RNA folding kinetics regulates translation of phage MS2 maturation gene

The gene for the maturation protein of the single-stranded RNA coliphage MS2 is preceded by an untranslated leader of 130 nt, which folds into a cloverleaf, i.e., three stem–loop structures enclosed by a long distance interaction (LDI). This LDI prevents translation because its 3′ moiety contains the Shine–Dalgarno sequence of the maturation gene. Previously, several observations suggested that folding of the cloverleaf is kinetically delayed, providing a time window for ribosomes to access the RNA. Here we present direct evidence for this model. In vitro experiments show that ribosome binding to the maturation gene is faster than refolding of the denatured cloverleaf. This folding delay appears related to special properties of the leader sequence. We have replaced the three stem–loop structures by a single five nt loop. This change does not affect the equilibrium structure of the LDI. Nevertheless, in this construct, the folding delay has virtually disappeared, suggesting that now the RNA folds faster than ribosomes can bind. Perturbation of the cloverleaf by an insertion makes the maturation start permanently accessible. A pseudorevertant that evolved from an infectious clone carrying the insertion had overcome this defect. It showed a wild-type folding delay before closing down the maturation gene. This experiment reveals the biological significance of retarded cloverleaf formation.

Chloroplast DNA footprints of postglacial recolonization by oaks

Recolonization of Europe by forest tree species after the last glaciation is well documented in the fossil pollen record. This spread may have been achieved at low densities by rare events of long-distance dispersal, rather than by a compact wave of advance, generating a patchy genetic structure through founder effects. In long-lived oak species, this structure could still be discernible by using maternally transmitted genetic markers. To test this hypothesis, a fine-scale study of chloroplast DNA (cpDNA) variability of two sympatric oak species was carried out in western France. The distributions of six cpDNA length variants were analyzed at 188 localities over a 200 × 300 km area. A cpDNA map was obtained by applying geostatistics methods to the complete data set. Patches of several hundred square kilometers exist which are virtually fixed for a single haplotype for both oak species. This local systematic interspecific sharing of the maternal genome strongly suggests that long-distance seed dispersal events followed by interspecific exchanges were involved at the time of colonization, about 10,000 years ago.

Differential Distribution of Dynamin Isoforms in Mammalian Cells

Dynamins are 100-kDa GTPases that are essential for clathrin-coated vesicle formation during receptor-mediated endocytosis. To date, three different dynamin genes have been identified, with each gene expressing at least four different alternatively spliced forms. Currently, it is unclear whether these different dynamin gene products perform distinct or redundant cellular functions. Therefore, the focus of this study was to identify additional spliced variants of dynamin from rat tissues and to define the distribution of the dynamin family members in a cultured rat epithelial cell model (Clone 9 cells). After long-distance reverse transcription (RT)-PCR of mRNA from different rat tissues, the full-length cDNAs encoding the different dynamin isoforms were sequenced and revealed four additional spliced variants for dynamin I and nine for dynamin III. Thus, in rat tissues there are a total of at least 25 different mRNAs produced from the three dynamin genes. Subsequently, we generated stably transfected Clone 9 cells expressing full-length cDNAs of six different spliced forms tagged with green fluorescent protein. Confocal or fluorescence microscopy of these transfected cells revealed that many of the dynamin proteins associate with distinct membrane compartments, which include clathrin-coated pits at the plasma membrane and the Golgi apparatus, and several undefined vesicle populations. These results indicate that the dynamin family is more extensive than was originally predicted and suggest that the different dynamin proteins are localized to distinct cytoplasmic or membrane compartments.

Explaining the moon illusion

An old explanation of the moon illusion holds that various cues place the horizon moon at an effectively greater distance than the elevated moon. Although both moons have the same angular size, the horizon moon must be perceived as larger. More recent explanations hold that differences in accommodation or other factors cause the elevated moon to appear smaller. As a result of this illusory difference in size, the elevated moon appears to be more distant than the horizon moon. These two explanations, both based on the geometry of stereopsis, lead to two diametrically opposed hypotheses. That is, a depth interval at a long distance is associated with a smaller binocular disparity, whereas an equal depth interval at a smaller distance is associated with a larger disparity. We conducted experiments involving artificial moons and confirmed the hypothesis that the horizon moon is at a greater perceptual distance. Moreover, when a moon of constant angular size was moved closer it was also perceived as growing smaller, which is consistent with the older explanation. Although Emmert's law does not predict the size–distance relationship over long distances, we conclude that the horizon moon is perceived as larger because the perceptual system treats it as though it is much farther away. Finally, we observe that recent explanations substitute perceived size for angular size as a cue to distance. Thus, they imply that perceptions cause perceptions.

