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Made available in DSpace on 2021-06-25T10:57:46Z (GMT). No. of bitstreams: 0 Previous issue date: 2021-06-01 Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) The mechanism of extracellular traps (ETs) is important in the cellular response against bacteria. Thus, in the present study, we describe for the first time the capacity of the Nile tilapia (Oreochromis niloticus) microglia in the formation of ETs in Weissella cibaria in vitro infection. Thus, we evaluated the ultrastructure of the microglia culture and observed the formation of ETs 6 h after stimulation with lipopolysaccharide (LPS) and during the course of infection. Our results shed light on the mechanism of formation of ETs in the microglia of teleost fish and the ability of W. cibaria to infect these cells. Department of Postgraduate in Health Sciences-PROCISA Federal University of Roraima (UFRR) Immunochemistry Laboratory Butantan Institute (CeTICS/FAPESP) Department of Technology School of Agrarian and Veterinary Sciences Sao Paulo State University (Unesp) Laboratory of Animal Pharmacology and Toxicology University of Brazil, Descalvado Department of Preventive Veterinary Medicine of Unesp School of Agrarian and Veterinary Sciences São Paulo State University (Unesp), Jaboticabal Department of Technology School of Agrarian and Veterinary Sciences Sao Paulo State University (Unesp) Department of Preventive Veterinary Medicine of Unesp School of Agrarian and Veterinary Sciences São Paulo State University (Unesp), Jaboticabal

PEA3 transcription factor subfamily is present in a variety of tissues with branching morphogenesis, and play a particularly significant role in neural circuit formation and specificity. Many target genes in axon guidance and cell–cell adhesion pathways have been identified for Pea3 transcription factor (but not for Erm or Er81); however it was not so far clear whether all Pea3 subfamily members regulate same target genes, or whether there are unique targets for each subfamily member that help explain the exclusivity and specificity of these proteins in neuronal circuit formation. In this study, using transcriptomics and qPCR analyses in SH-SY5Y neuroblastoma cells, hypothalamic and hippocampal cell line, we have identified cell type-specific and subfamily member-specific targets for PEA3 transcription factor subfamily. While Pea3 upregulates transcription of Sema3D and represses Sema5B, for example, Erm and Er81 upregulate Sema5A and Er81 regulates Unc5C and Sema4G while repressing EFNB3 in SH-SY5Y neuroblastoma cells. We furthermore present a molecular model of how unique sites within the ETS domain of each family member can help recognize specific target motifs. Such cell-context and member-specific combinatorial expression profiles help identify cell–cell and cell-extracellular matrix communication networks and how they establish specific connections.

The genus Artemisia L. of the family Asteraceae is systematically very complex. The aim of this study was to evaluate taxonomic positions of taxa of the subgenus Artemisia belonging to the genus Artemisia in Turkey using some molecular techniques. In this molecular study, 44 individuals belong to 14 species of the subgenus Artemisia were examined. Analyses were performed on the combined dataset using maximum parsimony, maximum likelihood and Bayesian inference and Molecular parameters obtained from co-evaluations of sequences of the psbA-trnH, ITS and ETS regions of examined individuals were used in the phylogenetic tree drawing. According to the results of this study, two molecular groups have been formed based on the DNA sequence similarity of the species, but there are no obvious morphological characters corresponding to two molecular groups. There is no also agreement between the two molecular groups and the two morphological groups formed according to the hairiness condition of the receptacle of species. Due to the lack of molecular significance of their receptacles with or without hair, dividing of the subgenus Artemisia species into new subgenera or sections was not considered appropriate. Likewise, it has been found that with or without hair on the corolla lobes of the central hermaphrodite disc flowers have no molecular significance. It was found that there were no gene flow and hybridization between the 14 species of the subgenus Artemisia and these 14 species were found completed their speciation. This study is important as it is the first molecular based study relating with belong to subgenus Artemisia species growing naturally in Turkey. In addition, new haplotypes related to the populations of Turkey belonging to the subgenus Artemisia taxa were reported by us for the first time and added to the GenBank database. Resumo O gênero Artemisia L. da família Asteraceae é sistematicamente muito complexo. O objetivo deste estudo foi avaliar as posições taxonômicas de táxons do subgênero Artemisia pertencentes ao gênero Artemisia na Turquia usando algumas técnicas moleculares. Neste estudo molecular, 44 indivíduos pertencentes a 14 espécies do subgênero Artemisia foram examinados. As análises foram realizadas no conjunto de dados combinado usando máxima parcimônia, máxima verossimilhança e inferência bayesiana e parâmetros moleculares obtidos a partir de coavaliações de sequências das regiões psbA-trnH, ITS e ETS de indivíduos examinados foram usados no desenho da árvore filogenética. De acordo com os resultados deste estudo, dois grupos moleculares foram formados com base na similaridade da sequência de DNA das espécies, mas não há caracteres morfológicos óbvios correspondentes a dois grupos moleculares. Também não há concordância entre os dois grupos moleculares e os dois grupos morfológicos formados de acordo com a condição de pilosidade do receptáculo da espécie. Devido à falta de significado molecular de seus receptáculos com ou sem cabelo, a divisão das espécies do subgênero Artemisia em novos subgêneros ou seções não foi considerada apropriada. Da mesma forma, verificou-se que com ou sem cabelo nos lobos da corola das flores do disco hermafrodita central não tem significado molecular. Constatou-se que não houve fluxo gênico e hibridização entre as 14 espécies do subgênero Artemisia e essas 14 espécies concluíram sua especiação. Este estudo é importante porque é o primeiro estudo de base molecular relacionado com espécies pertencentes ao subgênero Artemisia crescendo naturalmente na Turquia. Além disso, novos haplótipos relacionados às populações da Turquia pertencentes ao subgênero Artemisia taxa foram relatados por nós pela primeira vez e adicionados ao banco de dados do GenBank.

ETS family transcription factors play major roles in prostate tumorigenesis with some acting as oncogenes and others as tumor suppressors. ETS factors can compete for binding at some cis-regulatory sequences, but display specific binding at others. Therefore, changes in expression of ETS family members during tumorigenesis can have complex, multimodal effects. Here we show that ELF1 was the most commonly down-regulated ETS factor in primary prostate tumors, and expression decreased further in metastatic disease. Genome-wide mapping in cell lines indicated that ELF1 has two distinct tumor suppressive roles mediated by distinct cis-regulatory sequences. First, ELF1 inhibited cell migration and epithelial-mesenchymal transition by interfering with oncogenic ETS functions at ETS/AP-1 cis-regulatory motifs. Second, ELF1 uniquely targeted and activated genes that promote senescence. Furthermore, knockdown of ELF1 increased docetaxel resistance, indicating that the genomic deletions found in metastatic prostate tumors may promote therapeutic resistance through loss of both RB1 and ELF1.

