Hazard index maps for woody material recruitment and transport in alpine catchments

A robust and reliable risk assessment procedure for hydrologic hazards deserves particular attention to the role of transported woody material during flash floods or debris flows. At present, woody material transport phenomena are not systematically considered within the procedures for the elaboration of hazard maps. The consequence is a risk of losing prediction accuracy and of underestimating hazard impacts. Transported woody material frequently interferes with the sediment regulation capacity of open check dams and moreover, when obstruction phenomena at critical crosssections of the stream occur, inundations can be triggered. The paper presents a procedure for the determination of the relative propensity of mountain streams to the entrainment and delivery of recruited woody material on the basis of empirical indicators. The procedure provided the basis for the elaboration of a hazard index map for all torrent catchments of the Autonomous Province of Bolzano/Bozen. The plausibility of the results has been thoroughly checked by a backward oriented analysis on natural hazard events, documented since 1998 at the Department of Hydraulic Engineering of the aforementioned Alpine Province. The procedure provides hints for the consideration of the effects, induced by woody material transport, during the elaboration of hazard zone maps.

The Hydroxyl Side Chain of a Highly Conserved Serine Residue Is Required for Cation Selectivity and Substrate Transport in the Glial Glutamate Transporter GLT-1/SLC1A2

Glutamate transporters maintain synaptic concentration of the excitatory neurotransmitter below neurotoxic levels. Their transport cycle consists of cotransport of glutamate with three sodium ions and one proton, followed by countertransport of potassium. Structural studies proposed that a highly conserved serine located in the binding pocket of the homologous GltPh coordinates l-aspartate as well as the sodium ion Na1. To experimentally validate these findings, we generated and characterized several mutants of the corresponding serine residue, Ser-364, of human glutamate transporter SLC1A2 (solute carrier family 1 member 2), also known as glutamate transporter GLT-1 and excitatory amino acid transporter EAAT2. S364T, S364A, S364C, S364N, and S364D were expressed in HEK cells and Xenopus laevis oocytes to measure radioactive substrate transport and transport currents, respectively. All mutants exhibited similar plasma membrane expression when compared with WT SLC1A2, but substitutions of serine by aspartate or asparagine completely abolished substrate transport. On the other hand, the threonine mutant, which is a more conservative mutation, exhibited similar substrate selectivity, substrate and sodium affinities as WT but a lower selectivity for Na(+) over Li(+). S364A and S364C exhibited drastically reduced affinities for each substrate and enhanced selectivity for l-aspartate over d-aspartate and l-glutamate, and lost their selectivity for Na(+) over Li(+). Furthermore, we extended the analysis of our experimental observations using molecular dynamics simulations. Altogether, our findings confirm a pivotal role of the serine 364, and more precisely its hydroxyl group, in coupling sodium and substrate fluxes.

An environmental magnetic study, sedimentological and numerical methods around Tauranga Harbour, New Zealand

Magnetic iron minerals are widespread and indicative sediment constituents in estuarine, coastal and shelf systems. We combine environmental magnetic, sedimentological and numerical methods to identify magnetite-enriched placer-like zones in a complex coastal system and delineate their formation mechanisms. Magnetic susceptibility and remanence measurements on 245 surficial sediment samples collected in and around Tauranga Harbour, the largest barrier-enclosed tidal estuary of New Zealand, reveal several discrete enrichment zones controlled by local hydrodynamic conditions. Active magnetite enrichment takes place in tidal channels, which feed into two coast-parallel nearshore magnetite-enriched belts centered at water depths of 6-10 m and 10-20 m. A close correlation between magnetite content and magnetic grain size was found, where higher susceptibility values are associated within coarser magnetic crystal sizes. Two key mechanisms for magnetite enrichment are identified. First, tide-induced residual currents primarily enable magnetite enrichment within the estuarine channel network. A coast-parallel, fine sand magnetite enrichment belt in water depths of less than 10 m along the barrier island has a strong decrease in magnetite content away from the southern tidal inlet and is apparently related to active coast-parallel transport combined with mobilizing surf zone processes. A second, less pronounced, but more uniform magnetite enrichment belt at 10-20 m water depth is composed of non-mobile, medium-coarse-grained relict sands, which have been reworked during post-glacial sea level transgression. We demonstrate the potential of magnetic methods to reveal and differentiate coastal magnetite enrichment patterns and investigate their formative mechanisms.

