ELKS/Voltage-Dependent Ca2+ Channel-β Subunit Module Regulates Polarized Ca2+ Influx in Pancreatic β Cells

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Genome-wide localization of pre-RC sites and identification of replication origins in fission yeast

DNA replication of eukaryotic chromosomes initiates at a number of discrete loci, called replication origins. Distribution and regulation of origins are important for complete duplication of the genome. Here, we determined locations of Orc1 and Mcm6, components of pre-replicative complex (pre-RC), on the whole genome of Schizosaccharomyces pombe using a high-resolution tiling array. Pre-RC sites were identified in 460 intergenic regions, where Orc1 and Mcm6 colocalized. By mapping of 5-bromo-2'-deoxyuridine (BrdU)-incorporated DNA in the presence of hydroxyurea (HU), 307 pre-RC sites were identified as early-firing origins. In contrast, 153 pre-RC sites without BrdU incorporation were considered to be late and/or inefficient origins. Inactivation of replication checkpoint by Cds1 deletion resulted in BrdU incorporation with HU specifically at the late origins. Early and late origins tend to distribute separately in large chromosome regions. Interestingly, pericentromeric heterochromatin and the silent mating-type locus replicated in the presence of HU, whereas the inner centromere or subtelomeric heterochromatin did not. Notably, MCM did not bind to inner centromeres where origin recognition complex was located. Thus, replication is differentially regulated in chromosome domains.     

Long-lasting effects of sedentarization-induced increase of fertility on labor force proportion in an Arab socieity: a case study in south Jordan

Based on the authors' genealogical-demographic data of a Bedouin-origin clan in south Jordan which settled down in 1948, this paper aimed to report the long-lasting effects of increase in fertility induced by sedentarization on the labor force proportion, defined as percent of persons aged 15-64 years. The results highlighted that this proportion had an inverse relation with the total fertility rate and tended to fluctuate up to the present time with an interval of several decades, suggesting that a Bedouin-origin sedentarized group met alternately with the phase of high fertility and low proportion of labor force and the other phase of an opposite pattern. At present, unemployment of males, who have been engaged in subsistence and cash-earning work, was high especially in younger groups basically due to the narrow land of the clan (only 35 hectares for 456 people in 2005) for agriculture and the limited opportunities for cash-earning jobs. Taking into consideration that Bedouin-origin people sedentarized according to the government policy, it is needed for the government to support rural development plans of sedentarized Bedouin peoples.     

Identification of a fatty acyl-CoA synthetase gene, lcf2+, which affects viability after entry into the stationary phase in Schizosaccharomyces pombe

The lcf1(+) gene, which encodes a long chain fatty acyl-CoA synthetase, is necessary for the maintenance of viability after entry into the stationary phase in Schizosaccharomyces pombe. In this study, we analyzed a paralogous gene, SPBP4H10.11c (named lcf2(+)), and we present evidence that the gene encodes a new fatty acyl-CoA synthetase. The enzyme preferentially recognized myristic acid as a substrate. A Deltalcf2 mutant showed increased viability after entry into the stationary phase in SD medium. A Deltalcf1Deltalcf2 double mutant showed a severe decrease in long-chain fatty acyl-CoA synthetase activity and a rapid loss of viability after entry into the stationary phase. These results suggest that fatty acid utilization and/or metabolism is important to determine viability in the stationary phase.     

Inhibition of the gene expression for granule-bound starch synthase I by RNA interference in sweet potato plants

Granule-bound starch synthase I (GBSSI) is one of the key enzymes catalyzing the formation of amylose, a linear α(1,4)D-glucan polymer, from ADP-glucose. Amylose-free transgenic sweet potato plants were produced by inhibiting sweet potato GBSSI gene expression through RNA interference. The gene construct consisting of an inverted repeat of the first exon separated by intron 1 of GBSSI driven by the CaMV 35S promoter was integrated into the sweet potato genome by Agrobacterium tumefaciens-mediated transformation. In over 70% of the regenerated transgenic plants, the expression of GBSSI was inactivated giving rise to storage roots containing amylopectin but not amylose. Electrophoresis analysis failed to detect the GBSSI protein, suggesting that gene silencing of the GBSSI gene had occurred. These results clearly demonstrate that amylose synthesis is completely inhibited in storage roots of sweet potato plants by the constitutive production of the double-stranded RNA of GBSSI fragments. We conclude that RNA interference is an effective method for inhibiting gene expression in the starch metabolic pathway.     

The lectin domains of polypeptide GalNAc-transferases exhibit carbohydrate-binding specificity for GalNAc: lectin binding to GalNAc-glycopeptide substrates is required for high density GalNAc-O-glycosylation

Initiation of mucin-type O-glycosylation is controlled by a large family of UDP GalNAc:polypeptide N-acetylgalactosaminyltransferases (GalNAc-transferases). Most GalNAc-transferases contain a ricin-like lectin domain in the C-terminal end, which may confer GalNAc-glycopeptide substrate specificity to the enzyme. We have previously shown that the lectin domain of GalNAc-T4 modulates its substrate specificity to enable unique GalNAc-glycopeptide specificities and that this effect is selectively inhibitable by GalNAc; however, direct evidence of carbohydrate binding of GalNAc-transferase lectins has not been previously presented. Here, we report the direct carbohydrate binding of two GalNAc-transferase lectin domains, GalNAc-T4 and GalNAc-T2, representing isoforms reported to have distinct glycopeptide activity (GalNAc-T4) and isoforms without apparent distinct GalNAc-glycopeptide specificity (GalNAc-T2). Both lectins exhibited specificity for binding of free GalNAc. Kinetic and time-course analysis of GalNAc-T2 demonstrated that the lectin domain did not affect transfer to initial glycosylation sites, but selectively modulated velocity of transfer to subsequent sites and affected the number of acceptor sites utilized. The results suggest that GalNAc-transferase lectins serve to modulate the kinetic properties of the enzymes in the late stages of the initiation process of O-glycosylation to accomplish dense or complete O-glycan occupancy.     