Mutant p53(s) are widely considered as oncogenes and promote several gain-of-function oncogenic activities. p53 mutations correlate with higher rates of metastasis and poor survival, therefore, it is paramount to inhibit mutant p53 protein either directly or indirectly. Although some compounds have been developed, none of them have achieved a desirable level of specificity. Some of these compounds only targeted specific mutations. In search of less-toxic compounds, we tested plant-derived compounds on mutant p53 triple-negative breast cancer cell lines. Here, we show that the compounds tested reduced the protein levels of one of the more frequent oncogenic p53 mutants (R249S, hot spot mutation), and its important targets that promote invasion and metastasis, including GMPS and IMPDH1. All compounds tested perturbed the invasion potential of the breast cancer cell line. These compounds downregulated several nucleotide metabolism genes (NMGs) which are essential for cell cycle progression. We observed S-phase arrest correlating to reduced cell proliferation and increased replication stress. Moreover, we also show a reduction of key ETS transcription family members including ETS2, ETS1, ETV1, and ETV4, which are involved in invasion and metastasis. We propose that these compounds may inhibit invasion by interfering with multiple pathways. Our findings exemplify that these tested compounds could inhibit invasion and cell growth in TNBC in a nucleotide-dependent manner.

The ETS domain family of transcription factors is involved in a number of biological processes, and is commonly misregulated in various forms of cancer. Using microarray datasets from patients with different grades of glioma, we have analyzed the expression profiles of various ETS genes, and have identified ETV1, ELK3, ETV4, ELF4, and ETV6 as novel biomarkers for the identification of different glioma grades. We have further analyzed the gene regulatory networks of ETS transcription factors and compared them to previous microarray studies, where Elk-1-VP16 or PEA3-VP16 were overexpressed in neuroblastoma cell lines, and we identify unique and common regulatory networks for these ETS proteins.

Correct vascular differentiation requires distinct patterns of gene expression in different subtypes of endothelial cells. Members of the ETS transcription factor family are essential for the transcriptional activation of arterial and angiogenesis-specific gene regulatory elements, leading to the hypothesis that they play lineage-defining roles in arterial and angiogenic differentiation directly downstream of VEGFA signalling. However, an alternative explanation is that ETS binding at enhancers and promoters is a general requirement for activation of many endothelial genes regardless of expression pattern, with subtype-specificity provided by additional factors. Here we use analysis of Ephb4 and Coup-TFII (Nr2f2) vein-specific enhancers to demonstrate that ETS factors are equally essential for vein, arterial and angiogenic-specific enhancer activity patterns. Further, we show that ETS factor binding at these vein-specific enhancers is enriched by VEGFA signalling, similar to that seen at arterial and angiogenic enhancers. However, while arterial and angiogenic enhancers can be activated by VEGFA in vivo, the Ephb4 and Coup-TFII venous enhancers are not, suggesting that the specificity of VEGFA-induced arterial and angiogenic enhancer activity occurs via non-ETS transcription factors. These results support a model in which ETS factors are not the primary regulators of specific patterns of gene expression in different endothelial subtypes., Highlights • Vein-specific enhancers can contain essential ETS motifs. • VEGFA induced an increase in ETS binding at vein, arterial and angiogenic enhancers. • VEGFA stimulation cannot induce vein-specific enhancer activity.

Tephroseris is generally considered a difficult genus. Based on the examination of extensive herbarium material and considering the existing literature, we recognize seven species in Europe outside Russia. These are T. palustris, T. integrifolia with subsp. integrifolia, subsp. aurantiaca, subsp. capitata, subsp. maritima, subsp. serpentini and subsp. “tundricola”, T. balbisiana, T. crispa, T. helenitis, T. longifolia and T. papposa. Phylogenetic analysis of ITS and ETS sequences showed that these species fall into three lineages. These are: (1) T. palustris, clearly related to Arctic species of the genus; (2) T. integrifolia; and (3) the remaining species. Molecular dating of the T. integrifolia lineage resulted in a crown group age of 1.76 (0.85–2.87) million years. Possible reasons for taxonomic difficulties in the genus in Europe outside Russia may include its young phylogenetic age and extensive migration and genetic admixture in the Quaternary. The decline of the genus in Europe outside Russia is documented and discussed. We consider it possible that its decline is related to rising global temperatures. Citation: Kadereit J. W., Laux P. & Dillenberger M. S. 2021: A conspectus of Tephroseris (Asteraceae: Senecioneae) in Europe outside Russia and notes on the decline of the genus. – Willdenowia 51: 271–317. Version of record first published online on 3 September 2021 ahead of inclusion in August 2021 issue.

The signal regulated transcription factors (SRTFs) control the ultimate transcriptional output of signaling pathways. Here, we examined a family of FGF-induced SRTFs – Etv1, Etv 4, and Etv 5 – in murine lens development. Contrary to FGF receptor mutants that displayed loss of ERK signaling and defective cell differentiation, Etv deficiency augmented ERK phosphorylation without disrupting the normal lens fiber gene expression. Instead, the transitional zone for lens differentiation was shifted anteriorly as a result of reduced Jag1-Notch signaling. We also showed that Etv proteins suppresses mTOR activity by promoting Tsc2 expression, which is necessary for the nuclei clearance in mature lens. These results revealed the functional divergence between Etv and FGF in lens development, demonstrating that these SRTFs can operate outside the confine of their upstream signaling., eLife digest Many cells contain proteins known as signal-induced transcription factors, which are poised to receive messages from the environment and then react by activating genes required for the cell to respond appropriately. It is commonly thought that these transcription factors faithfully follow the instructions they receive from the external signal: for instance, if the message was to encourage the cell to grow, the transcription factors would switch on growth-related genes. As the eyes of mice and other mammals develop, a signal known as FGF is required for certain cells to specialize into lens fiber cells: these long, thin, transparent cells form the bulk of the lens, the structure that allows focused vision. Previous studies suggest that FGF activates three transcription factors known as Etv1, Etv4 and Etv5, but their precise roles in the development of the lens has remained unclear. Here, Garg, Hannan, Wang et al. confirm that FGF signaling does indeed activate all three proteins. However, mutant mice that lacked Etv1, Etv4 and Etv5 still created lens fiber cells, suggesting that the transcription factors are largely unnecessary for lens fiber cells formation. Instead, the Etv proteins participated in a cascade of molecular events involving a protein called Notch; as a result, if the transcription factors were absent, the lens fiber cells formed prematurely. In addition, deactivating Etv1, Etv4 and Etv5 also promoted the activity of a protein which interfered with the removal of internal cell compartments, a process required for lens fiber cells to mature properly. These findings reveal that the roles of Etv1, Etv4 and Etv5 deviate from and even oppose FGF signaling in the lenses of mice. Transcription factors control the ultimate fate of a cell, and there is therefore increased interest in targeting them for therapy. The work by Garg, Hannan, Wang et al. reveals an unexpected complexity in how these proteins respond to upstream signals, highlighting the importance of further dissecting these relationships.

International audience; Specification of the main axes of polarity of the embryo is an essential process during embryonic development. In many species, this process is achieved by the localization of maternal factors into discrete regions of the egg. However, in other animals, like in amniotes and in echinoderms, the considerable plasticity of the early blastomeres seems to preclude the existence of maternal determinants and the mechanisms by which the radial symmetry of the egg is broken remain largely enigmatic. In this chapter, we review recent progress on the identification of maternal components involved in symmetry breaking and dorsal-ventral (D/V) axis formation of the sea urchin embryo. We will first review some key experiments on D/V axis formation from classical embryologists that provided evidence for a weak maternal D/V prepattern. We will then detail more recent molecular analyses that established the critical role played by Nodal signaling in allocating cell fates along the secondary axis and led to the discovery that maternal transcription factors such as the Sry-related HMG box B1 (SoxB1), the Octamer binding factor1/2 (Oct1/2), the T-cell factor/Lymphoid enhancer-binding factor (TCF/LEF) and the Erythroblastosis virus E26 Oncogene Homolog (ETS) domain transcriptional repressor Translocation-Ets-Leukemia virus protein (Yan/Tel) as well as maternal signaling molecules like Univin are essential for the initiation of nodal expression. Finally, we will describe recent advances that uncovered a role in symmetry breaking and dorsal-ventral axis orientation for the transforming growth factor beta (TGF-beta)-like factor Panda, which appears to be both necessary and sufficient for D/V axis orientation. Therefore, even in the highly regulative sea urchin embryo, the activity of localized maternal factors provides the embryo with a blueprint of the D/V axis.