Molecular evidence for an African origin of the Hawaiian endemic Hesperomannia (Asteraceae)

Identification of the progenitors of plants endemic to oceanic islands often is complicated by extreme morphological divergence between island and continental taxa. This is especially true for the Hawaiian Islands, which are 3,900 km from any continental source. We examine the origin of Hesperomannia, a genus of three species endemic to Hawaii that always have been placed in the tribe Mutisieae of the sunflower family. Phylogenetic analyses of representatives from all tribes in this family using the chloroplast gene ndhF (where ndhF is the ND5 protein of chloroplast NADH dehydrogenase) indicate that Hesperomannia belongs to the tribe Vernonieae. Phylogenetic comparisons within the Vernonieae using sequences of both ndhF and the internal transcribed spacer regions of nuclear ribosomal DNA reveal that Hesperomannia is sister to African species of Vernonia. Long-distance dispersal northeastward from Africa to southeast Asia and across the many Pacific Ocean island chains is the most likely explanation for this unusual biogeographic connection. The 17- to 26-million-year divergence time between African Vernonia and Hesperomannia estimated by the DNA sequences predates the age of the eight existing Hawaiian Islands. These estimates are consistent with an hypothesis that the progenitor of Hesperomannia arrived at one of the low islands of the Hawaiian-Emperor chain between the late Oligocene and mid-Miocene when these islands were above sea level. Subsequent to its arrival the southeast Pacific island chains served as steppingstones for dispersal to the existing Hawaiian Islands.

Phloem sap proteins from Cucurbita maxima and Ricinus communis have the capacity to traffic cell to cell through plasmodesmata

In angiosperms, the functional enucleate sieve tube system of the phloem appears to be maintained by the surrounding companion cells. In this study, we tested the hypothesis that polypeptides present within the phloem sap traffic cell to cell from the companion cells, where they are synthesized, into the sieve tube via plasmodesmata. Coinjection of fluorescently labeled dextrans along with size-fractionated Cucurbita maxima phloem proteins, ranging in size from 10 to 200 kDa, as well as injection of individual fluorescently labeled phloem proteins, provided unambiguous evidence that these proteins have the capacity to interact with mesophyll plasmodesmata in cucurbit cotyledons to induce an increase in size exclusion limit and traffic cell to cell. Plasmodesmal size exclusion limit increased to greater than 20 kDa, but less than 40 kDa, irrespective of the size of the injected protein, indicating that partial protein unfolding may be a requirement for transport. A threshold concentration in the 20–100 nM range was required for cell-to-cell transport indicating that phloem proteins have a high affinity for the mesophyll plasmodesmal binding site(s). Parallel experiments with glutaredoxin and cystatin, phloem sap proteins from Ricinus communis, established that these proteins can also traffic through cucurbit mesophyll plasmodesmata. These results are discussed in terms of the requirements for regulated protein trafficking between companion cells and the sieve tube system. As the threshold value for plasmodesmal transport of phloem sap proteins falls within the same range as many plant hormones, the possibility is discussed that some of these proteins may act as long-distance signaling molecules.

Biogeographic range expansion into South America by Coccidioides immitis mirrors New World patterns of human migration

Long-distance population dispersal leaves its characteristic signature in genomes, namely, reduced diversity and increased linkage between genetic markers. This signature enables historical patterns of range expansion to be traced. Herein, we use microsatellite loci from the human pathogen Coccidioides immitis to show that genetic diversity in this fungus is geographically partitioned throughout North America. In contrast, analyses of South American C. immitis show that this population is genetically depauperate and was founded from a single North American population centered in Texas. Variances of allele distributions show that South American C. immitis have undergone rapid population growth, consistent with an epidemic increase in postcolonization population size. Herein, we estimate the introduction into South America to have occurred within the last 9,000–140,000 years. This range increase parallels that of Homo sapiens. Because of known associations between Amerindians and this fungus, we suggest that the colonization of South America by C. immitis represents a relatively recent and rapid codispersal of a host and its pathogen.

