Post-translational regulation of lipophorin receptors

Resumen del póster presentado al IX Meeting of the Spanish Society for Developmental Biology celebrado en Granada del 12 al 14 de noviembre de 2012.Peer Reviewe

Managing the business operations of a travel agency

U ovome radu obavljeno je istraživanje tržišta za online korištenje usluga putničkih agencija. Navedene su neke od stranica za mrežnu kupovinu kao primjeri, alati u kojima su izrađene te je opisana njihova funkcionalnosti i nedostaci. Pojašnjeni su alati korišteni tijekom izrade rada: SQL Workbench i SQL Server u svrhu izrade i povezivanja s bazom i Visual Studio za izradu mrežne aplikacije. Temeljito je obrađena baza podataka s osvrtom na ER dijagram koja je implementirana u navedenu aplikaciju. Pojašnjene su namjene kreiranih tablica i njihovih atributa kao i vrste relacijskih veza među njima. Navedene su sve mogućnosti i ograničenja korištenja ovog relacijskog modela. Posebna se pažnja posvetila sigurnosti baze od namjernih ili slučajnih pristupa podacima. U teorijskome se dijelu opisuje najčešće korištena metoda za neovlašten pristup te su spomenuti načini na koje se to može spriječiti. Opisan je i postupak za provjeru sigurnosti vlastite baze. U praktičnom djelu rada pomoću ASP.NET MVC tehnologije je izrađena aplikacija pisana u programskom jeziku C#. Ona se sastoji od odvojenih sučelja za administratora i klijente. Pošto svaka agencija upravlja povjerljivim informacijama, navedena podjela ima ograničeni pristup podacima osobama koje za to nisu ovlaštene. Administrator ima dozvolu za potpuno upravljanje radom agencije: pregled ponude, klijentele, ostvarenih rezervacija i računa te dodavanje, promjenu ili brisanje navedenih stavki. Klijentima je dopušten pregled ponude i ostvarivanje rezervacija ili uplata za željen turistički aranžman. U radu su opisane korištene metode, postupci i načini funkcioniranja. Dati su osnovni elementi dizajna i funkcionalnosti svakog pripadajućeg segmenta za potrebe kvalitetnog vođenja putničke agencije. Grafičko sučelje je intuitivno i prilagodljivo svakom korisniku te bez gubitka funkcionalnosti što je prikazano na priloženim slikama u ovome radu

Hydrogeological impact assessment by tunnelling at sites of high sensitivity

A tunnel for the High Speed Train (HST) was constructed in Barcelona with an Earth Pressure Balance (EPB) Tunnel Boring Machine (TBM). The tunnel crosses Barcelona and passes under some famous landmarks such as the Sagrada Familia and the Casa Mill Both monuments are UNESCO world heritage sites and a committee appointed by the UNESCO acted as external observers during the construction. Concerns about soil settlements and the hydrogeological impacts of the construction were raised. These concerns were addressed during the design stage to forestall any unexpected events. The methodology consisted of 1) characterising the geology in detail, 2) predicting the impacts caused in the aquifer, 3) predicting the soil displacements due to water table oscillations produced by the construction, and 4) monitoring the evolution of groundwater and soil settlements. The main estimated impact on groundwater was a moderate barrier effect. The barrier effect, the magnitude of which matched the predictions, was detected during construction. The monitoring of soil settlements revealed short and long term movements. The latter movements matched the analytical predictions of soil displacements caused by the groundwater oscillations. This paper proposes a realistic procedure to estimate impacts on groundwater during tunnel construction with an EPB. Our methodology will considerably improve the construction of tunnels in urban areas. (C) 2015 Elsevier B.V. All rights reserved.Peer ReviewedPostprint (author's final draft

AEBP2 as a potential targeting protein for Polycomb Repression Complex PRC2

AEBP2 is a zinc finger protein that has been shown to interact with the mammalian Polycomb Repression Complex 2 (PRC2). In the current study, we characterized this unknown protein and tested its potential targeting roles for the PRC2. AEBP2 is an evolutionarily well-conserved gene that is found in the animals ranging from flying insects to mammals. The transcription of mammalian AEBP2 is driven by two alternative promoters and produces at least two isoforms of the protein. These isoforms show developmental stage-specific expression patterns: the adult-specific larger form (51 kDa) and the embryo-specific smaller form (32 kDa). The AEBP2 protein binds to a DNA-binding motif with an unusual bipartite structure, CTT(N)15-23cagGCC with lower-case being less critical. A large fraction of AEBP2's target loci also map closely to the known target loci of the PRC2. In fact, many of these loci are co-occupied by the two proteins, AEBP2 and SUZ12. This suggests that AEBP2 is most likely a targeting protein for the mammalian PRC2 complex

Global membrane protein interactome analysis using <i>in vivo</i> crosslinking and MS-based protein correlation profiling

We present a methodology using in vivo crosslinking combined with HPLC-MS for the global analysis of endogenous protein complexes by protein correlation profiling. Formaldehyde crosslinked protein complexes were extracted with high yield using denaturing buffers that maintained complex solubility during chromatographic separation. We show this efficiently detects both integral membrane and membrane-associated protein complexes,in addition to soluble complexes, allowing identification and analysis of complexes not accessible in native extracts. We compare the protein complexes detected by HPLC-MS protein correlation profiling in both native and formaldehyde crosslinked U2OS cell extracts. These proteome-wide data sets of both in vivo crosslinked and native protein complexes from U2OS cells are freely available via a searchable online database (www.peptracker.com/epd). Raw data are also available via ProteomeXchange (identifier PXD003754)