Plankton community respiration (R) is a major component of the carbon flux in aquatic ecosystems. However, current methods to measure actual respiration from oxygen consumption at relevant spatial scales are not sensitive enough in oligotrophic environments where respiration rates are very low. To overcome this drawback, more sensitive indirect enzymatic approaches are commonly used as R proxies. The in vivo electron transport system (ETSvivo) assay, which measures the reduction of (2-(4-iodophenyl)-3-(4-nitrophenyl)-5-phenyl tetrazolium chloride salt, INT) to INT-formazan in the presence of natural substrate levels, was recently proposed as an indirect reliable estimation of R for natural plankton communities. However, under in vivo conditions, formazan salts could be toxic to the cells. Here, we test the toxicity of 0.2 mM of final INT concentration, widely used for ETSvivo assays, on natural bacterial assemblages collected in coastal and oceanic waters off Gran Canaria (Canary Islands, subtropical North Atlantic), in eight independent experiments. After 0.5 h of incubation, a significant but variable decline in cell viability (14–49%) was observed in all samples inoculated with INT. Moreover, INT also inhibited leucine uptake in less than 90 min of incubation. In the light of these results, we argue that enzymatic respiratory rates obtained with the ETSvivo method need to be interpreted with caution to derive R in oceanic regions where bacteria largely contribute to community respiration. Moreover, the variable toxicity on bacterial assemblages observed in our experiments questions the use of a single R/ETSvivo relationship as a universal proxy for regional studies.

Plant-pathogenic Xanthomonas infects a wide variety of host plants and causes many devastating diseases on crops. Transcription activator-like effectors (TALEs) are delivered by a type III secretion system (T3SS) of Xanthomonas into plant nuclei to directly bind specific DNA sequences (TAL effector-binding elements, EBEs) on either strand of host target genes with an unique modular DNA-binding domain and to bidirectionally drive host gene transcription. The target genes in plants consist of host susceptibility (S) genes promoting disease (ETS) and resistance (R) genes triggering defense (ETI). Here we generally summarized the discovery of TALEs in Xanthomonas species, their functions in bacterial pathogenicity in plants and their target genes in different host plants, and then focused on the newly revealed modes of protein action in triggering or suppressing plant defense.

Circumscriptions of and relationships among many genera and suprageneric taxa of the diverse grass tribe Poeae remain controversial. In an attempt to clarify these, we conducted phylogenetic analyses of >2400 new DNA sequences from two nuclear ribosomal regions (ITS, including internal transcribed spacers 1 and 2 and the 5.8S gene, and the 3'-end of the external transcribed spacer (ETS)) and five plastid regions (matK, trnL–trnF, atpF–atpH, psbK–psbI, psbA–rps19–trnH), and of more than 1000 new and previously published ITS sequences, focused particularly on Poeae chloroplast group 1 and including broad and increased species sampling compared to previous studies. Deep branches in the combined plastid and combined ITS+ETS trees are generally well resolved, the trees are congruent in most aspects, branch support across the trees is stronger than in trees based on only ITS and fewer plastid regions, and there is evidence of conflict between data partitions in some taxa. In plastid trees, a strongly supported clade corresponds to Poeae chloroplast group 1 and includes Agrostidinae p.p., Anthoxanthinae, Aveninae s.str., Brizinae, Koeleriinae (sometimes included in Aveninae s.l.), Phalaridinae and Torreyochloinae. In the ITS+ETS tree, a supported clade includes these same tribes as well as Sesleriinae and Scolochloinae. Aveninae s.str. and Sesleriinae are sister taxa and form a clade with Koeleriinae in the ITS+ETS tree whereas Aveninae s.str. and Koeleriinae form a clade and Sesleriinae is part of Poeae chloroplast group 2 in the plastid tree. All species of Trisetum are part of Koeleriinae, but the genus is polyphyletic. Koeleriinae is divided into two major subclades: one comprises Avellinia, Gaudinia, Koeleria, Rostraria, Trisetaria and Trisetum subg. Trisetum, and the other Calamagrostis/Deyeuxia p.p. (multiple species from Mexico to South America), Peyritschia, Leptophyllochloa, Sphenopholis, Trisetopsis and Trisetum subg. Deschampsioidea. Graphephorum, Trisetum cernuum, T. irazuense and T. macbridei fall in different clades of Koeleriinae in plastid vs. nuclear ribosomal trees, and are likely of hybrid origin. ITS and matK trees identify a third lineage of Koeleriinae corresponding to Trisetum subsect. Sibirica, and affinities of Lagurus ovatus with respect to Aveninae s.str. and Koeleriinae are incongruent in nuclear ribosomal and plastid trees, supporting recognition of Lagurus in its own subtribe. A large clade comprises taxa of Agrostidinae, Brizinae and Calothecinae, but neither Agrostidinae nor Calothecinae are monophyletic as currently circumscribed and affinities of Brizinae differ in plastid and nuclear ribosomal trees. Within this clade, one newly identified lineage comprises Calamagrostis coarctata, Dichelachne, Echinopogon (Agrostidinae p.p.) and Relchela (Calothecinae p.p.), and another comprises Chascolytrum (Calothecinae p.p.) and Deyeuxia effusa (Agrostidinae p.p.). Within Agrostidinae p.p., the type species of Deyeuxia and Calamagrostis s.str. are closely related, supporting classification of Deyeuxia as a synonym of Calamagrostis s.str. Furthermore, the two species of Ammophila are not sister taxa and are nested among different groups of Calamagrostis s.str., supporting their classification in Calamagrostis. Agrostis, Lachnagrostis and Polypogon form a clade and species of each are variously intermixed in plastid and nuclear ribosomal trees. Additionally, all but one species from South America classified in Deyeuxia sect. Stylagrostis resolve in Holcinae p.p. (Deschampsia). The current phylogenetic results support recognition of the latter species in Deschampsia, and we also demonstrate Scribneria is part of this clade. Moreover, Holcinae is not monophyletic in its current circumscription because Deschampsia does not form a clade with Holcus and Vahlodea, which are sister taxa. The results support recognition of Deschampsia in its own subtribe Aristaveninae. Substantial further changes to the classification of these grasses will be needed to produce generic circumscriptions consistent with phylogenetic evidence. The following 15 new combinations are made: Calamagrostis × calammophila, C. breviligulata, C. breviligulata subsp. champlainensis, C. × don-hensonii, Deschampsia aurea, D. bolanderi, D. chrysantha, D. chrysantha var. phalaroides, D. eminens, D. eminens var. fulva, D. eminens var. inclusa, D. hackelii, D. ovata, and D. ovata var. nivalis. D. podophora; the new name Deschampsia parodiana is proposed; the new subtribe Lagurinae is described; and a second-step lectotype is designated for the name Deyeuxia phalaroides.