Effect of waterfalls on fluvial fish distribution and landlocked Rhinogobius brunneus populations on Yakushima Island,Japan

The distribution of fluvial fish was surveyed at 55 sites on Yakushima Island, Japan, which has precipitous mountains and waterfalls (below- or no-waterfall sites: 31; above-waterfall sites: 24). Eleven diadromous and one river resident (introduced Oncorhynchus masou masou) species were found, but absolutely no fish were detected at the 18 above-waterfall sites. Statistical analyses revealed that the presence of waterfalls (> 5 m in height) below the sites had a significant negative effect on fish distribution, suggesting that waterfalls prevent migration of diadromous fishes. We found Rhinogobius brunneus populations above high waterfalls such as Nunobiki Falls (50 m) and Ohko Falls (88 m). Otolith Sr:Ca ratios and mitochondrial DNA (cyt-b region) were examined to determine the migratory history and genetic properties of these populations; the Sr:Ca ratios indicated that the populations had a landlocked life cycle, whereas the genetic endemism/isolation of landlocked populations was unexpectedly absent. There is no clear explanation for this phenomenon, but fairly infrequent individuals of nonlandlocked type might have migrated beyond the waterfalls with their sucker-like organ and mated with landlocked populations, disrupting the genetic isolation of landlocked populations.     

A new species of Gastrosaccus Norman, 1868 (Mysida,Mysidae, Gastrosaccinae) from a sandy shore of Indonesia

A new species of the mysid crustacean genus Gastrosaccus Norman, 1868 (Mysida, Mysidae, Gastrosaccinae) is reported from a sandy shore of Lombok Island, Indonesia. These specimens resemble G. sorrentoensis Wooldridge & McLachlan, 1986 and G. yuyu Bamber and Morton, 2012 by the possession of an articulated process on the fifth abdominal somite together with a fringe of spine-like filaments on the posterodorsal margin of the carapace. The Lombok population differs from the known congeners by having comparatively fewer numbers of carpopropod segments on the endopod of the third to eighth thoracic limbs and the conformation in the telson and in the male third pleopod. Hence, G. lombokiensis sp. n. is proposed herein as a third species of “G. sorrentoensis” species group.     

Mechanism-based inhibition of cancer metastasis with (−)-epigallocatechin gallate

Cell motility and cell stiffness are closely related to metastatic activity of cancer cells. (−)-Epigallocatechin gallate (EGCG) has been shown to inhibit spontaneous metastasis of melanoma cell line into the lungs of mice, so we studied the effects of EGCG on cell motility, cell stiffness, and expression of vimentin and Slug, which are molecular phenotypes of epithelial–mesenchymal transition (EMT). Treatments of human non-small cell lung cancer cell lines H1299 and Lu99 with 50 and 100 μM EGCG reduced cell motility to 67.5% and 43.7% in H1299, and 71.7% and 31.5% in Lu99, respectively in in vitro wound healing assay. Studies on cell stiffness using atomic force microscope (AFM) revealed that treatment with 50 μM EGCG increased Young’s modulus of H1299 from 1.24 to 2.25 kPa and that of Lu99 from 1.29 to 2.28 kPa, showing a 2-fold increase in cell stiffness, i.e. rigid elasticity of cell membrane. Furthermore, treatment with 50 μM EGCG inhibited high expression of vimentin and Slug in the cells at a leading edge of scratch. Methyl-β-cyclodextrin, a reagent to deplete cholesterol in plasma membrane, showed inhibition of EMT phenotypes similar that by EGCG, suggesting that EGCG induces inhibition of EMT phenotypes by alteration of membrane organization.     

On the foraging and feeding ability of Oithona davisae (Crustacea,Copepoda)

Growth experiments with macrophytes revealed anomalies in the growth and development of Najas intermedia. Further analyses showed that these anomalies were caused by differences between the male and female plants. To examine the gender-related differences, the results of two field experiments and a phenological monitoring programme were analysed. The field experiments were conducted in four different lakes in Southern Germany. Over the course of 2 years, eight different sediments were used to research the interaction between the gender and the growth of N. intermedia plants. The phenological monitoring programme, on the other hand, was focused on the development of three different populations of N. intermedia in three Southern German lakes. The results of the experiments revealed significant differences in the gender-related development: Depending on the environmental conditions, male plants grew between 20 and 40% faster than female plants. Additionally, the male plants developed flowers earlier in the vegetation period and died before the female plants. As a result, male and female plants coexisted in the same lake only for a short period of time. This might be a strategy to reduce the competitive pressure between the genders and to gain an advantage over rival species.