Lamellipodium extension and membrane ruffling require different SNARE-mediated trafficking pathways

<p>Abstract</p> <p>Background</p> <p>Intracellular membrane traffic is an essential component of the membrane remodeling that supports lamellipodium extension during cell adhesion. The membrane trafficking pathways that contribute to cell adhesion have not been fully elucidated, but recent studies have implicated SNARE proteins. Here, the functions of several SNAREs (SNAP23, VAMP3, VAMP4 and syntaxin13) are characterized during the processes of cell spreading and membrane ruffling.</p> <p>Results</p> <p>We report the first description of a SNARE complex, containing SNAP23, syntaxin13 and cellubrevin/VAMP3, that is induced by cell adhesion to an extracellular matrix. Impairing the function of the SNAREs in the complex using inhibitory SNARE domains disrupted the recycling endosome, impeded delivery of integrins to the cell surface, and reduced haptotactic cell migration and spreading. Blocking SNAP23 also inhibited the formation of PMA-stimulated, F-actin-rich membrane ruffles; however, membrane ruffle formation was not significantly altered by inhibition of VAMP3 or syntaxin13. In contrast, membrane ruffling, and not cell spreading, was sensitive to inhibition of two SNAREs within the biosynthetic secretory pathway, GS15 and VAMP4. Consistent with this, formation of a complex containing VAMP4 and SNAP23 was enhanced by treatment of cells with PMA. The results reveal a requirement for the function of a SNAP23-syntaxin13-VAMP3 complex in the formation of lamellipodia during cell adhesion and of a VAMP4-SNAP23-containing complex during PMA-induced membrane ruffling.</p> <p>Conclusions</p> <p>Our findings suggest that different SNARE-mediated trafficking pathways support membrane remodeling during ECM-induced lamellipodium extension and PMA-induced ruffle formation, pointing to important mechanistic differences between these processes.</p

Absence of \u3ci\u3eStreptococcus pneumoniae \u3c/i\u3e Capsule Increases Bacterial Binding, Perisstence, and Inflammation In Corneal Infection

The role of the pneumococcal polysaccharide capsule is largely unclear for Streptococcus pneumoniae keratitis, an ocular inflammatory disease that develops as a result of bacterial infection of the cornea. In this study, capsule-deficient strains were compared to isogenic parent strains in their ability to adhere to human corneal epithelial cells. One isogenic pair was further used in topical ocular infection of mice to assess the contribution of the capsule to keratitis. The results showed that non-encapsulated pneumococci were significantly more adherent to cells, persisted in significantly higher numbers on mouse corneas in vivo, and caused significant increases in murine ocular IL9, IL10, IL12-p70, MIG, and MIP-1-gamma compared to encapsulated S. pneumoniae. These findings indicate that the bacterial capsule impedes virulence and the absence of capsule impacts inflammation following corneal infection

A Model-Based Analysis of GC-Biased Gene Conversion in the Human and Chimpanzee Genomes

GC-biased gene conversion (gBGC) is a recombination-associated process that favors the fixation of G/C alleles over A/T alleles. In mammals, gBGC is hypothesized to contribute to variation in GC content, rapidly evolving sequences, and the fixation of deleterious mutations, but its prevalence and general functional consequences remain poorly understood. gBGC is difficult to incorporate into models of molecular evolution and so far has primarily been studied using summary statistics from genomic comparisons. Here, we introduce a new probabilistic model that captures the joint effects of natural selection and gBGC on nucleotide substitution patterns, while allowing for correlations along the genome in these effects. We implemented our model in a computer program, called phastBias, that can accurately detect gBGC tracts about 1 kilobase or longer in simulated sequence alignments. When applied to real primate genome sequences, phastBias predicts gBGC tracts that cover roughly 0.3% of the human and chimpanzee genomes and account for 1.2% of human-chimpanzee nucleotide differences. These tracts fall in clusters, particularly in subtelomeric regions; they are enriched for recombination hotspots and fast-evolving sequences; and they display an ongoing fixation preference for G and C alleles. They are also significantly enriched for disease-associated polymorphisms, suggesting that they contribute to the fixation of deleterious alleles. The gBGC tracts provide a unique window into historical recombination processes along the human and chimpanzee lineages. They supply additional evidence of long-term conservation of megabase-scale recombination rates accompanied by rapid turnover of hotspots. Together, these findings shed new light on the evolutionary, functional, and disease implications of gBGC. The phastBias program and our predicted tracts are freely available. © 2013 Capra et al

The Twisted Magnetic Field of the Protobinary L483

We present H-band (1.65 μm) and SOFIA HAWC+ 154 μm polarization observations of the low-mass core L483. Our H-band observations reveal a magnetic field that is overwhelmingly in the E–W direction, which is approximately parallel to the bipolar outflow that is observed in scattered IR light and in single-dish 12CO observations. From our 154 μm data, we infer a ∼45° twist in the magnetic field within the inner 5″ (1000 au) of L483. We compare these new observations with published single-dish 350 μm polarimetry and find that the 10,000 au scale H-band data match the smaller-scale 350 μm data, indicating that the collapse of L483 is magnetically regulated on these larger scales. We also present high-resolution 1.3 mm Atacama Large Millimeter/submillimeter Array data of L483 that reveals it is a close binary star with a separation of 34 au. The plane of the binary of L483 is observed to be approximately parallel to the twisted field in the inner 1000 au. Comparing this result to the ∼1000 au protostellar envelope, we find that the envelope is roughly perpendicular to the 1000 au HAWC+ field. Using the data presented, we speculate that L483 initially formed as a wide binary and the companion star migrated to its current position, causing an extreme shift in angular momentum thereby producing the twisted magnetic field morphology observed. More observations are needed to further test this scenario