Ewing Sarcoma is the second most common solid pediatric malignant neoplasm of bone and soft tissue. Driven by EWS/Ets, or rarely variant, oncogenic fusions, Ewing Sarcoma is a biologically and clinically aggressive disease with a high propensity for metastasis. However, the mechanisms underpinning Ewing Sarcoma metastasis are currently not well understood. In the present study, we identify and characterize a novel metastasis-promotional pathway in Ewing Sarcoma, involving the histone demethylase KDM3A, previously identified by our laboratory as a new cancer-promoting gene in this disease. Using global gene expression profiling, we show that KDM3A positively regulates genes and pathways implicated in cell migration and metastasis, and demonstrate, using functional assays, that KDM3A promotes migration in vitro and experimental, post-intravasation, metastasis in vivo. We further identify the melanoma cell adhesion molecule (MCAM) as a novel KDM3A target gene in Ewing Sarcoma, and an important effector of KDM3A pro-metastatic action. Specifically, we demonstrate that MCAM depletion, like KDM3A depletion, inhibits cell migration in vitro and experimental metastasis in vivo, and that MCAM partially rescues impaired migration due to KDM3A knock-down. Mechanistically, we show that KDM3A regulates MCAM expression both through a direct mechanism, involving modulation of H3K9 methylation at the MCAM promoter, and an indirect mechanism, via the Ets1 transcription factor. Finally, we identify an association between high MCAM levels in patient tumors and poor survival, in two different Ewing Sarcoma clinical cohorts. Taken together, our studies uncover a new metastasis-promoting pathway in Ewing Sarcoma, with therapeutically targetable components.

In Caenorhabditis elegans, the JNK MAP kinase (MAPK) pathway is important for axon regeneration. The JNK pathway is activated by a signaling cascade consisting of the growth factor SVH‐1 and its receptor tyrosine kinase SVH‐2. Expression of the svh‐2 gene is induced by axonal injury in a process involving the transcription factors ETS‐4 and CEBP‐1. Here, we find that svh‐14/mxl‐1, a gene encoding a Max‐like transcription factor, is required for activation of svh‐2 expression in response to axonal injury. We show that MXL‐1 binds to and inhibits the function of TDPT‐1, a C. elegans homolog of mammalian tyrosyl‐DNA phosphodiesterase 2 [TDP2; also called Ets1‐associated protein II (EAPII)]. Deletion of tdpt‐1 suppresses the mxl‐1 defect, but not the ets‐4 defect, in axon regeneration. TDPT‐1 induces SUMOylation of ETS‐4, which inhibits ETS‐4 transcriptional activity, and MXL‐1 counteracts this effect. Thus, TDPT‐1 interacts with two different transcription factors in axon regeneration.

Chryselium Urtubey & S. E. Freire, a new monotypic genus, is here described and illustrated to accomodate one species that was initially placed in Helichrysum, H. gnaphalioides, as Chryselium gnaphalioides. The new genus, Chryselium, is a monoecious herb with single stems and terminal capitulescences that are densely glomerulate from the Tropical Andes of Venezuela, Colombia, Ecuador, Bolivia, and Peru. This genus differs fundamentally from Helichrysum by having a stoloniferous habit and corollas with purple apices. Amolecular analysis using DNA sequences from nuclear (ITS and ETS) markers placed this genus in a clade with Chevreulia, Cuatrecasasiella, Antennaria, and Diaperia in the FLAG group. Affinities of the new genus with genera previously assigned to Chryselium are also discussed. Fil: Freire, Susana Edith. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Botánica Darwinion. Academia Nacional de Ciencias Exactas, Físicas y Naturales. Instituto de Botánica Darwinion; Argentina Fil: Urtubey, Estrella. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Botánica Darwinion. Academia Nacional de Ciencias Exactas, Físicas y Naturales. Instituto de Botánica Darwinion; Argentina

Background DNA methylation is involved in the regulation of gene expression. Although bisulfite-sequencing based methods profile DNA methylation at a single CpG resolution, methylation levels are usually averaged over genomic regions in the downstream bioinformatic analysis. Results We demonstrate that on the genome level a single CpG methylation can serve as a more accurate predictor of gene expression than an average promoter / gene body methylation. We define CpG traffic lights (CpG TL) as CpG dinucleotides with a significant correlation between methylation and expression of a gene nearby. CpG TL are enriched in all regulatory regions. Among all promoters, CpG TL are especially enriched in poised ones, suggesting involvement of DNA methylation in their regulation. Yet, binding of only a handful of transcription factors, such as NRF1, ETS, STAT and IRF-family members, could be regulated by direct methylation of transcription factor binding sites (TFBS) or its close proximity. For the majority of TF, an alternative scenario is more likely: methylation and inactivation of the whole regulatory element indirectly represses functional TF binding with a CpG TL being a reliable marker of such inactivation. Conclusions CpG TL provide a promising insight into mechanisms of enhancer activity and gene regulation linking methylation of single CpG to gene expression. CpG TL methylation can be used as reliable markers of enhancer activity and gene expression in applications, e.g. in clinic where measuring DNA methylation is easier compared to directly measuring gene expression due to more stable nature of DNA. Electronic supplementary material The online version of this article (10.1186/s12864-018-5387-1) contains supplementary material, which is available to authorized users.

Atriplex mollis Desf. (Amaranthaceae), a North African endemic halophytic species, is further described in this study. Phylogenetic analysis based on a combined dataset of ITS and ETS rDNA and atpB-rbcL and trnK cpDNA showed that A. mollis is closely related to the Malta- and Gozo-endemic Cremnophyton lanfrancoi Brullo & Pavone. Given this close phylogenetic relationship, A. mollis is also considered among the oldest species of Atriplex, together with C. lanfrancoi. Molecular data also suggest that A. mollis in North Africa, C. lanfrancoi on Malta Island, and Atriplex cana Ledeb. in Eurasian semideserts constitute a separate clade within the tribe Atripliceae. As an 18-month-old shrub, A. mollis can reach a mean height of 44.06 ± 8.09 cm with a leaf area around 1.24 ± 0.15 cm2, and can produce seeds in order of 113.08 ± 28.52 g plant−1. The anatomy of A. mollis shows the presence of male and female developed flowers. Hermaphroditic flowers that may lead to the appearance of male flowers with underdeveloped female organs were rarely found. Three main shapes of ovule (campylotropous, amphitropous, and orthotropous) were found in A. mollis., This study was supported by the ERANETMED2-72-303 PACTORES project financed by the Ministry of Higher Education and Scientific Research of Tunisia and the University of Malta (Argotti Botanic Gardens).

