Roles of Predicted Glycosyltransferases in the Biosynthesis of the Rhizobium etli CE3 O Antigen

The Rhizobium etli CE3 O antigen is a fixed-length heteropolymer. The genetic regions required for its synthesis have been identified, and the nucleotide sequences are known. The structure of the O antigen has been determined, but the roles of specific genes in synthesizing this structure are relatively unclear. Within the known O-antigen genetic clusters of this strain, nine open reading frames (ORFs) were found to contain a conserved glycosyltransferase domain. Each ORF was mutated, and the resulting mutant lipopolysaccharide (LPS) was analyzed. Tricine SDS-PAGE revealed stepwise truncations of the O antigen that were consistent with differences in mutant LPS sugar compositions and reactivity with O-antigen-specific monoclonal antibodies. Based on these results and current theories of O-antigen synthesis, specific roles were deduced for each of the nine glycosyltransferases, and a model for biosynthesis of the R. etli CE3 O antigen was proposed. In this model, O-antigen biosynthesis is initiated with the addition of N-acetyl-quinovosamine-phosphate (QuiNAc-P) to bactoprenol-phosphate by glycosyltransferase WreU. Glycosyltransferases WreG, WreE, WreS, and WreT would each act once to attach mannose, fucose, a second fucose, and 3-O-methyl-6-deoxytalose (3OMe6dTal), respectively. WreH would then catalyze the addition of methyl glucuronate (MeGlcA) to complete the first instance of the O-antigen repeat unit. Four subsequent repeats of this unit composed of fucose, 3OMe6dTal, and MeGlcA would be assembled by a cycle of reactions catalyzed by two additional glycosyltransferases, WreM and WreL, along with WreH. Finally, the O antigen would be capped by attachment of di- or tri-O-methylated fucose as catalyzed by glycosyltransferase WreB

Genetic Basis for \u3cem\u3eRhizobium etli\u3c/em\u3e CE3 O-Antigen O-Methylated Residues That Vary According to Growth Conditions

The Rhizobium etli CE3 O antigen is a fixed-length heteropolymer with O methylation being the predominant type of sugar modification. There are two O-methylated residues that occur, on average, once per complete O antigen: a multiply O-methylated terminal fucose and 2-O methylation of a fucose residue within a repeating unit. The amount of the methylated terminal fucose decreases and the amount of 2-O-methylfucose increases when bacteria are grown in the presence of the host plant, Phaseolus vulgaris, or its seed exudates. Insertion mutagenesis was used to identify open reading frames required for the presence of these O-methylated residues. The presence of the methylated terminal fucose required genes wreA, wreB, wreC, wreD, and wreF, whereas 2-O methylation of internal fucoses required the methyltransferase domain of bifunctional gene wreM. Mutants lacking only the methylated terminal fucose, lacking only 2-O methylation, or lacking both the methylated terminal fucose and 2-O methylation exhibited no other lipopolysaccharide structural defects. Thus, neither of these decorations is required for normal O-antigen length, transport, or assembly into the final lipopolysaccharide. This is in contrast to certain enteric bacteria in which the absence of a terminal decoration severely affects O-antigen length and transport. R. etli mutants lacking only the methylated terminal fucose were not altered in symbiosis with host Phaseolus vulgaris, whereas mutants lacking only 2-O-methylfucose exhibited a delay in nodule development during symbiosis. These results support previous conclusions that the methylated terminal fucose is dispensable for symbiosis, whereas 2-O methylation of internal fucoses somehow facilitates early events in symbiosis

Centaur AC-8 Postflight Guidance Analysis

Centaur AC-8 postflight guidance and control analysi

La población de España en el siglo XVII. Tratamiento demográfico de la bula de la Santa Cruzada

El artículo reflexiona entorno las dificultades metodológicas de estudiar el siglo XVII desde el punto de vista demográfico a partir del análisis de la Bula de la Santa Cruzada.A l'article es reflexiona entorn les dificultats metodològiques d'estudiar el segle XVII des del punt de vista demogràfic a partir de l'anàlisi de la Butlla de la Santa Cruzada.The article focus on the metodological difficulties to study the 17th century. On a demographic point of view with an analysis of the Bull of the Saint Crusade

SDR: Las radios definidas por software requieren pruebas avanzadas

Hoy en día y más que antes, las tecnologías de radio frecuencia (RF) se están desarrollando para su adaptación - adaptación a un espectro de RF muy poblado - a los diversos requisitos de la red y a los cambios en las especificaciones de los dispositivos. En ningún otro lugar es esto más evidente que en el caso de SDR (Software Defined Radios) que permiten a un software controlar de forma dinámica los parámetros de las comunicaciones tales como la banda de frecuencias utilizada, el tipo de modulación, los tipos de datos y los esquemas de saltos de frecuencia

Wear and friction of TiAlN/VN coatings against Al2O3 in air at room and elevated temperatures

TiAlN/VN multilayer coatings exhibit excellent dry sliding wear resistance and low friction coefficient, reported to be associated with the formation of self-lubricating V2O5. To investigate this hypothesis, dry sliding ball-on-disc wear tests of TiAlN/VN coatings on flat stainless steel substrates were undertaken against Al2O3 at 25 C, 300 C and 635 C in air. The coating exhibited increased wear rate with temperature. The friction coefficient was 0.53 at 25 C, which increased to 1.03 at 300 C and decreased to 0.46 at 635 C. Detailed investigation of the worn surfaces was undertaken using site-specific transmission electron microscopy (TEM) via focused ion beam (FIB) microscopy, along with Fourier transform infrared (FTIR) and Raman spectroscopy. Microstructure and tribo-induced chemical reactions at these temperatures were correlated with the coating’s wear and friction behaviour. The friction behaviour at room temperature is attributed to the presence of a thin hydrated tribofilm and the presence of V2O5 at high temperature

Affine semigroups having a unique Betti element

We characterize affine semigroups having one Betti element and we compute some relevant non-unique factorization invariants for these semigroups. As an example, we particularize our description to numerical semigroups.Comment: 8 pages, 1 figure. To appear in Journal of Algebra and its Application

Do indigenous peoples benefit from poverty programs? Evidence from Mexico´s 2000 census

Indigenous peoples are among the poorest in Latin America, and it is often argued that social policies do not reach them. At the same time, several countries have implemented in recent years new programs for poverty reduction that should have benefited the indigenous. In this paper, we use data from Mexico's 2000 census to test whether indigenous peoples living in the southern states of Chiapas, Guerrero and Oaxaca benefit from three large government programs: PROGRESA, FISM, and PROCAMPO.