Functional invalidation of the autotaxin gene by a single amino acid mutation in mouse is lethal

AbstractAutotaxin is a member of the phosphodiesterase family of enzymes, (NPP2). It is an important secreted protein found in conditioned medium from adipocytes. It also has a putative role in the metastatic process. Based on these observation, further validation of this potential target was necessary, apart from the classical biochemical ones. The construction of a knock out mouse strain for ATX was started. In this paper, we report the generation of a mouse line displaying an inactivated ATX gene product. The KO line was designed in order to generate a functional inactivation of the protein. In this respect, the threonine residue T210 was replaced by an alanine (T210A) leading to a catalytically inactive enzyme. If the experimental work was straight forward, we disappointedly discovered at the final stage that the breeding of heterozygous animals, ATX −/+, led to the generation of a Mendelian repartition of wild-type and heterozygous, but no homozygous were found, strongly suggesting that the ATX deletion is lethal at an early stage of the development. This was confirmed by statistical analysis. Although other reported the same lethality for attempted ATX−/− mice generation [van Meeteren, L.A., Ruurs, P., Stortelers, C., Bouwman, P., van Rooijen, M.A., Pradère, J.P., Pettit, T.R., Wakelam, M.J.O., Saulnier-Blache, J.S., Mummery, C.L., Moolenar, W.H. and Jonkers, J. (2006) Autotaxin, a secreted lysophospholipase D, is essential for blood vessel formation during development, Mol. Cell. Biol. 26, 5015–5022; Tanaka, M., Okudaira, S., Kishi, Y., Ohkawa, R., Isei, S., Ota, M., Noji, S., Yatomi, Y., Aoki, J., and Arai, H. (2006) Autotaxin stabilizes blood vessels and is required for embryonic vasculature by producing lysophosphatidic acid, J. Biol. Chem. 281, 25822–25830], they used more drastic multiple exon deletions in the ATX gene, while we chose a single point mutation. To our knowledge, the present work is the first showing such a lethality in any gene after a point mutation in an enzyme catalytic site

A multi-site campaign to measure solar-like oscillations in Procyon. II. Mode frequencies

We have analyzed data from a multi-site campaign to observe oscillations in the F5 star Procyon. The data consist of high-precision velocities that we obtained over more than three weeks with eleven telescopes. A new method for adjusting the data weights allows us to suppress the sidelobes in the power spectrum. Stacking the power spectrum in a so-called echelle diagram reveals two clear ridges that we identify with even and odd values of the angular degree (l=0 and 2, and l=1 and 3, respectively). We interpret a strong, narrow peak at 446 muHz that lies close to the l=1 ridge as a mode with mixed character. We show that the frequencies of the ridge centroids and their separations are useful diagnostics for asteroseismology. In particular, variations in the large separation appear to indicate a glitch in the sound-speed profile at an acoustic depth of about 1000 s. We list frequencies for 55 modes extracted from the data spanning 20 radial orders, a range comparable to the best solar data, which will provide valuable constraints for theoretical models. A preliminary comparison with published models shows that the offset between observed and calculated frequencies for the radial modes is very different for Procyon than for the Sun and other cool stars. We find the mean lifetime of the modes in Procyon to be 1.29 +0.55/-0.49 days, which is significantly shorter than the 2-4 days seen in the Sun.Comment: accepted for publication in Ap

Receptor Complementation and Mutagenesis Reveal SR-BI as an Essential HCV Entry Factor and Functionally Imply Its Intra- and Extra-Cellular Domains

HCV entry into cells is a multi-step and slow process. It is believed that the initial capture of HCV particles by glycosaminoglycans and/or lipoprotein receptors is followed by coordinated interactions with the scavenger receptor class B type I (SR-BI), a major receptor of high-density lipoprotein (HDL), the CD81 tetraspanin, and the tight junction protein Claudin-1, ultimately leading to uptake and cellular penetration of HCV via low-pH endosomes. Several reports have indicated that HDL promotes HCV entry through interaction with SR-BI. This pathway remains largely elusive, although it was shown that HDL neither associates with HCV particles nor modulates HCV binding to SR-BI. In contrast to CD81 and Claudin-1, the importance of SR-BI has only been addressed indirectly because of lack of cells in which functional complementation assays with mutant receptors could be performed. Here we identified for the first time two cell types that supported HCVpp and HCVcc entry upon ectopic SR-BI expression. Remarkably, the undetectable expression of SR-BI in rat hepatoma cells allowed unambiguous investigation of human SR-BI functions during HCV entry. By expressing different SR-BI mutants in either cell line, our results revealed features of SR-BI intracellular domains that influence HCV infectivity without affecting receptor binding and stimulation of HCV entry induced by HDL/SR-BI interaction. Conversely, we identified positions of SR-BI ectodomain that, by altering HCV binding, inhibit entry. Finally, we characterized alternative ectodomain determinants that, by reducing SR-BI cholesterol uptake and efflux functions, abolish HDL-mediated infection-enhancement. Altogether, we demonstrate that SR-BI is an essential HCV entry factor. Moreover, our results highlight specific SR-BI determinants required during HCV entry and physiological lipid transfer functions hijacked by HCV to favor infection

