plasma treatment of the AlGaN/GaN heterostructure to modify 2DEG density thus enabling the redistribution of the E-field and reduction of the peak field in the range of SAW structures. It enables to control these losses, so SAW propagation in the AlGaN/GaN can be observed

Surface Acoustic Wave Excitation on SF6 plasma treated AlGaN/GaN heterostructure

International audienceIn this work, we introduce a new modified approach to the formation of interdigital transducer (IDT) structures on an AlGaN/GaN heterostructure. The approach is based on a shallow recess-gate plasma etching of the AlGaN barrier layer in combination with “in-situ” SF6 surface plasma treatment applied selectively under the Schottky gate fingers of IDTs. It enables one to modify the two-dimensional electron gas (2DEG) density and the surface field distribution in the region of the IDTs, as is needed for the excitation of a surface acoustic wave (SAW). The measured transfer characteristics of the plasma treated SAW structures revealed the excitation of SAW at zero bias voltage due to fully depleted 2DEG in the region of the IDTs. High external bias voltages are not necessary for SAW excitation. SIMS depth distribution profiles of F atoms were measured to discuss the impact of SF6 plasma treatment on the performance of the AlGaN/GaN-based IDTs

Liquid-liquid extraction of biomolecules in downstream processing - A review paper

Economic analysis shows that protein separation and purification are a very important aspect of biomolecules production and processing. This is particularly true for protein processing which, because of the complexity of the starting material, often requires many steps to reach the levels of purity required for medical and food applications. The separation specialists' task is to develop safe and simple processes to achieve products with a high level of purity. On a large scale, chromatography of proteins is not an easily applied method, although on a laboratory scale it is very effective and relatively simple. When it is scaled up, shortcomings such as discontinuity in the process, slow protein diffusion and large pressure drops in the system are seen. For these reasons a substantial research effort has been directed toward the use of aqueous two-phase systems (ATPSs) to replace the initial steps in protein purification and chromatography. This article reviews the chronology and main ATPS fundamentals and discuss the broader applications of this type of system in the extraction and separation of biomolecules