Summary: More than 50% of prostate tumors have a chromosomal rearrangement resulting in aberrant expression of an oncogenic ETS family transcription factor. However, mechanisms that differentiate the function of oncogenic ETS factors expressed in prostate tumors from non-oncogenic ETS factors expressed in normal prostate are unknown. Here, we find that four oncogenic ETS (ERG, ETV1, ETV4, and ETV5), and no other ETS, interact with the Ewing’s sarcoma breakpoint protein, EWS. This EWS interaction was necessary and sufficient for oncogenic ETS functions including gene activation, cell migration, clonogenic survival, and transformation. Significantly, the EWS interacting region of ERG has no homology with that of ETV1, ETV4, and ETV5. Therefore, this finding may explain how divergent ETS factors have a common oncogenic function. Strikingly, EWS is fused to various ETS factors by the chromosome translocations that cause Ewing’s sarcoma. Therefore, these findings link oncogenic ETS function in both prostate cancer and Ewing’s sarcoma. : A subset of ETS transcription factors is oncogenic in prostate. Kedage et al. show that oncogenic ETS, but not other ETS, interact with EWS, and this interaction is necessary for oncogenic functions. Because EWS is fused to ETS factors in Ewing’s sarcoma, this finding links the mechanisms of these diseases. Keywords: prostate cancer, ETS, EWS, Ewing’s sarcoma, ERG

Premise of the study The genus Zanthoxylum in the Rutaceae family of trees and shrubs has a long history of domestication and cultivation in Asia for both economic and medicinal purposes. However, many Zanthoxylum species are morphologically similar and are easily confused. This often leads to false authentication of source materials and confusion in herbal markets, hindering their safe utilization and genetic resource conservation. DNA barcoding is a promising tool for identifying plant taxa. Methods We used three candidate DNA barcoding regions (ITS2, ETS, and trnH-psbA) to identify 69 accessions representing 13 Chinese Zanthoxylum species. The discriminatory capabilities of these regions were evaluated in terms of PCR amplification success, intra- and interspecific divergence, DNA barcoding gaps, and identification efficiency using the BLAST and tree-building methods. Results ITS2 proved the most useful for discriminating Chinese Zanthoxylum species, with a correct identification rate of 100%, and this region also exhibited significantly higher intra- and interspecific divergence. Discussion Phylogenetic analysis confirmed that ITS2 has a powerful discriminatory ability both at and below the species level. We confirmed that ITS2 is a powerful barcoding region for identifying Chinese Zanthoxylum species, and will be useful for analyzing and managing Chinese Zanthoxylum germplasm collections.

Made available in DSpace on 2018-11-26T17:44:56Z (GMT). No. of bitstreams: 0 Previous issue date: 2018-02-01 Portuguese Science Foundation (FCT) through CESAM Portuguese Science Foundation (FCT) Environmental hypercapnia in shallow coastal marine ecosystems can be exacerbated by increasing levels of atmospheric CO2. In these ecosystems organisms are expected to become increasingly subjected to pCO(2) levels several times higher than those inhabiting ocean waters (e.g.: 10,000 mu atm), but still our current understanding on different species capacity to respond to such levels of hypercapnia is limited. Oysters are among the most important foundation species inhabiting these coastal ecosystems, although natural oyster banks are increasingly threatened worldwide. In the present study we studied the effects of hypercapnia on two important oyster species, the pacific oyster C. gigas and the mangrove oyster C. brasiliana, to bring new insights on different species response mechanisms towards three hypercapnic levels (ca. 1,000; 4,000; 10,000 mu atm), by study of a set of biomarkers related to metabolic potential (electron transport system - ETS), antioxidant capacity (SOD, CAT, GSH), cellular damage (LPO) and energetic fitness (GLY), in two life stages (juvenile and adult) after 28 days of exposure. Results showed marked differences between each species tolerance capacity to hypercapnia, with contrasting metabolic readjustment strategies (ETS), different antioxidant response capacities (SOD, CAT, GSH), which generally allowed to prevent increased cellular damage (LPO) and energetic impairment (GLY) in both species. Juveniles were more responsive to hypercapnia stress in both congeners, and are likely to be most sensitive to extreme hypercapnia in the environment. Juvenile C. gigas presented more pronounced biochemical alterations at intermediate hypercapnia (4,000 mu atm) than C. brasiliana. Adult C. gigas showed biochemical alterations mostly in response to high hypercapnia (10,000 mu atm), while adult C. brasiliana were less responsive to this environmental stressor, despite presenting decreased metabolic potential. Our data bring new insights on the biochemical performance of two important oyster species, and suggest that the duration of extreme hypercapnia events in the ecosystem may pose increased challenges for these organisms as their tolerance capacity may be time limited. Univ Aveiro, Dept Biol, Campus Univ Santiago, P-3810193 Aveiro, Portugal Univ Aveiro, CESAM, Campus Univ Santiago, P-3810193 Aveiro, Portugal Univ Estadual Paulista, Inst Biociencias, Campus Litoral Paulista, BR-11330900 Sao Paulo, Brazil Univ Estadual Paulista, Inst Biociencias, Campus Litoral Paulista, BR-11330900 Sao Paulo, Brazil Portuguese Science Foundation (FCT) through CESAM: UID/AMB/50017/2013 Portuguese Science Foundation (FCT): SFRH/BPD/92258/2013 : SFRH/BD/93107/2013

To achieve proper spatiotemporal control of gene expression, transcription factors cooperatively assemble onto specific DNA sequences. The ETS domain protein monomer of GABPα and the B-ZIP domain protein dimer of CREB1 cooperatively bind DNA only when the ETS (C/GCGGAAGT) and CRE (GTGACGTCAC) motifs overlap precisely, producing the ETS↔CRE motif (C/GCGGAAGTGACGTCAC). We designed a Protein Binding Microarray (PBM) with 60-bp DNAs containing four identical sectors, each with 177,440 features that explore the cooperative interactions between GABPα and CREB1 upon binding the ETS↔CRE motif. The DNA sequences include all 15-mers of the form C/GCGGA—–CG—, the ETS↔CRE motif, and all single nucleotide polymorphisms (SNPs), and occurrences in the human and mouse genomes. CREB1 enhanced GABPα binding to the canonical ETS↔CRE motif CCGGAAGT two-fold, and up to 23-fold for several SNPs at the beginning and end of the ETS motif, which is suggestive of two separate and distinct allosteric mechanisms of cooperative binding. We show that the ETS-CRE array data can be used to identify regions likely cooperatively bound by GABPα and CREB1 in vivo, and demonstrate their ability to identify human genetic variants that might inhibit cooperative binding.

Intrinsic resistance and RTK-RAS-MAPK pathway reactivation has limited the effectiveness of MEK and RAF inhibitors (MAPKi) in RAS- and RAF-mutant cancers. To identify genes that modulate sensitivity to MAPKi, we performed genome-scale CRISPR-Cas9 loss-of-function screens in two KRAS mutant pancreatic cancer cell lines treated with the MEK1/2 inhibitor trametinib. Loss of CIC, a transcriptional repressor of ETV1, ETV4, and ETV5, promoted survival in the setting of MAPKi in cancer cells derived from several lineages. ATXN1L deletion, which reduces CIC protein, or ectopic expression of ETV1, ETV4, or ETV5 also modulated sensitivity to trametinib. ATXN1L expression inversely correlates with response to MAPKi inhibition in clinical studies. These observations identify the ATXN1L-CIC-ETS transcription factor axis as a mediator of resistance to MAPKi.