Negative Regulation of EGFR/MAPK Pathway by Pumilio in Drosophila melanogaster

In Drosophila melanogaster, specification of wing vein cells and sensory organ precursor (SOP) cells, which later give rise to a bristle, requires EGFR signaling. Here, we show that Pumilio (Pum), an RNA-binding translational repressor, negatively regulates EGFR signaling in wing vein and bristle development. We observed that loss of Pum function yielded extra wing veins and additional bristles. Conversely, overexpression of Pum eliminated wing veins and bristles. Heterozygotes for Pum produced no phenotype on their own, but greatly enhanced phenotypes caused by the enhancement of EGFR signaling. Conversely, over-expression of Pum suppressed the effects of ectopic EGFR signaling. Components of the EGFR signaling pathway are encoded by mRNAs that have Nanos Response Element (NRE)–like sequences in their 3’UTRs; NREs are known to bind Pum to confer regulation in other mRNAs. We show that these NRE-like sequences bind Pum and confer repression on a luciferase reporter in heterologous cells. Taken together, our evidence suggests that Pum functions as a negative regulator of EGFR signaling by directly targeting components of the pathway in Drosophila

Two or Four Bristles: Functional Evolution of an Enhancer of scute in Drosophilidae

Changes in cis-regulatory sequences are proposed to underlie much of morphological evolution. Yet, little is known about how such modifications translate into phenotypic differences. To address this problem, we focus on the dorsocentral bristles of Drosophilidae. In Drosophila melanogaster, development of these bristles depends on a cis-regulatory element, the dorsocentral enhancer, to activate scute in a cluster of cells from which two bristles on the posterior scutum arise. A few species however, such as D. quadrilineata, bear anterior dorsocentral bristles as well as posterior ones, a derived feature. This correlates with an anterior expansion of the scute expression domain. Here, we show that the D. quadrilineata enhancer has evolved, and is now active in more anterior regions. When used to rescue scute expression in transgenic D. melanogaster, the D. quadrilineata enhancer is able to induce anterior bristles. Importantly, these properties are not displayed by homologous enhancers from control species bearing only two posterior bristles. We also provide evidence that upstream regulation of the enhancer, by the GATA transcription factor Pannier, has been evolutionarily conserved. This work illustrates how, in the context of a conserved trans-regulatory landscape, evolutionary tinkering of pre-existing enhancers can modify gene expression patterns and contribute to morphological diversification

Crosstalk between the EGFR and other signalling pathways at the level of the global transcriptional corepressor Groucho/TLE

In this minireview, we briefly revisit the Drosophila Notch and epidermal growth factor receptor pathways, and relate to the relationship between them. We then mainly focus on the involvement of Groucho (Gro)/TLE, a global developmental corepressor, in these pathways. In particular, we discuss Gro/TLE's role at the junction between these two signal transduction cascades

NF-κB/Rel-Mediated Regulation of the Neural Fate in Drosophila

Two distinct roles are described for Dorsal, Dif and Relish, the three NF-κB/Rel proteins of Drosophila, in the development of the peripheral nervous system. First, these factors regulate transcription of scute during the singling out of sensory organ precursors from clusters of cells expressing the proneural genes achaete and scute. This effect is possibly mediated through binding sites for NF-κB/Rel proteins in a regulatory module of the scute gene required for maintenance of scute expression in precursors as well as repression in cells surrounding precursors. Second, genetic evidence suggests that the receptor Toll-8, Relish, Dif and Dorsal, and the caspase Dredd pathway are active over the entire imaginal disc epithelium, but Toll-8 expression is excluded from sensory organ precursors. Relish promotes rapid turnover of transcripts of the target genes scute and asense through an indirect, post-transcriptional mechanism. We propose that this buffering of gene expression levels serves to keep the neuro-epithelium constantly poised for neurogenesis

Control of Neural Daughter Cell Proliferation by Multi-level Notch/Su(H)/E(spl)-HLH Signaling

The Notch pathway controls proliferation during development and in adulthood, and is frequently affected in many disorders. However, the genetic sensitivity and multi-layered transcriptional properties of the Notch pathway has made its molecular decoding challenging. Here, we address the complexity of Notch signaling with respect to proliferation, using the developing Drosophila CNS as model. We find that a Notch/Su(H)/E(spl)-HLH cascade specifically controls daughter, but not progenitor proliferation. Additionally, we find that different E(spl)-HLH genes are required in different neuroblast lineages. The Notch/Su(H)/E(spl)-HLH cascade alters daughter proliferation by regulating four key cell cycle factors: Cyclin E, String/Cdc25, E2f and Dacapo (mammalian p21CIP1/p27KIP1/p57Kip2). ChIP and DamID analysis of Su(H) and E(spl)-HLH indicates direct transcriptional regulation of the cell cycle genes, and of the Notch pathway itself. These results point to a multi-level signaling model and may help shed light on the dichotomous proliferative role of Notch signaling in many other systems