Nonequlibrium particle and energy currents in quantum chains connected to mesoscopic Fermi reservoirs

We propose a model of nonequilibrium quantum transport of particles and energy in a system connected to mesoscopic Fermi reservoirs (mesoreservoir). The mesoreservoirs are in turn thermalized to prescribed temperatures and chemical potentials by a simple dissipative mechanism described by the Lindblad equation. As an example, we study transport in monoatomic and diatomic chains of noninteracting spinless fermions. We show numerically the breakdown of the Onsager reciprocity relation due to the dissipative terms of the model.

Transport and mixing enhancement in fluid-thermal microsystems

En esta tesis se investiga de forma experimental el transporte pasivo de magnitudes físicas en micro-sistemas con carácter de inmediata aplicación industrial, usando métodos innovadores para mejorar la eficiencia de los mismos optimizando parámetros críticos del diseño o encontrar nuevos destinos de posible aplicación. Parte de los resultados obtenidos en estos experimentos han sido publicados en revistas con un índice de impacto tal que pertenecen al primer cuarto del JCR. Primero de todo se ha analizado el efecto que produce en un intercambiador de calor basado en micro-canales el hecho de dejar un espacio entre canales y tapa superior para la interconexión de los mismos. Esto genera efectos tridimensionales que mejoran la exracción de calor del intercambiador y reducen la caída de presión que aparece por el transcurso del fluido a través de los micro-canales, lo que tiene un gran impacto en la potencia que ha de suministrar la bomba de refrigerante. Se ha analizado también la mejora producida en términos de calor disipado de un micro-procesador refrigerado con un ampliamente usado plato de aletas al implementar en éste una cámara de vapor que almacena un fluido bifásico. Se ha desarrollado de forma paralela un modelo numérico para optimizar las nuevas dimensiones del plato de aletas modificado compatibles con una serie de requerimientos de diseño en el que tanto las dimensiones como el peso juegan un papel esencial. Por otro lado, se han estudiado los fenomenos fluido-dinámicos que aparecen aguas abajo de un cuerpo romo en el seno de un fluido fluyendo por un canal con una alta relación de bloqueo. Los resultados de este estudio confirman, de forma experimental, la existencia de un régimen intermedio, caracterizado por el desarrollo de una burbuja de recirculación oscilante entre los regímenes, bien diferenciados, de burbuja de recirculación estacionaria y calle de torbellinos de Karman, como función del número de Reynolds del flujo incidente. Para la obtención, análisis y post-proceso de los datos, se ha contado con la ayuda de un sistema de Velocimetría por Imágenes de Partículas (PIV). Finalmente y como adición a este último punto, se ha estudiado las vibraciones de un cuerpo romo producidas por el desprendimiento de torbellinos en un canal de alta relación de bloqueo con la base obtenida del estudio anterior. El prisma se mueve con un movimiento armónico simple para un intervalo de números de Reynolds y este movimiento se transforma en vibración alrededor de su eje a partir de un ciero número de Reynolds. En relación al fluido, el régimen de desprendimiento de torbellinos se alcanza a menores números de Reynolds que en el caso de tener el cuerpo romo fijo. Uniendo estos dos registros de movimientos y variando la relación de masas entre prisma y fluido se obtiene un mapa con diferentes estados globales del sistema. Esto no solo tiene aplicación como método para promover el mezclado sino también como método para obtener energía a partir del movimiento del cuerpo en el seno del fluido. Abstract In this thesis, experimental research focused on passive scalar transport is performed in micro-systems with marked sense of industrial application, using innovative methods in order to obtain better performances optimizing critical design parameters or finding new utilities. Part of the results obtained in these experiments have been published into high impact factor journals belonged to the first quarter of the Journal Citation Reports (JCR). First of all the effect of tip clearance in a micro-channel based heat sink is analyzed. Leaving a gap between channels and top cover, letting the channels communicate each other causes three-dimensional effects which improve the heat transfer between fluid and heat sink and also reducing the pressure drop caused by the fluid passing through the micro-channels which has a great impact on the total cooling pumping power needed. It is also analyzed the enhancement produced in terms of dissipated heat in a micro-processor cooling system by improving the predominantly used fin plate with a vapour chamber based heat spreader which contains a two-phase fluid inside. It has also been developed at the same time a numerical model to optimize the new fin plate dimensions compatible with a series of design requirements in which both size and wight plays a very restrictive role. On the other hand, fluid-dynamics phenomena that appears downstream of a bluff body in the bosom of a fluid flow with high blockage ratio has been studied. This research experimentally confirms the existence of an intermediate regime characterized by an oscillating closed recirculation bubble intermediate regime between the steady closed recirculation bubble regime and the vortex shedding regime (Karman street like regime) as a function of the incoming flow Reynolds number. A particle image velocimetry technique (PIV) has been used in order to obtain, analyze and post-process the fluid-dynamic data. Finally and as an addition to the last point, a study on the vortexinduced vibrations (VIV) of a bluff body inside a high blockage ratio channel has been carried out taking advantage of the results obtained with the fixed square prism. The prism moves with simple harmonic motion for a Reynolds number interval and this movement becomes vibrational around its axial axis after overcoming at definite Reynolds number. Regarding the fluid, vortex shedding regime is reached at Reynolds numbers lower than the previous critical ones. Merging both movement spectra and varying the square prism to fluid mass ratio, a map with different global states is reached. This is not only applicable as a mixing enhancement technique but as an energy harvesting method.