Mémoires interrompus

Un germaniste hors du commun nous livre ici ses souvenirs et nous fait bénéficier des leçons qu'il a tirées de la fréquentation des puissants comme des humbles, des salons comme des prisons, des palais de la République comme des paysages sauvages de ses Pyrénées tant aimées.An outstanding German studies specialist relates here his memories. He transmits to us the life lessons he derived from the company of the powerful and the humble, in both lounges and prisons, in the Republique’s Palaces as in the wild landscapes of his much loved Pyrenees

Functional role of TTF-1 binding sites in bovine thyroglobulin promoter

We have studied the binding of purified TTF-1 on the bovine thyroglobulin gene promoter. DNase I footprinting experiments revealed three binding sites which corresponded in location to the A, B and C sites found in the rat thyroglobulin promoter. Mutants in the A and C regions showing reduced binding of TTF-1, also exhibited largely decreased promoter activity in transient expression experiments in primary-cultured dog thyrocytes. Two mutants in the B site that exhibited a reduced capacity to bind TTF-1 also displayed a drastically affected transcriptional activity in transient assays. As in the rat, sites A and C only are critical for promoter activity, these results suggest that full occupancy of the B site is required for thyroglobulin promoter activity in the cow only.Comparative StudyJournal ArticleResearch Support, Non-U.S. Gov'tinfo:eu-repo/semantics/publishe

Trajectoires familiales et espaces de vie en milieu urbain

L'étude des contextes d'habitat et des itinéraires résidentiels permet-elle de mieux comprendre ce qui se joue dans la vie du groupe domestique, dans les relations avec la parentèle et dans les rapports entre les générations ? Dans quelle mesure le logement et l'inscription dans des territoires sont-ils révélateurs de dynamiques familiales ? Réciproquement, quels effets ces dynamiques exercent-elles sur la localisation des ménages, sur la physionomie des quartiers, sur la vie des différents lieux de la ville ? Comment les mobilités choisies, subies ou projetées par les familles interfèrent-elles avec le jeu du marché immobilier et la mise en œuvre des politiques urbaines ? En quoi l'analyse des trajectoires familiales enrichit-elle notre connaissance des rapports que les citadins entretiennent avec leur habitat, des usages qu'ils font de la ville, et des modalités selon lesquelles s'organise leur coexistence avec d'autres citadins ? Les textes rassemblés dans ce volume se situent au croisement de ces différentes questions. Ils émanent de sociologues, de démographes, d'historiens et de géographes qui avaient été invités à confronter leurs travaux à l'occasion d'un colloque international organisé par le Centre Jacques Cartier

Recherche en sciences humaines sur l'Asie du Sud-Est

Moussons porte son regard sur le Sud-Est asiatique (insulaire et continental) et ses marges indiennes et chinoises. Revue comparative et interdisciplinaire, elle accueille l'ensemble des sciences humaines et sociales, plus particulièrement l’anthropologie, la sociologie et l’histoire contemporaine. Revue biannuelle, Moussons publie, en français et en anglais, des articles, des notes de recherche et des comptes rendus d’ouvrages

A Multisite Campaign to Measure Solar-Like Oscillations in Procyon. I. Observations, Data Reduction and Slow Variations

International audienceWe have carried out a multisite campaign to measure oscillations in the F5 star Procyon A. We obtained high-precision velocity observations over more than three weeks with 11 telescopes, with almost continuous coverage for the central 10 days. This represents the most extensive campaign so far organized on any solar-type oscillator. We describe in detail the methods we used for processing and combining the data. These involved calculating weights for the velocity time series from the measurement uncertainties and adjusting them in order to minimize the noise level of the combined data. The time series of velocities for Procyon shows the clear signature of oscillations, with a plateau of excess power that is centered at 0.9 mHz and is broader than has been seen for other stars. The mean amplitude of the radial modes is 38:1 AE 1:3 cm s À1 (2.0 times solar), which is consistent with previous detections from the ground and by the WIRE spacecraft, and also with the upper limit set by the MOST spacecraft. The variation of the amplitude during the observing campaign allows us to estimate the mode lifetime to be 1:5 þ1:9 À0:8 days. We also find a slow variation in the radial velocity of Procyon, with good agreement between different telescopes. These variations are remarkably similar to those seen in the Sun, and we interpret them as being due to rotational modulation from active regions on the stellar surface. The variations appear to have a period of about 10 days, which presumably equals the stellar rotation period or, perhaps, half of it. The amount of power in these slow variations indicates that the fractional area of Procyon covered by active regions is slightly higher than for the Sun