Re-Assembly of the Genome of Francisella tularensis Subsp. holarctica OSU18

Francisella tularensis is a highly infectious human intracellular pathogen that is the causative agent of tularemia. It occurs in several major subtypes, including the live vaccine strain holarctica (type B). F. tularensis is classified as category A biodefense agent in part because a relatively small number of organisms can cause severe illness. Three complete genomes of subspecies holarctica have been sequenced and deposited in public archives, of which OSU18 was the first and the only strain for which a scientific publication has appeared [1]. We re-assembled the OSU18 strain using both de novo and comparative assembly techniques, and found that the published sequence has two large inversion mis-assemblies. We generated a corrected assembly of the entire genome along with detailed information on the placement of individual reads within the assembly. This assembly will provide a more accurate basis for future comparative studies of this pathogen

Decreased Serologic Response in Vaccinated Military Recruits during 2011 Correspond to Genetic Drift in Concurrent Circulating Pandemic A/H1N1 Viruses

Population-based febrile respiratory illness surveillance conducted by the Department of Defense contributes to an estimate of vaccine effectiveness. Between January and March 2011, 64 cases of 2009 A/H1N1 (pH1N1), including one fatality, were confirmed in immunized recruits at Fort Jackson, South Carolina, suggesting insufficient efficacy for the pH1N1 component of the live attenuated influenza vaccine (LAIV).To test serologic protection, serum samples were collected at least 30 days post-vaccination from recruits at Fort Jackson (LAIV), Parris Island (LAIV and trivalent inactivated vaccine [TIV]) at Cape May, New Jersey (TIV) and responses measured against pre-vaccination sera. A subset of 78 LAIV and 64 TIV sera pairs from recruits who reported neither influenza vaccination in the prior year nor fever during training were tested by microneutralization (MN) and hemagglutination inhibition (HI) assays. MN results demonstrated that seroconversion in paired sera was greater in those who received TIV versus LAIV (74% and 37%). Additionally, the fold change associated with TIV vaccination was significantly different between circulating (2011) versus the vaccine strain (2009) of pH1N1 viruses (ANOVA p value = 0.0006). HI analyses revealed similar trends. Surface plasmon resonance (SPR) analysis revealed that the quantity, IgG/IgM ratios, and affinity of anti-HA antibodies were significantly greater in TIV vaccinees. Finally, sequence analysis of the HA1 gene in concurrent circulating 2011 pH1N1 isolates from Fort Jackson exhibited modest amino acid divergence from the vaccine strain.Among military recruits in 2011, serum antibody response differed by vaccine type (LAIV vs. TIV) and pH1N1 virus year (2009 vs. 2011). We hypothesize that antigen drift in circulating pH1N1 viruses contributed to reduce vaccine effectiveness at Fort Jackson. Our findings have wider implications regarding vaccine protection from circulating pH1N1 viruses in 2011-2012

Digital Signal Processing

Contains an introduction and reports on fourteen research projects.National Science Foundation FellowshipNational Science Foundation (Grant ECS84-07285)U.S. Navy - Office of Naval Research (Contract N00014-81-K-0742)Sanders Associates, Inc.U.S. Air Force - Office of Scientific Research (Contract F19628-85-K-0028)Advanced Television Research ProgramAmoco Foundation FellowshipHertz Foundation Fellowshi

Digital Signal Processing

Contains an introduction and reports on fifteen research projects.U.S. Navy - Office of Naval Research (Contract N00O14-81-K-0742)U.S. Navy - Office of Naval Research (Contract N00014-77-C-0266)National Science Foundation (Grant ECS80-07102)National Science Foundation (Grant ECS84-07285)Amoco Foundation FellowshipSanders Associates, Inc.Advanced Television Research ProgramM.I.T. Vinton Hayes FellowshipHertz Foundation Fellowshi

Digital Signal Processing

Contains introduction and reports on seventeen research projects.U.S. Navy - Office of Naval Research (Contract N00014-81-K-0742)U.S. Navy - Office of Naval Research (Contract N00014-77-C-0266)National Science Foundation (Grant ECS80-07102)Bell Laboratories FellowshipAmoco Foundation FellowshipSchlumberger-Doll Research Center FellowshipSanders Associates, Inc.Toshiba Company FellowshipM.I.T. Vinton Hayes FellowshipHertz Foundation Fellowshi

Digital Signal Processing Group

Contains an introduction and reports on nineteen research projects.U.S. Navy - Office of Naval Research (Contract N00014-77-C-0266)U.S. Navy - Office of Naval Research (Contract N00014-81-K-0742)National Science Foundation (Grant ECS80-07102)Bell Laboratories FellowshipAmoco Foundation FellowshipU.S. Navy - Office of Naval Research (Contract N00014-77-C-0196)Schlumberger-Doll Research Center FellowshipToshiba Company FellowshipVinton Hayes FellowshipHertz Foundation Fellowshi