Beyond the Regional Development Paradigm: The Limits of Transport-Based Policies in Spain

This paper reviews the transport and economic development trends for the last 20 years in Spain at a detailed (province or NUTS3) level. As Spain has sustained a significant transport investment effort in this period, with the support of EU funding, this review offers an excellent perspective to put some further light on how the transport-and-regional-development paradigm has shaped decision-making in the transport sector. The paper reviews changes in gross domestic product (GDP), population and motorway endowment for the 47 provinces in mainland Spain. Regional development trends seem to be closely associated to particular local conditions, not clearly associated to transport (motorway) infrastructure endowment. This is consistent with the fact that transport infrastructure has not generally been a critical bottleneck for trade and economic activity during this period. The paper concludes that, in general terms, transport infrastructure investment does not seem to be clearly associated to the otherwise substantial differences in regional development among Spanish mainland provinces during this period.

Critical radius for hot-jet ignition of hydrogen-air mixtures

This study addresses deflagration initiation of lean and stoichiometric hydrogen–air mixtures by the sudden discharge of a hot jet of their adiabatic combustion products. The objective is to compute the minimum jet radius required for ignition, a relevant quantity of interest for safety and technological applications. For sufficiently small discharge velocities, the numerical solution of the problem requires integration of the axisymmetric Navier–Stokes equations for chemically reacting ideal-gas mixtures, supplemented by standard descriptions of the molecular transport terms and a suitably reduced chemical-kinetic mechanism for the chemistry description. The computations provide the variation of the critical radius for hot-jet ignition with both the jet velocity and the equivalence ratio of the mixture, giving values that vary between a few tens microns to a few hundred microns in the range of conditions explored. For a given equivalence ratio, the critical radius is found to increase with increasing injection velocities, although the increase is only moderately large. On the other hand, for a given injection velocity, the smallest critical radius is found at stoichiometric conditions.