To explore the biosystematics of the Calligonum mongolicum complex (Polygonaceae), the flowering phenological period, breeding and pollination characters and seed set of the complex (Calligonum Mongolicum Turze, Calligonum chinense A. Los., Calligonum gobicum A. Los., Calligonum pumilum A. Los. and Calligonum zaidamense A. Los.) were documented in the Turpan Eremophyte Botanical Garden, China. The sequences of the nuclear ribosomal ITS and ETS region were employed to differentiate the Calligonum mongolicum complex and other species in sect. Medusae. The results showed species of the Calligonum mongolicum complex occupied overlapping flowering periods and had consistent pollination agents. Their breeding systems are all self-compatible, tend to be out-crossing and they interbreed amongst each other (out-crossing index, OCI = 4).The crosses within and amongst species had high seed sets (44 - 65%). Phylogenetic analyses of Calligonum sect. Medusae and the network analysis of nrDNA (ITS and ETS) in the complex suggest interbreeding amongst "species" within the complex and provide evidence for taxonomically merging the five species in the complex. The detected hybridisation, occurring within the complex, suggests the need to improve traditional methods of ex situ plant conservation in botanical gardens for maintaining genetic diversity of Calligonum within and amongst species from different geographic areas.

25 p., figuras, tablas -- Postprint del artículo publicado en Taxon. Versión revisada y corregida -- Contiene material suplementario, Multiple factors related to complex geomorphological and climatic history, in addition to biological factors such as hybridization, hinder the definition of some Mediterranean plant groups. The existence of controversial taxonomic treatments, the possible hybridization events involved, and its unknown evolutionary history, make the Helichrysum pendulum complex of H. sect. Stoechadina an ideal model to understand general processes of Mediterranean plant systematics and evolution. The mosaic range of the complex, which is distributed over several islands and continental areas in the western-central Mediterranean Basin, provides an opportunity to investigate how past connections and disconnections between landmasses may have determined the current geographic distribution of genetic variation in this area. The cpDNA region rpl32-trnL intergenic spacer and the nrDNA region ETS were sequenced for 1–8 individuals from each of the 44 populations sampled, covering all taxa and the whole geographic range of the complex. These individuals were analysed together with a broad sampling of the remaining members of sect. Stoechadina. In addition, detailed multivariate analyses of morphological characters were performed for the whole section and for the H. pendulum complex. Considering together distinctive genetic and morphological traits, our species concept is presented and discussed in a context of integrative taxonomy, and five species are recognized within the complex: H. errerae, H. melitense, H. pendulum, H. saxatile and H. valentinum. The first three species are recognizable by qualitative and quantitative morphological traits, and are genetically distinguishable from the rest as shown by the molecular markers analysed. The two last species are reported here to have a putative ancient hybrid origin and are also genetically distinguishable from the rest but morphologically recognisable only by quantitative characters. Phylogenetic relationships shown by nuclear and chloroplast markers, and an intermediate morphology between the two putative parental taxa, point to H. pendulum and H. italicum as the putative parental taxa for H. saxatile, and H. pendulum and H. stoechas as putative parental taxa for H. valentinum. In a discriminant analysis of the five species, 97.8% of all individuals were classified correctly. The high level of haplotype and ribotype diversity observed in North Africa indicates that this region is either the area of origin of the complex or a secondary contact zone. Our results suggest that the complex colonized several islands and migrated through the Gibraltar and Sicilian Straits during phases of low sea level, favoured by local dispersal events that promoted its gradual range expansion. The occurrence of the complex in the Balearic Islands, which have remained isolated even during low sea level phases, could be explained by stochastic long-distance dispersal events., Financial support from the Spanish Ministerio de Ciencia e Inovación CGL2007-60781/BOS, CGL2009-13322-C03-03/BOS, CGL2010-18631/BOS) and the Catalan government (‘Ajuts a grups consolidats’ 2009/SGR/00439 and 2014/SGR/514) is also acknowledged.

The Spanish Ministry of Science and Technology supported this research through the project CGL2012-38744 and CGL2016- 77401-P to M. Luceño (Principal Investigator), E. Maguilla, M. Escudero, S. Martín-Bravo, and T. Villaverde and an FPU fellowship (AP2012-2189) from the Spanish Government (Ministry of Education, Culture, and Sport) and a Synthesys grant (GB-TAF-2523) by the European Community Research Infrastructure Program to E.Maguilla.

Support was provided by TUBITAK BIDEB project grants to UD and BE; TUBITAK grants no 107S064 and 214Z278 to IK. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. Pea3 transcription factor belongs to the PEA3 subfamily within the ETS domain transcription factor superfamily, and has been largely studied in relation to its role in breast cancer metastasis. Nonetheless, Pea3 plays a role not only in breast tumor, but also in other tissues with branching morphogenesis, including kidneys, blood vasculature, bronchi and the developing nervous system. Identification of Pea3 target promoters in these systems are important for a thorough understanding of how Pea3 functions. Present study particularly focuses on the identification of novel neuronal targets of Pea3 in a combinatorial approach, through curation, computational analysis and microarray studies in a neuronal model system, SH-SY5Y neuroblastoma cells. We not only show that quite a number of genes in cancer, immune system and cell cycle pathways, among many others, are either up- or down-regulated by Pea3, but also identify novel targets including ephrins and ephrin receptors, semaphorins, cell adhesion molecules, as well as metalloproteases such as kallikreins, to be among potential target promoters in neuronal systems. Our overall results indicate that rather than early stages of neurite extension and axonal guidance, Pea3 is more involved in target identification and synaptic maturation. TUBITAK BIDEB TUBITAK -107S064 / 214Z278

The Lucilia group sensu Anderberg and Freire comprises nine South American genera: Belloa, Berroa, Chevreulia, Cuatrecasasiella, Facelis, Gamochaetopsis, Jalcophila, Lucilia and Luciliocline. The aims of this contribution were, using DNA sequences from plastid (rpl32-trnL, trnL-F) and nuclear (ITS and ETS) markers, together with morphological characters, to test the monophyly of the Lucilia group and provide new insight into generic circumscriptions. Our studies, including a broad taxon sampling of Gnaphalieae species, suggest that the Lucilia group is paraphyletic, since Antennaria, Chionolaena, Gamochaeta, Loricaria, Micropsis, Mniodes and Stuckertiella are all nested within the Lucilia group. Morphology and molecular analyses combined showed that the traditional generic circumscription of most of the genera (e.g., Berroa, Chevreulia, Chionolaena, Cuatrecasasiella, Facelis, Jalcophila and Micropsis) correlates with the inferred phylogenetic relationships. Conversely, Lucilia and Luciliocline are non-monophyletic. Lucilia is nested in a clade with Berroa, Facelis and Micropsis. Luciliocline is strongly embedded within the clade Belloa pp + Mniodes. Our results are consistent with Dillon’s study that considered Belloa as a montotypic genus (B. chilensis). Luciliocline and the remaining species of Belloa are accommodated in the genus Mniodes, and the necessary combinations are proposed for the expanded Mniodes. All the analyses showed that the monotypic genera Stuckertiella and Gamochaetopsis are in a well-supported clade nested within Gamochaeta, which implies that taxonomic changes are required also for these genera. Internal relationships in the group and the key morphological characters used in the taxonomy of the group, as well as incongruences found between morphological and molecular analyses, are discussed. Fil: Freire, Susana Edith. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Botánica Darwinion. Academia Nacional de Ciencias Exactas, Físicas y Naturales. Instituto de Botánica Darwinion; Argentina Fil: Chemisquy, Maria Amelia. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Museo Argentino de Ciencias Naturales "Bernardino Rivadavia"; Argentina Fil: Anderberg, Arne A.. Swedish Musem of Natural History; Suecia Fil: Beck, Stephan G.. Universidad Mayor de San Andrés; Bolivia Fil: Meneses, Rosa I.. Universidad Mayor de San Andrés; Bolivia Fil: Leouille, Benoît. Universidade de Sao Paulo; Brasil Fil: Urtubey, Estrella. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Botánica Darwinion. Academia Nacional de Ciencias Exactas, Físicas y Naturales. Instituto de Botánica Darwinion; Argentina

