Expression and Visualization of Red Fluorescent Protein (RFP) in Neurospora crassa

We report the expression of Discosoma red fluorescent protein (RFP) and RFP fusion proteins in Neurospora crassa. RFP was expressed under the control of the Neurospora ccg-1 promoter in transformants with single copies integrated at the his-3 locus by gene targeting. Because this RFP gene, tdimer2(12), contains a 677 bp direct tandem repeat of dsRed, RFP constructs underwent RIP at high frequency in rid+ strains. Fusion proteins of RFP to the amino terminus of Neurospora heterochromatin protein 1 (HP1) were localized to heterochromatic foci in Neurospora nuclei, consistent with prior findings with carboxy-terminal HP1-GFP fusion proteins

The Significance of Studying a Trade Diaspora

Situated halfway between Aden and Oman, the Yemeni province of Hadhramaut is considered by most fleeting visitors a backwater, notable only for the highrise mudbrick houses of the former trading centre of Shibam and the extravagant but decaying palaces of neighbouring Say'un and Tarim. Little is known, however, about the people who built these remarkable constructions, and about their far-reaching connections in the areas bordering on the Indian Ocean. However, their story, if recovered, sheds light on a number of questions pertinent to current interests in Middle Eastern and Islamic history. Let us consider the biography of one such trader, whose cosmopolitanism in entrepreneurial, political, and intellectual terms is quite typical for a wider group of Hadhramis, as well as probably for members of other such groups in the Indian Ocean and beyond

Improved plasmids for gene targeting at the his-3 locus of Neurospora crassa by electroporation: Correction

Two mistakes in our article on gene replacement by gene targeting at the his-3 locus (Margolin, B.S. et al., 1997, FGN 44:34-36) have come to our attention

Expression and visualization of Green Fluorescent Protein (GFP) in Neurospora crassa

We report the first successful imaging of GFP expression in Neurospora crassa. GFP was expressed under the control of the heterologous ToxA promoter from Pyrenophora tritici-repentis in transformants carrying multiple or single copies of the GFP construct. GFP was also detected in ascospores but not during earlier stages of the sexual cycle

Improved plasmids for gene targeting at the his-3 locus of Neurospora crassa by electroporation

We report two new plasmids, pBM60 and pBM61, and procedures to efficiently generate single- copy transformants targeted to the his-3 locus in Neurospora crassa

Nanolithography and manipulation of graphene using an atomic force microscope

We use an atomic force microscope (AFM) to manipulate graphene films on a nanoscopic length scale. By means of local anodic oxidation with an AFM we are able to structure isolating trenches into single-layer and few-layer graphene flakes, opening the possibility of tabletop graphene based device fabrication. Trench sizes of less than 30 nm in width are attainable with this technique. Besides oxidation we also show the influence of mechanical peeling and scratching with an AFM of few layer graphene sheets placed on different substrates.Comment: 11 pages text, 5 figure

Influence of a stellar cusp on the dynamics of young stellar discs and the origin of the S-stars in the Galactic Centre

Observations of the Galactic Centre show evidence of one or two disc-like structures of very young stars orbiting the central super-massive black hole within a distance of a few 0.1 pc. A number of analyses have been carried out to investigate the dynamical behaviour and consequences of these discs, including disc thickness and eccentricity growth as well as mutual interaction and warping. However, most of these studies have neglected the influence of the stellar cusp surrounding the black hole, which is believed to be 1-2 orders of magnitude more massive than the disc(s). By means of N-body integrations using our bhint code, we study the impact of stellar cusps of different compositions. We find that although the presence of a cusp does have an important effect on the evolution of an otherwise isolated flat disc, its influence on the evolution of disc thickness and warping is rather mild in a two-disc configuration. However, we show that the creation of highly eccentric orbits strongly depends on the graininess of the cusp (i.e. the mean and maximum stellar masses): While Chang (2009) recently found that full cycles of Kozai resonance are prevented by the presence of an analytic cusp, we show that relaxation processes play an important role in such highly dense regions and support short-term resonances. We thus find that young disc stars on initially circular orbits can achieve high eccentricities by resonant effects also in the presence of a cusp of stellar remnants, yielding a mechanism to create S-stars and hyper-velocity stars. Furthermore, we discuss the underlying initial mass function (IMF) of the young stellar discs and find no definite evidence for a non-canonical IMF.Comment: 10 pages, 7 figures, 1 table, accepted for publication in MNRA

The fungus Neurospora crassa displays telomeric silencing mediated by multiple sirtuins and by methylation of histone H3 lysine 9

<p>Abstract</p> <p>Background</p> <p>Silencing of genes inserted near telomeres provides a model to investigate the function of heterochromatin. We initiated a study of telomeric silencing in <it>Neurospora crassa</it>, a fungus that sports DNA methylation, unlike most other organisms in which telomeric silencing has been characterized.</p> <p>Results</p> <p>The selectable marker, <it>hph</it>, was inserted at the subtelomere of Linkage Group VR in an <it>nst-1 </it>(n<it>eurospora </it>s<it>ir </it>t<it>wo</it>-1) mutant and was silenced when <it>nst-1 </it>function was restored. We show that NST-1 is an H4-specific histone deacetylase. A second marker, <it>bar</it>, tested at two other subtelomeres, was similarly sensitive to <it>nst-1 </it>function. Mutation of three additional SIR2 homologues, <it>nst-2</it>, <it>nst-3 </it>and <it>nst-5</it>, partially relieved silencing. Two genes showed stronger effects: <it>dim-5</it>, which encodes a histone H3 K9 methyltransferase and <it>hpo</it>, which encodes heterochromatin protein-1. Subtelomeres showed variable, but generally low, levels of DNA methylation. Elimination of DNA methylation caused partial derepression of one telomeric marker. Characterization of histone modifications at subtelomeric regions revealed H3 trimethyl-K9, H3 trimethyl-K27, and H4 trimethyl-K20 enrichment. These modifications were slightly reduced when telomeric silencing was compromised. In contrast, acetylation of histones H3 and H4 increased.</p> <p>Conclusion</p> <p>We demonstrate the presence of telomeric silencing in Neurospora and show a dependence on histone deacetylases and methylation of histone H3 lysine 9. Our studies also reveal silencing functions for DIM-5 and HP1 that appear independent of their role in <it>de novo </it>DNA methylation.</p

Biogenesis of mitochondrial porin

We review here the present knowledge about the pathway of import and assembly of porin into mitochondria and compare it to those of other mitochondrial proteins. Porin, like all outer mitochondrial membrane proteins studied so far is made as a precursor without a cleavble lsquosignalrsquo sequence; thus targeting information must reside in the mature sequence. At least part of this information appears to be located at the amino-terminal end of the molecule. Transport into mitochondria can occur post-translationally. In a first step, the porin precursor is specifically recognized on the mitochondrial surface by a protease sensitive receptor. In a second step, porin precursor inserts partially into the outer membrane. This step is mediated by a component of the import machinery common to the import pathways of precursor proteins destined for other mitochondrial subcompartments. Finally, porin is assembled to produce the functional oligomeric form of an integral membrane protein wich is characterized by its extreme protease resistance