Role of the ubiquitin-like protein Hub1 in splice-site usage and alternative splicing

Alternative splicing of pre-messenger RNAs diversifies gene products in eukaryotes and is guided by factors that enable spliceosomes to recognize particular splice sites. Here we report that alternative splicing of Saccharomyces cerevisiae SRC1 pre-mRNA is promoted by the conserved ubiquitin-like protein Hub1. Structural and biochemical data show that Hub1 binds non-covalently to a conserved element termed HIND, which is present in the spliceosomal protein Snu66 in yeast and mammals, and Prp38 in plants. Hub1 binding mildly alters spliceosomal protein interactions and barely affects general splicing in S. cerevisiae. However, spliceosomes that lack Hub1, or are defective in Hub1-HIND interaction, cannot use certain non-canonical 5' splice sites and are defective in alternative SRC1 splicing. Hub1 confers alternative splicing not only when bound to HIND, but also when experimentally fused to Snu66, Prp38, or even the core splicing factor Prp8. Our study indicates a novel mechanism for splice site utilization that is guided by non-covalent modification of the spliceosome by an unconventional ubiquitin-like modifier.     

Linear and nonlinear optical changes in amorphous As2Se3 thin film upon UV exposure

Amorphous As_2Se_3 chalcogenide thin film was exposed to UV light(i line and g line) using mercury lamp for 30 min. XRD,UV/VIS spectroscopy and thickness measurements were taken for unexposed and exposed thin film for structural and optical characterizations.Linear optical constants(linear refractive index,extinction coefficient and linear optical absorption coefficient) of the film were calculated from transmission spectra using Swanepoel method.Optical bandgap was determined using Tauc's relation of in...     

The rapid drift of the Indian tectonic plate

The breakup of the supercontinent Gondwanaland into Africa, Antarctica, Australia and India about 140 million years ago, and consequently the opening of the Indian Ocean, is thought to have been caused by heating of the lithosphere from below by a large plume whose relicts are now the Marion, Kerguelen and Réunion plumes. Plate reconstructions based on palaeomagnetic data suggest that the Indian plate attained a very high speed (18-20 cm yr(-1) during the late Cretaceous period) subsequent to its breakup from Gondwanaland, and then slowed to approximately 5 cm yr(-1) after the continental collision with Asia approximately 50 Myr ago. The Australian and African plates moved comparatively less distance and at much lower speeds of 2-4 cm yr(-1) (refs 3-5). Antarctica remained almost stationary. This mobility makes India unique among the fragments of Gondwanaland. Here we propose that when the fragments of Gondwanaland were separated by the plume, the penetration of their lithospheric roots into the asthenosphere were important in determining their speed. We estimated the thickness of the lithospheric plates of the different fragments of Gondwanaland around the Indian Ocean by using the shear-wave receiver function technique. We found that the fragment of Gondwanaland with clearly the thinnest lithosphere is India. The lithospheric roots in South Africa, Australia and Antarctica are between 180 and 300 km deep, whereas the Indian lithosphere extends only about 100 km deep. We infer that the plume that partitioned Gondwanaland may have also melted the lower half of the Indian lithosphere, thus permitting faster motion due to ridge push or slab pull.     

Transvascular delivery of small interfering RNA to the central nervous system

A major impediment in the treatment of neurological diseases is the presence of the blood-brain barrier, which precludes the entry of therapeutic molecules from blood to brain. Here we show that a short peptide derived from rabies virus glycoprotein (RVG) enables the transvascular delivery of small interfering RNA (siRNA) to the brain. This 29-amino-acid peptide specifically binds to the acetylcholine receptor expressed by neuronal cells. To enable siRNA binding, a chimaeric peptide was synthesized by adding nonamer arginine residues at the carboxy terminus of RVG. This RVG-9R peptide was able to bind and transduce siRNA to neuronal cells in vitro, resulting in efficient gene silencing. After intravenous injection into mice, RVG-9R delivered siRNA to the neuronal cells, resulting in specific gene silencing within the brain. Furthermore, intravenous treatment with RVG-9R-bound antiviral siRNA afforded robust protection against fatal viral encephalitis in mice. Repeated administration of RVG-9R-bound siRNA did not induce inflammatory cytokines or anti-peptide antibodies. Thus, RVG-9R provides a safe and noninvasive approach for the delivery of siRNA and potentially other therapeutic molecules across the blood-brain barrier.     

A preliminary report on gamma-irradiated protein-dye complexes

In spectral studies of gamma-irradiated protein-dye complexes, influences of concentrations of the components and changes in dye character are mainly noted.     

Cytoprotective effects of acidosis via heat shock protein HSP27 against the anticancer drug doxorubicin

Drug resistance continues to be a stumbling block in achieving better cure rates in several cancers. Doxorubicin is commonly used in treatment of a wide range of cancers. The aim of this study was to look into the mechanisms of how low ambient pH may contribute to down-regulation of apoptotic pathways in a gastric tumour cell line. Low pH culture conditions were found to dramatically prolong cell survival after doxorubicin treatment, an effect that was in part reversed by co-incubation with the specific p38 mitoge-activated protein kinase (MAP kinase) inhibitor SB203580, only mildly inhibited by blockade of the multi-drug resistance 1 (MDR1) transporter, but completely abolished by siRNA-mediated knockdown of the heat shock protein 27 (HSP27). In conclusion, acidic pH causes less accumulation of cytotoxic drug in the nucleus of adeno gastric carcinoma (AGS) cells and HSP27-dependent decrease in FasR-mediated gastric epithelial tumour cell apoptosis.     

An Architecture of Computer Aided Process Planning System Integrated with Scheduling Using Decision Support System

Process planning and scheduling are two major plann in g and control activities that consume significant part of the lead-time, theref ore all attempts are being made to reduce lead-time by automating them. Compute r Aided Process Planning (CAPP) is a step in this direction. Most of the existin g CAPP systems do not consider scheduling while generating a process plan. Sched uling is done separately after the process plan has been generated and therefore , it is possible that a process plan so generated is e...     

In vitro demonstration of peroxidase activity in the fish kidney soluble supernatant and its physiological importance

Zusammenfassung In der Kopfniere des TeleostiersAnabes testudineus wurde eine besonders hohe Peroxidase-Aktivität nachgewiesen, insbesondere deren Iodid-Oxydation zu Tri-Iodid.     

Cytotoxic T cells specific for the circumsporozoite protein of Plasmodium falciparum

Malaria is initiated by the inoculation of a susceptible host with sporozoites from an infected mosquito. The sporozoites enter hepatocytes and develop for a period as exoerythrocyte or hepatic stage parasites. Vaccination with irradiated sporozoites can provide protective immunity and a recent study shows that this can also be conferred by immunization with a recombinant salmonella expressing only the circumsporozoite protein that normally covers the sporozoites. Protection against infection is likely to be mediated by cytotoxic CD8+ cells, as depletion of CD8+ T cells in a sporozoite-immunized animal can completely abrogate immunity. Here we demonstrate directly the existence of CD8+ cytotoxic T lymphocytes (CTL) that recognize the circumsporozoite protein. B10.BR mice immunized with sporozoites or with recombinant vaccinia virus expressing the CS protein of Plasmodium falciparum contain CTL that specifically kill L cell fibroblasts transfected with the gene encoding the same CS protein. The peptide epitope from the CS protein that is recognized by CTL from this strain of mice is from a variant region of the protein.