Multifunctional nature of P fimbriae of uropathogenic Escherichia coli: mutations in fsoE and feoF influence fimbrial binding to renal tubuli and immobilized fibronectin

P fimbriae of the F7(1) serotype of Escherichia coli are composed of a major subunit, FsoA, and of three minor proteins named FsoG, FsoE, and FsoF. FsoG is the Gal alpha(1-4)Gal-specific lectin. We assessed mutated recombinant strains each deficient in one fimbrial component for adhesion to frozen sections of rat cortical kidney and to fibronectin immobilized on glass. Rat kidney lacks the Gal alpha(1-4)Gal-containing glycolipids. The fsoG mutant strain was as adhesive to sections of rat kidney and to fibronectin-coated glass as was the recombinant strain expressing the complete fso gene cluster. The fsoA mutant strain was highly adhesive to fibronectin and to kidney sections. In the rat kidney, the adhesion of these strains was predominantly localized to sites of basolateral membranes of tubuli. The fsoE and the fsoF mutant strains were slightly less adhesive to kidney structures and failed to adhere to fibronectin. The fsoE, fsoF double mutant strain adhered neither to fibronectin nor to kidney sections. None of the fso recombinant strains reacted with soluble fibronectin, suggesting that the interaction is dependent on the conformation of the fibronectin molecules. Recombinant strains expressing the F7(2), F8, F11, F13, and F14 serovariants of the P fimbria also showed adherence to immobilized fibronectin. The results show that in addition to binding to globoseries of glycolipids via the G protein, the P fimbriae of uropathogenic E. coli exhibit a tissue-binding property influenced by fsoE and fsoF gene products and with affinity for basolateral membranes and fibronectin.     

Lectins facilitate calcium-induced fusion of phospholipid vesicles containing glycosphingolipids

Ca2+-induced fusion of phospholipid vesicles containing globoside (GL-4) or disialoganglioside (GDla) is several-fold slower than the fusion of the pure phospholipid vesicles. Lectins specific for these glycosphingolipids, soybean agglutinin and wheat germ agglutinin, respectively, enhance the rate of fusion when added to the vesicle suspension before the introduction of Ca2+. The enhancement depends on the lectin concentration and the time of preincubation with the lectin. We propose that lectins facilitate membrane fusion by inducing intermembrane contact, which is the first step in the overall process of membrane fusion, or by laterally phase separating the inhibitory glycolipids.     

Accounting for differential migration strategies between age groups to monitor raptor population dynamics in the eastern Black Sea flyway

Migration counts can offer a cost-effective method for monitoring the state of migrant raptor populations. However, differential migration strategies between inexperienced juveniles and experienced non-juveniles are rarely accounted for when inferring population trends from raptor migration counts. Since 2011, the Batumi Raptor Count (BRC) monitors the autumn migration of more than 1 million raptors along the eastern Black Sea coast in the Republic of Georgia. We also systematically sampled age information to assess differential migration timing between age groups and estimate age-specific linear trends in abundance between 2011 and 2018 for eight focal species. In so doing we aimed (1) to reassess the global relevance of BRC counts for each species and the potential for monitoring abundance of juveniles and non-juveniles, and (2) to identify demographic changes underlying recent trends in overall abundance. We found that the mean annual passage of non-juveniles at Batumi represents at least 1% of the estimated global breeding population of five study species. As expected, counts of juveniles were more variable than counts of non-juveniles. Yet despite our short monitoring period our models had sufficient statistical power to detect changes in abundance of 10%/year or less for at least one age group in all species except Pallid Harrier Circus macrourus. Our results indicate stable abundance and demography for half of the study species. We also found strong and significant increases in the abundance of Black Kites Milvis migrans and Short-toed Eagles Circaetus gallicus that were primarily due to increasing numbers of non-juveniles. By contrast, juvenile Montagu's Harriers Circus pygargus and Booted Eagles Hieraaetus pennatus significantly decreased in abundance. The first decade of BRC surveys offers an important benchmark for monitoring raptor populations using the eastern African–Palearctic flyway in the 21st century. We discuss possible causes of the observed trends and hope our work will stimulate demographic monitoring at migration count sites.     

Ueber Virulenz und Virulenz-Curven

Zusammenfassung Es wird eine Methode angegeben, um die Virulenz graphisch darzustellen. Der Verlauf der Curve wird durch Aenderung der Factoren sowohl des Erregers als auch des Wirtes deutlich beeinflusst.     

Protein kinase C prevents oligodendrocyte differentiation: modulation of actin cytoskeleton and cognate polarized membrane traffic.

In a previous study, we showed that activation of protein kinase C (PKC) prevents oligodendrocyte differentiation at the pro-oligodendrocyte stage. The present study was undertaken to identify downstream targets of PKC action in oligodendrocyte progenitor cells. Activation of PKC induced the predominant phosphorylation of an 80-kD protein, identified as myristoylated alanine-rich C-kinase substrate (MARCKS). Upon phosphorylation, MARCKS is translocated from the plasma membrane to the cytosol. Furthermore, PKC activation perturbed the organization of the actin cytoskeleton, causing a redistribution of actin filaments to the submembranous or cortical actin cytoskeleton. As a consequence, transport of a protein traffic marker, the vesicular stomatitis virus glycoprotein, from the trans-Golgi network to the plasma membrane becomes perturbed. The effect of disruption of the actin filament network by cytochalasin D perfectly matched the effect of PKC. These data thus favor the existence of a causal relationship between actin rearrangement and docking and/or fusion of proteins to the plasma membrane. Interestingly, neither in control cells nor in PKC-activated cells did another protein traffic marker, influenza hemagglutinin (HA), reach the cell surface. However, an eminent and specific accumulation of HA just underneath the plasma membrane became apparent upon PKC activation. Yet, this effect could not be simulated by cytochalasin D treatment. Therefore, these observations imply that although MARCKS represents a prominent PKC target site in regulating differentiation, another target involves the differential control of cognate polarized trafficking pathways, which are apparently operating in oligodendrocyte progenitor cells.     

Relationship between synthesis of ribosomes and RNA polymerase in Bacillus subtilis

The relationship between RNA polymerase synthesis and ribosome synthesis has been investigated in Bacillus subtilis growing at different rates. The amount of polymerase was determined by quantitation of polymerase subunits on polyacrylamide gels, while ribosome synthesis was estimated from accumulation of total RNA. The ratio of ribosome to RNA polymerase synthesis was 5:1 at slower growth rates and increased with growth rate. Bacillus subtilis was estimated to contain between 10-and 2-fold excess of RNA polymerase depending on its growth rate.     

Cloning of an exclusion-determining fragment of the IncI plasmid, R144

By cloning a distinct 8 MDa fragment of the IncI plasmid, R144, in the vector pACYC184, two recombinant plasmids were isolated. In these plasmids, pRAH303 and pRAH308, the inserted fragment was in opposite orientations. Both plasmids when present in a recipient strain caused a conjugation-specific exclusion in crosses with donor cells carrying the IncI plasmid R144. Some derivatives of the recombinant plasmids in which parts were deleted, or in which Tn5 transposons were inserted, appeared to be exclusion negative. Analysis in minicells of the gene products of such plasmids together with those of the original recombinant plasmids revealed that the presence of two proteins, with apparent molecular weights of 13,000 and 19,000 Da could be correlated with the exclusion phenomenon.