Gamochaeta (tribe Gnaphalieae, Asteraceae) is composed of ca. 60 species primarily distributed in tropical and subtropical America. Within the tribe Gnaphalieae, the genus is characterized by capitula arranged in spikes or head-like clusters, few hermaphroditic central florets, truncate style branches with apical sweeping trichomes, pappus bristles connate at the base into a ring falling as a unit, and achenes with globose twin trichomes. Previous molecular phylogenetic studies have suggested the paraphyly of the genus, but have not provided a basis for redefining generic limits due to incomplete taxon sampling. To address this problem, DNA sequences from the plastid (trnL-F) and nuclear (ETS and ITS) genomes were analyzed from a broad taxon sample representing the full range of morphological variation known in the genus. Our results affirm that Gamochaeta is paraphyletic as presently circumscribed. Two clades can be recognized: one clade that includes the majority of the species currently assigned to Gamochaeta and a second clade that includes Gamochaetopsis, Stuckertiella and seven species of Gamochaeta. We present here a new circumscription of Gamochaeta, including two new combinations, Gamochaeta alpina and Gamochaeta peregrina, and the resurrection of Gamochaeta capitata. Our results also show Omalotheca supina, O. norvegica and O. sylvatica, which were placed by some authors in Gamochaeta or in Gnaphalium, form a monophyletic group distantly related to both genera., Facultad de Ciencias Agrarias y Forestales

Graphical abstract Highlights ► Phylogenetic relationships are reconstructed within Myrceugenia. ► Genus Myrceugenia is monophyletic only when M. fernandeziana is excluded. ► Chilean and Brazilian species are two separate lineages. ► Brazilian species are included in a derived monophyletic group., Myrceugenia is a genus endemic to South America with a disjunct distribution: 12 species occurring mainly in central Chile and approximately 25 in southeastern Brazil. Relationships are reconstructed within Myrceugenia from four plastid markers (partial trnK-matK, rpl32-trnL, trnQ-5′rps16 and rpl16) and two ribosomal nuclear regions (ETS and ITS) using maximum parsimony and Bayesian analyses. Relationships inferred previously from morphological data are not completely consistent with those from molecular data. All molecular analyses support the hypothesis that Myrceugenia is monophyletic, except for M. fernadeziana that falls outside the genus. Chilean species and Brazilian species form two separate lineages. Chilean species form three early diverging clades, whereas Brazilian species are a strongly supported monophyletic group in a terminal position. Least average evolutionary divergence, low resolution, short branches, and high species diversity found in the Brazilian clade suggest rapid radiation. Geographical distributions and phylogenetic reconstructions suggest that extant Myrceugenia species arose in northern Chile followed by colonization southward and finally to the Juan Fernández Islands and southeastern Brazil.

12 pages, 8 figures, 5 tables, Respiratory oxygen consumption is the result of a cell's biochemistry. It is caused by enzymatic activity of the respiratory electron transfer system (ETS). However, in spite of this understanding, respiration models continue to be based on allometric equations relating respiration to body size, body surface, or biomass. The Metabolic Theory of Ecology (MTE) is a current example. It is based on Kleiber's law relating respiration (R) and biomass (M) in the form, View the MathML source, where C is a constant, Ea is the Arrhenius activation energy, k is the Boltzmann constant for an atom or molecule, and T is the temperature in Kelvin. This law holds because biomass packages the ETS. In contrast, we bypass biomass and model respiration directly from its causal relationship with the ETS activity, R = f (ETS). We use a biochemical Enzyme Kinetic Model (EKM) of respiratory oxygen consumption based on the substrate control of the ETS. It postulates that the upper limit of R is set by the maximum velocity, Vmax, of complex I of the ETS and the temperature, and that the substrate availability, S, modulates R between zero and this upper limit. Kinetics of this thermal-substrate regulation is described by the Arrhenius and Michaelis–Menten equations. The EKM equation takes the form View the MathML source where Rg is the molar gas constant and K is the Michaelis–Menten constant. Here, we apply the EKM and the MTE to predict a respiration time-profile throughout the exponential, steady state, and nutrient-limited phases of the marine bacteria Pseudomonas nautica and Vibrio natriegens in acetate-based cultures. Both models were tested by comparing their output with the measured RO2 time-profile. The MTE predicted respiration accurately only in the exponential growth phase, but not during the nutrient limitation part of the stationary phase. In contrast, the EKM worked well throughout both physiological phases as long as the modeled substrates fall with the declining carbon source. Results support the theoretical bases of the EKM. We conclude that the EKM holds promise for predicting respiration at the different physiological states and time-scales important to microbiological studies, Financial support was provided by the Universidad de Las Palmas de Gran Canaria (ULPGC), the Spanish Ministry of Education and Science, the Graduate Program in Oceanography at the ULPGC, ICM-CSIC, and the research grants MODIVUS (CTM2005-04795/MAR), EXOME (CTM 2008-01616), and OITHONA (CTM2007-60052). T. Packard was supported by contract EXMAR SE-539 10/17 (Proyecto Estructurante en Ciencias Marinas). This is contribution #200906 from the Bigelow Laboratory for Ocean Sciences

19 p., Fot.col.; Supplementary Material: Figures S1–S4 are available in the free Electronic Supplement to the online version of this article (http://www.ingentaconnect.com/content/iapt/tax)., [EN] The Filago group (Asteraceae, Gnaphalieae) comprises eleven genera, mainly distributed in Eurasia, northern Africa and northern America: Ancistrocarphus, Bombycilaena, Chamaepus, Cymbolaena, Evacidium, Evax, Filago, Logfia, Micropus, Psilocarphus and Stylocline. The main morphological character that defines the group is that the receptacular paleae subtend, and more or less enclose, the female florets. The aims of this work are, with the use of three chloroplast DNA regions (rpl32-trnL intergenic spacer, trnL intron, and trnL-trnF intergenic spacer) and two nuclear DNA regions (ITS, ETS), to test whether the Filago group is monophyletic; to place its members within Gnaphalieae using a broad sampling of the tribe; and to investigate in detail the phylogenetic relationships among the Old World members of the Filago group and provide some new insight into the generic circumscription and infrageneric classification based on natural entities. Our results do not show statistical support for a monophyletic Filago group. The traditional generic circumscription of most of the genera, as well as the traditional infrageneric classification of the genus Filago, do not correlate with the inferred phylogenetic relationships. A monophyletic circumscription of Filago and a new subgeneric treatment for this genus are proposed, this involving description of a new subgenus (Filago subg. Crocidion Andrés-Sánchez & Galbany, subg. nov.) and four new combinations (Filago subg. Pseudevax (DC.) Andrés-Sánchez & Galbany, comb. et stat. nov.; Filago discolor (DC.) Andrés-Sánchez & Galbany, comb. nov.; Filago gaditana (Pau) Andrés-Sánchez & Galbany, comb. et stat. nov. and Filago griffithii (A. Gray) Andrés-Sánchez & Galbany, comb. nov.). The genera Cymbolaena, Evacidium and Evax are synonymised under Filago. Several incongruences found between chloroplast and nuclear DNA sequence analyses, as well as a notable degree of intraspecific sequence variation in all regions sequenced are documented and discussed.