Multi-distributed wireless sensors for monitoring a long distance transport in a reefer container

The study of the temperature gradients in cold stores and containers is a critical issue in the food industry for the quality assurance of products during transport and for minimising losses. This work presents an analysis of the temperatures during the refrigerated transport of 4,320 kg of blueberries in a reefer (set point temperature at ?1ºC) on a container ship from Montevideo (Uruguay) to Verona (Italy). The monitoring was performed by using semi-passive RFID loggers (TurboTag cards). The objective was to carry out a multi-distributed supervision using low-cost, wireless and autonomous sensors for the characterisation of the distribution and spatial gradients of temperatures during a long distance transport. Data analysis shows spatial (phase space) and temporal sequencing diagrams and reveals a significant heterogeneity of temperature at different locations in the container, which highlights the ineffectiveness of a temperature control system based on a single sensor, as is usually done.

Emergencia y convergencia de las corrientes urbanísticas alternativas al desarrollo disperso. Europa y los EE.UU. 1980/2010

La presente tesis doctoral identifica el reciente fenómeno de la emergenciai de corrientes urbanísticas que actúan en la periferia urbana a partir de una concepción del diseño urbano que propugna modelos más compactos, a escala humana, de carácter integral y sostenible, como alternativa al desarrollo disperso y difusoii que caracteriza en gran medida a la ciudad contemporánea. Se develan las diferencias de origen y las coincidencias entre las distintas líneas de pensamiento que definen a estas emergentes corrientes alternativas, situándolas dentro de determinados enfoques planteados en la presente tesis, y demostrando con ello, las progresivas áreas de convergencia que permiten su ordenamiento como fenómeno histórico común. Estas distintas líneas de pensamiento, que derivan en las corrientes urbanísticas alternativas, se manifiestan primeramente como reacción a las operaciones modernistas de la posguerra, pero crecientemente expresan una visión contestataria a la expansión urbana descontrolada iii, de bajas densidades, zonificación excluyente y dependiente del automóvil; modelo asociado a las autopistas urbanas, los llamados nuevos artefactos urbanos y las grandes parcelas destinadas a proyectos habitacionales. Una realidad expresada en variados contextos en el ámbito de la globalización económica y cultural que, en gran medida, se singulariza como responsable de las profundas transformaciones urbanas, de la consecuente gradual pérdida de habitabilidad en las ciudades y del impacto sobre el medio natural; cuestión a tratar en el “Marco conceptual” referido al contexto y problema de la tesis. Se estudian los planteamientos fundamentales que postulan estas corrientes alternativas en el contexto de la expansión urbana horizontal, con el fin de identificar las distintas vertientes o líneas existentes, de establecer sus orígenes, enfoques, vínculos y alcances, permitiendo con ello caracterizar el fenómeno en sí desde una visión original e integradora. Estas corrientes a tratar han surgido en diversos momentos y con diversas particularidades, pero con mayor intensidad, articulación y visibilidad a partir de los años ochenta y especialmente en los noventa del siglo recién pasado, fundamentalmente en Europa y en los Estados Unidos de América, representando el “Cuerpo teórico” de esta tesis. Emergen en un escenario de creciente empoderamiento ciudadano, en tiempos de una mayor conciencia ambiental y social, y desde la convicción extendida de las limitaciones de un desarrollismo que, en términos del territorio físico, se manifiesta en el crecimiento urbano por extensión, incremental y desarticulado; tienen distintos idearios de origen, algunos en aparente contradicción o manifiesta oposición entre sí; se articulan desde diversos énfasis y surgen a partir de distintas aproximaciones. Para poder comprender el fenómeno de manera integral, se identifican y observan las tendencias contemporáneas y las correspondientes corrientes principales del diseño urbano; se indaga en sus discursos críticos y se ordenan en función de tres enfoques, los cuales reúnen los fundamentos esenciales de las distintas miradas críticas al modelo disperso y difuso: el enfoque neotradicional, el enfoque de la movilidad, y el enfoque ecológicoiv. Reconociendo la importancia de las operaciones de renovación urbana en la “ciudad intramuros” y siendo partícipes de estas intervenciones, las diferentes corrientes observadas desde los tres enfoques planteados, tienen un común denominador disciplinar en cuanto a su preocupación por generar actuaciones urbanas viables y practicables que causen un menor impacto sobre el territorio y que ofrezcan las condiciones para mejorar la habitabilidad en la ciudad, asumiendo el crecimiento hacia las periferias desde una aproximación alternativa a los modelos hoy predominantes. Las distintas corrientes, desde los particulares enfoques señalados, pretenden en síntesis, un desarrollo que refuerce el rol del diseño urbano, que promueva modelos más compactosv, de usos mixtos compatibles con la residencia y