Nicotianamine Chelates Both FeIII and FeII. Implications for Metal Transport in Plants1

Nicotianamine (NA) occurs in all plants and chelates metal cations, including FeII, but reportedly not FeIII. However, a comparison of the FeII and ZnII affinity constants of NA and various FeIII-chelating aminocarboxylates suggested that NA should chelate FeIII. High-voltage electrophoresis of the FeNA complex formed in the presence of FeIII showed that the complex had a net charge of 0, consistent with the hexadentate chelation of FeIII. Measurement of the affinity constant for FeIII yielded a value of 1020.6, which is greater than that for the association of NA with FeII (1012.8). However, capillary electrophoresis showed that in the presence of FeII and FeIII, NA preferentially chelates FeII, indicating that the FeIINA complex is kinetically stable under aerobic conditions. Furthermore, Fe complexes of NA are relatively poor Fenton reagents, as measured by their ability to mediate H2O2-dependent oxidation of deoxyribose. This suggests that NA will have an important role in scavenging Fe and protecting the cell from oxidative damage. The pH dependence of metal ion chelation by NA and a typical phytosiderophore, 2′-deoxymugineic acid, indicated that although both have the ability to chelate Fe, when both are present, 2′-deoxymugineic acid dominates the chelation process at acidic pH values, whereas NA dominates at alkaline pH values. The consequences for the role of NA in the long-distance transport of metals in the xylem and phloem are discussed.

Chloroplast-Avoidance Response Induced by High-Fluence Blue Light in Prothallial Cells of the Fern Adiantum capillus-veneris as Analyzed by Microbeam Irradiation1

Chloroplast movement was induced by partial cell illumination using a high-fluence blue microbeam in light-grown and dark-adapted prothallial cells of the fern Adiantum capillus-veneris. Chloroplasts inside the illuminated area moved out (high-fluence response [HFR]), whereas those outside moved toward the irradiated area (low-fluence response [LFR]), although they stopped moving when they reached the border. These results indicate that both HFR and LFR signals are generated by high-fluence blue light of the same area, and that an LFR signal can be transferred long-distance from the beam spot, although an HFR signal cannot. The lifetime of the HFR signal was calculated from the traces of chloroplast movement induced by a brief pulse from a high-fluence blue microbeam to be about 6 min. This is very short compared with that of the LFR (30–40 min; T. Kagawa, M. Wada [1994] J Plant Res 107: 389–398). These data indicate that the signal transduction pathways of the HFR and the LFR must be distinct.

Systemic spread of an RNA insect virus in plants expressing plant viral movement protein genes

Flock house virus (FHV), a single-stranded RNA insect virus, has previously been reported to cross the kingdom barrier and replicate in barley protoplasts and in inoculated leaves of several plant species [Selling, B. H., Allison, R. F. & Kaesberg, P. (1990) Proc. Natl. Acad. Sci. USA 87, 434–438]. There was no systemic movement of FHV in plants. We tested the ability of movement proteins (MPs) of plant viruses to provide movement functions and cause systemic spread of FHV in plants. We compared the growth of FHV in leaves of nontransgenic and transgenic plants expressing the MP of tobacco mosaic virus or red clover necrotic mosaic virus (RCNMV). Both MPs mobilized cell-to-cell and systemic movement of FHV in Nicotiana benthamiana plants. The yield of FHV was more than 100-fold higher in the inoculated leaves of transgenic plants than in the inoculated leaves of nontransgenic plants. In addition, FHV accumulated in the noninoculated upper leaves of both MP-transgenic plants. RCNMV MP was more efficient in mobilizing FHV to noninoculated upper leaves. We also report here that FHV replicates in inoculated leaves of six additional plant species: alfalfa, Arabidopsis, Brassica, cucumber, maize, and rice. Our results demonstrate that plant viral MPs cause cell-to-cell and long-distance movement of an animal virus in plants and offer approaches to the study of the evolution of viruses and mechanisms governing mRNA trafficking in plants as well as to the development of promising vectors for transient expression of foreign genes in plants.

Ediacaran biota from Sonora, Mexico.

The Ediacaran biota is the earliest diverse community of macroscopic animals and protoctists. Body and trace fossils in the Clemente Formation of northwestern Sonora extend downward the geologic range of Ediacaran forms. Taxa present in the Clemente Formation include cf. Cyclomedusa plana, Sekwia sp., an erniettid (bearing an air mattress-like "pneu" body construction), and the trace fossils Lockeia ichnosp. and Palaeophycus tubularis. The trace fossils confirm the presence of sediment-dwelling animals in this shallow marine community. The body fossils are headless, tailless, and appendageless. Some may be body fossils of animals but others may be fossils of large protoctists. These body and trace fossils, recovered from thinly bedded sandstones and siltstones, occur 75 meters lower in the Sonoran stratigraphic section than a distinctive Clemente Formation oolite. The stratigraphic position of the fossils below this oolite permits long-distance correlation between fossiliferous Proterozoic strata of Mexico and the United States. Correlations utilizing both the Clemente Formation oolite and a trace fossil (Vermiforma antiqua) confirm the antiquity (600 million years or more) of this body fossil-rich community of macroscopic eukaryotes. The recently discovered body fossils are the oldest known remains of the Ediacaran biota.