con espacios públicos de calidad, a una escala apropiada para la movilidad peatonal, favoreciendo de esta manera el uso del transporte público y velando consecuentemente por aquellos aspectos que mejoren las condiciones de sostenibilidad. En definitiva, el ideario de las corrientes caracterizadas en esta tesis como alternativas, se origina y se sostiene a partir de la convicción de situar a la disciplina del diseño urbano en su justa medida en el desarrollo contemporáneo, en el convencimiento de la relevancia del proyecto urbano en el ordenamiento del territorio y el crecimiento de las ciudades actuales. Una convergencia implícita a tratarse en la “Discusión de resultados”. La preocupación que reflejan las distintas corrientes estudiadas por el hábitat construido, en cuanto a su condición de acoger y estimular la dimensión humana de las ciudades, puede resumirse en una cita del arquitecto danés Jan Gehl, que interpreta a muchas de las líneas de pensamiento que se abordan en esta tesis, en cuanto al deseo de crear mejores ciudades: “Aspiramos generar ciudades animadas, saludables, atractivas, sostenibles y seguras, con el propósito de mejorar la calidad de vida de la gente”. ABSTRACT This Ph.D thesis identifies the recent phenomenon of the emergence of alternative urban design currents in the city edge, that focus on a more compact, human-scale oriented, comprehensive and sustainable urban design as an alternative to the overwhelming advance of sprawl that largely characterizes the contemporary city. It singles out the differences in origin, as well as the similarities drawn from the different lines of thought that define these emergent alternative currents, placing them within certain approaches proposed in this thesis, and thereby demonstrating the progressive areas of convergence that allow to arrange them as a common historical phenomenon. Indeed, the different lines of thought that drift towards these alternative urbanistic currents, first appear as a reaction to the post- World War II Modernist housing projects, and increasingly more so as a response to urban sprawl; an ever expanding phenomemnon expressed in different contexts within the economic and cultural globalization, largely singled out as being responsible for the deep urban transformations, the consequent gradual loss of livability and the impact on natural environment, setting the “Conceptual framework” of this thesis. Within the context of urban sprawl, the fundamental principles of these alternative currents are studied to establish their origins, approaches, linkages and scope, in order to zero in on the various aspects or existing lines and thereby allowing the identification of the phenomenon from an original and inclusive vision. The alternative currents to be dealt with, emerged at different times and with diverse characteristics, nonetheless it is in the eighties and especially during the nineties, mainly in Europe and the United States, where they appear with greater intensity, coordination and visibility, representing the “Theoretical body” of this thesis. The above mentioned currents arise in a scenario of increasing citizen empowerment in times of growing environmental and social awareness, along with the widespread conviction of the limitations of a quantitative development expressed as a physical manifestation in urban sprawl. These alternative currents have a different philosophy in origin, some are in apparent contradiction or in definite opposition, being organized from different approaches and emphases. To comprehend this phenomenon, it is necessary to identify and analize the contemporary trends and the main currents of urban design, inquiring and arranging their critical discourses according to three approaches that meet the essential foundations of the various critical perspectives of urban sprawl: the neo-traditional approach, the mobility approach, the ecological approach. Acknowledging the importance of urban renovation in the “Inner City”, and being partakers of these interventions, the different currents identified from these three approaches, concur in their concern for the development of viable and workable alternatives to urban sprawl; models that cause a lower impact on the territory, whilst improving livability in the city. The different currents recognized within the above mentioned approaches, advocate in brief, for a development that strengthens the role of urban design, that encourages more compact models, mixed-uses, and quality public spaces, considering the human scale and walkability; thereby increasing public transport and promoting consequently, the conditions for sustainable development. Ultimately, the philosophy of the so called alternative currents, is based on the belief of placing urban design as a fundamental discipline in contemporary urban development. The concern for the built habitat reflected by the various currents studied, as of their condition to accept and stimulate the human dimension of the city, can be summed up in a quote from Danish architect Jan Gehl, who represents many of the lines of thought addressed, in the belief of creating better cities: “We aspire to create cities that are lively, healthy, attractive, sustainable and safe-and thereby improve people’s quality of life”.

Conformational changes couple Na+ and glucose transport

The mechanism by which cotransport proteins couple their substrates across cell membranes is not known. A commonly proposed model is that cotransport results from ligand-induced conformational transitions that change the accessibility of ligand-binding sites from one side of the membrane to the other. To test this model, we have measured the accessibility of covalent probes to a cysteine residue (Q457C) placed in the putative sugar-translocation domain of the Na+/glucose cotransporter (SGLT1). The mutant protein Q457C was able to transport sugar, but transport was abolished after alkylation by methanethiosulfonate reagents. Alkylation blocked sugar translocation but not sugar binding. Accessibility of Q457C to alkylating reagents required external Na+ and was blocked by external sugar and phlorizin. The voltage dependence of accessibility was directly correlated with the presteady–state charge movement of SGLT1. Voltage-jump experiments with rhodamine-6-maleimide-labeled Q457C showed that the time course and level of changes in fluorescence closely followed the presteady–state charge movement. We conclude that conformational changes are responsible for the coupling of Na+ and sugar transport and that Q457 plays a critical role in sugar translocation by SGLT1.

The maturity-onset diabetes of the young (MODY1) transcription factor HNF4α regulates expression of genes required for glucose transport and metabolism

Hepatocyte nuclear factor 4α (HNF4α) plays a critical role in regulating the expression of many genes essential for normal functioning of liver, gut, kidney, and pancreatic islets. A nonsense mutation (Q268X) in exon 7 of the HNF4α gene is responsible for an autosomal dominant, early-onset form of non-insulin-dependent diabetes mellitus (maturity-onset diabetes of the young; gene named MODY1). Although this mutation is predicted to delete 187 C-terminal amino acids of the HNF4α protein the molecular mechanism by which it causes diabetes is unknown. To address this, we first studied the functional properties of the MODY1 mutant protein. We show that it has lost its transcriptional transactivation activity, fails to dimerize and bind DNA, implying that the MODY1 phenotype is because of a loss of HNF4α function. The effect of loss of function on HNF4α target gene expression was investigated further in embryonic stem cells, which are amenable to genetic manipulation and can be induced to form visceral endoderm. Because the visceral endoderm shares many properties with the liver and pancreatic β-cells, including expression of genes for glucose transport and metabolism, it offers an ideal system to investigate HNF4-dependent gene regulation in glucose homeostasis. By exploiting this system we have identified several genes encoding components of the glucose-dependent insulin secretion pathway whose expression is dependent upon HNF4α. These include glucose transporter 2, and the glycolytic enzymes aldolase B and glyceraldehyde-3-phosphate dehydrogenase, and liver pyruvate kinase. In addition we have found that expression of the fatty acid binding proteins and cellular retinol binding protein also are down-regulated in the absence of HNF4α. These data provide direct evidence that HNF4α is critical for regulating glucose transport and glycolysis and in doing so is crucial for maintaining glucose homeostasis.