Uso de redes sociales para mejorar el rendimiento de los alumnos con diferentes estilos de aprendizaje

Debido a los cambios que el Espacio Europeo de Educación Superior introduce al potenciar las horas de trabajo no presencial, se hacen necesarios nuevos mecanismos para posibilitar una mejor comunicación y cooperación en el proceso de aprendizaje. Las redes sociales, como Facebook, pueden suministrar estos mecanismos, pero su uso satisfactorio para la docencia puede verse afectado en gran medida por el estilo de aprendizaje de los alumnos. Este artículo plantea la necesidad de estudiar la influencia de los diferentes estilos de aprendizaje en la docencia no presencial mediante el uso de redes sociales con el fin de incrementar el rendimiento de los alumnos. Cabe destacar que este artículo describe el proyecto “Las redes sociales y su relación con los estilos de aprendizaje” a realizar dentro del programa de Redes de Investigación en Docencia Universitaria del Instituto de Ciencias de la Educación de la Universidad de Alicante.

Performing Pedagogies and Exploring Embodied Knowledge

Performing Pedagogies was a week-long performance and exhibition series I organized that took place in Kingston, Ontario between March 15th - March 20th 2016. The motivation for this project came from a desire to explore performative modes of experiencing critical, embodied knowledge. The series featured five performances, a long distance collaboration between thirty-one Queen’s undergraduate students and a Vancouver artist-run free school (The School for Eventual Vacancy), a subsequent exhibition, a panel discussion, and a radical performance pedagogy workshop led by co-artistic director of the international performance art troupe, La Pocha Nostra. Artists featured included Golboo Amani, Basil AlZeri, Caitlin Chaisson, Justin Langlois, Saul Garcia-Lopez, Francisco-Fernando Granados, and Andrew Rabyniuk. By curating examples of performance art that variously incorporated embodied pedagogical interventions, I examined the processes of performance as pedagogy. Performing Pedagogies explored interventions into contemporary contours of neoliberal education paradigms through embodied encounters—fostering conversations about the meanings and limitations of knowledge dissemination and education today and posing questions about possibilities for radical pedagogies, embodied knowledge, and counter curricula.

Performing Pedagogies and Exploring Embodied Knowledge as Curatorial Investigation

Performing Pedagogies was a week-long performance and exhibition series I organized that took place in Kingston, Ontario between March 15th - March 20th 2016. The motivation for this project came from a desire to explore performative modes of experiencing critical, embodied knowledge. The series featured five performances, a long distance collaboration between thirty-one Queen’s undergraduate students and a Vancouver artist-run free school (The School for Eventual Vacancy), a subsequent exhibition, a panel discussion, and a radical performance pedagogy workshop led by co-artistic director of the international performance art troupe, La Pocha Nostra. Artists featured included Golboo Amani, Basil AlZeri, Caitlin Chaisson, Justin Langlois, Saul Garcia-Lopez, Francisco-Fernando Granados, and Andrew Rabyniuk. By curating examples of performance art that variously incorporated embodied pedagogical interventions, I examined the processes of performance as pedagogy. Performing Pedagogies explored interventions into contemporary contours of neoliberal education paradigms through embodied encounters—fostering conversations about the meanings and limitations of knowledge dissemination and education today and posing questions about possibilities for radical pedagogies, embodied knowledge, and counter curricula.

Developmental control of H+/amino acid permease gene expression during seed development of Arabidopsis

Long distance transport of amino acids is mediated by several families of differentially expressed amino acid transporters. The two genes AAP1 and AAP2 encode broad specificity H+-amino acid co-transporters and are expressed to high levels in siliques of Arabidopsis, indicating a potential role in supplying the seeds with organic nitrogen. The expression of both genes is developmentally controlled and is strongly induced in siliques at heart stage of embryogenesis, shortly before induction of storage protein genes. Histochemical analysis of transgenic plants expressing promoter-GUS fusions shows that the genes have non-overlapping expression patterns in siliques. AAP1 is expressed in the endosperm and the cotyledons whereas AAP2 is expressed in the vascular strands of siliques and in funiculi. The endosperm expression of AAP1 during early stages of seed development indicates that the endosperm serves as a transient storage tissue for organic nitrogen. Amino acids are transported in both xylem and phloem but during seed filling are imported only via the phloem. AAP2, which is expressed in the phloem of stems and in the veins supplying seeds, may function in uptake of amino acids assimilated in the green silique tissue, in the retrieval of amino acids leaking passively out of the phloem and in xylem-to-phloem transfer along the path. The promoters provide excellent tools to study developmental, hormonal and metabolic control of nitrogen nutrition during development and may help to manipulate the timing and composition of amino acid import into seeds.