Glutamate transporter currents in Bergmann glial cells follow the time course of extrasynaptic glutamate

Glutamate transporters in the central nervous system are expressed in both neurons and glia, they mediate high affinity, electrogenic uptake of glutamate, and they are associated with an anion conductance that is stoichiometrically uncoupled from glutamate flux. Although a complete cycle of transport may require 50–100 ms, previous studies suggest that transporters can alter synaptic currents on a much faster time scale. We find that application of l-glutamate to outside-out patches from cerebellar Bergmann glia activates anion-potentiated glutamate transporter currents that activate in <1 ms, suggesting an efficient mechanism for the capture of extrasynaptic glutamate. Stimulation in the granule cell layer in cerebellar slices elicits all or none α-amino-3-hydroxy-5-methyl-4-isoxazolepropionate receptor and glutamate transporter currents in Bergmann glia that have a rapid onset, suggesting that glutamate released from climbing fiber terminals escapes synaptic clefts and reaches glial membranes shortly after release. Comparison of the concentration dependence of both α-amino-3-hydroxy-5-methyl-4-isoxazolepropionate receptor and glutamate transporter kinetics in patches with the time course of climbing fiber-evoked responses indicates that the glutamate transient at Bergmann glial membranes reaches a lower concentration than attained in the synaptic cleft and remains elevated in the extrasynaptic space for many milliseconds.

The Nucleoporin Nup153 Plays a Critical Role in Multiple Types of Nuclear Export

The fundamental process of nucleocytoplasmic transport takes place through the nuclear pore. Peripheral pore structures are presumably poised to interact with transport receptors and their cargo as these receptor complexes first encounter the pore. One such peripheral structure likely to play an important role in nuclear export is the basket structure located on the nuclear side of the pore. At present, Nup153 is the only nucleoporin known to localize to the surface of this basket, suggesting that Nup153 is potentially one of the first pore components an RNA or protein encounters during export. In this study, anti-Nup153 antibodies were used to probe the role of Nup153 in nuclear export in Xenopus oocytes. We found that Nup153 antibodies block three major classes of RNA export, that of snRNA, mRNA, and 5S rRNA. Nup153 antibodies also block the NES protein export pathway, specifically the export of the HIV Rev protein, as well as Rev-dependent RNA export. Not all export was blocked; Nup153 antibodies did not impede the export of tRNA or the recycling of importin β to the cytoplasm. The specific antibodies used here also did not affect nuclear import, whether mediated by importin α/β or by transportin. Overall, the results indicate that Nup153 is crucial to multiple classes of RNA and protein export, being involved at a vital juncture point in their export pathways. This juncture point appears to be one that is bypassed by tRNA during its export. We asked whether a physical interaction between RNA and Nup153 could be observed, using homoribopolymers as sequence-independent probes for interaction. Nup153, unlike four other nucleoporins including Nup98, associated strongly with poly(G) and significantly with poly(U). Thus, Nup153 is unique among the nucleoporins tested in its ability to interact with RNA and must do so either directly or indirectly through an adaptor protein. These results suggest a unique mechanistic role for Nup153 in the export of multiple cargos.

Rab11 Is Required for Trans-Golgi Network–to–Plasma Membrane Transport and a Preferential Target for GDP Dissociation Inhibitor

The rab11 GTPase has been localized to both the Golgi and recycling endosomes; however, its Golgi-associated function has remained obscure. In this study, rab11 function in exocytic transport was analyzed by using two independent means to perturb its activity. First, expression of the dominant interfering rab11S25N mutant protein led to a significant inhibition of the cell surface transport of vesicular stomatitis virus (VSV) G protein and caused VSV G protein to accumulate in the Golgi. On the other hand, the expression of wild-type rab11 or the activating rab11Q70L mutant had no adverse effect on VSV G transport. Next, the membrane association of rab11, which is crucial for its function, was perturbed by modest increases in GDP dissociation inhibitor (GDI) levels. This led to selective inhibition of the trans-Golgi network to cell surface delivery, whereas endoplasmic reticulum–to–Golgi and intra-Golgi transport were largely unaffected. The transport inhibition was reversed specifically by coexpression of wild-type rab11 with GDI. Under the same conditions two other exocytic rab proteins, rab2 and rab8, remained membrane bound, and the transport steps regulated by these rab proteins were unaffected. Neither mutant rab11S25N nor GDI overexpression had any impact on the cell surface delivery of influenza hemagglutinin. These data show that functional rab11 is critical for the export of a basolateral marker but not an apical marker from the trans-Golgi network and pinpoint rab11 as a sensitive target for inhibition by excess GDI.