Multiple gene disorders have been shown to be involved in carcinogenesis. Mutation, translocation and amplification have been identified in so-called oncogenes, and inactivation of antioncogenes by mutation and deletion has been shown. E1AF is an ets-oncogene family transcription factor, and has been shown to upregulate multiple matrix metalloproteinase (MMP) genes that contribute to the malignant phenotype of cancer cells by inducing invasive and metastatic activities. EWS/ETS fusions are frequently observed in Ewing's sarcoma, and we have revealed that EWS/ETS chimeric protein activates telomerase activity by upregulating human telomerase reverse transcriptase (hTERT), but the transcriptional activation of hTERT by EWS/ETS was indirect, and EWS/ETS was seen to function as a co-activator for TERT transcription. A number of oncogenes and cancer-related genes contain AU-rich element (ARE) in non-coding regions of transcribed mRNA. HuR is a RNA-binding protein that has the potential to stabilize ARE-containing mRNAs. HuR is known to shuttle between the nucleus and the cytoplasm via several export pathways. When normal cells are exposed to stress, HuR is exported to the cytoplasm in a chromosome maintenance region 1 (CRM1)-dependent manner. However, we demonstrate that HuR is CRM-1 independently exported to the cytoplasm in oral cancer cells. ARE-mRNAs were also exported to the cytoplasm and stabilized in the oral cancer cells, which were inhibited by HuR knockdown. These findings suggest that transcriptional and translational abnormalities of oncogenes may contribute to the carcinogenesis of oral epithelial cells.

11 pages, 3 figures, 2 tables, The vertical distribution (0–550 m) of zooplankton biomass, and indices of respiration (electron transfer system [ETS]) and structural growth (aminoacyltRNA synthetases activity [AARS]), were studied in waters off the Antarctic Peninsula during the austral summer of 2000. The dominant species were the copepod Metridia gerlachei and the euphausiid Euphausia superba. We observed a vertical krill/copepod substitution in the water column. The zooplankton biomass in the layer at a depth of 200–500 m was of the same magnitude as the biomass in the layer at a depth of 0–200 m, indicating that biomass in the mesopelagic zone is an important fraction of the total zooplankton in Antarctic waters. The metabolic rates of the zooplankton community were sustained by less than 0.5% of the primary production in the area, suggesting that microplankton or small copepods are the main food source. Neither food availability nor predation seemed to control mesozooplankton biomass. The wide time lag between the abundance peak of the dominant copepod (M. gerlachei) and the phytoplankton bloom is suggested to be the main explanation for the low summer zooplankton biomass observed in these waters, We thank the crew of the RV BIO Hespérides, the technicians of the Unidad de Tecnología Marina (CSIC) and the participants of the ESEPAC 2000 cruise for their support at sea, especially S. Agustí for inviting us to participate in the ESEPAC project (ANT97-0273). We are indebted to N. Agawin for kindly supplying primary production values obtained during the cruise, and X. Morán and I.A. Catalán for providing additional data. This work was undertaken while L. Yebra was at the Biological Oceanography Laboratory of the Universidad de Las Palmas de Gran Canaria, supported by grants from the Spanish Science and Education Ministry (MAR97-1036) and the University of Las Palmas de Gran Canaria. Completion of this work was funded by the European Social Fund (I3P programme, CSIC)

It has recently been shown that the majority of prostate cancers harbour a chromosomal rearrangement that fuses the gene for an androgen-regulated prostate-specific serine protease, TMPRSS2, with a member of the ETS family of transcription factors, most commonly ERG. These are among the most common genetic alterations in any human solid tumour. This knowledge may provide us with clues to prostate carcinogenesis, and may lead to the development of important molecular-based biomarkers for patients with localised prostate cancer. The most common variant is fusion between the 5'-untranslated region of TMPRSS2 and the 3' region of ERG. However, over 20 other fusion variants have now been described (involving over 10 different genes) and the number of variants continues to grow. Fusion products can be identified by several techniques, including FISH, RT-PCR, and expression profiling using exon arrays. The protein products associated with the fusion transcripts have not been characterised, and the phenotypic expression of the various products of gene fusion on prostate cancer histology, or on the clinical course of cancer, are not yet understood. Several early cohort studies suggest that the presence of the TMPRSS2:ERG fusion product is associated with relatively poor cancer-specific survival. Studies that examine how individual variants and their associated phenotypes affect prostate cancer presentation and progression are required.

Extrinsic fibroblast growth factor (FGF) signal and intrinsic factors that determine the response of the signal-receiving blastomeres to FGF regulate mesoderm patterning in embryos of the ascidian Halocynthia roretzi. To investigate how cells integrate information from extrinsic and intrinsic inputs, we examined Brachyury (Hr-Bra) promoter activity in the early embryo. Hr-Bra, which encodes a key transcription factor for notochord development, is expressed exclusively in notochord precursors in a manner dependent on the FGF-MEK-MAPK-Ets signaling pathway and on the intrinsic factors Zic and FoxA. Reporter gene expression driven by the 900-bp upstream region of the Hr-Bra promoter was detected as early as the 110-cell stage in notochord precursors by in situ hybridization with a LacZ probe. Deletion analysis combined with MEK inhibitor treatment demonstrated that the − 598/− 499 region carries FGF-responsiveness. Electrophoretic mobility shift assay identified three Ets-binding sites in this region that were required for promoter activity. Further deletion analysis conducted by injecting eggs with reporter constructs at higher concentration suggested that the − 398/− 289 region also has enhancer activity, although ectopic reporter expression was detected in nerve cord and endoderm precursors. The − 398/− 289 region has a Zic-binding site that was also essential for the enhancer activity. These results indicate that Ets- and Zic-binding sites are critical for the initiation of Hr-Bra expression. In conclusion, information from both extrinsic and intrinsic factors is integrated at the level of enhancer of the target gene by direct binding of the transcription factors to the enhancer region.

International audience; Various solid tumors including lung or gastric carcinomas display aberrant activation of the Met receptor which correlates with aggressive phenotypes and poor prognosis. Although downstream signaling of Met is well described, its integration at the transcriptional level is poorly understood. We demonstrate here that in cancer cells harboring met gene amplification, inhibition of Met activity with tyrosine kinase inhibitors or specific siRNA drastically decreased expression of ETV1, ETV4 and ETV5, three transcription factors constituting the PEA3 subgroup of the ETS family, while expression of the other members of the family were less or not affected. Similar link between Met activity and PEA3 factors expression was found in lung cancer cells displaying resistance to EGFR targeted therapy involving met gene amplification. Using silencing experiments, we demonstrate that the PEA3 factors are required for efficient migration and invasion mediated by Met, while other biological responses such as proliferation or unanchored growth remain unaffected. PEA3 overexpression or silencing revealed that they participated in the regulation of the MMP2 target gene involved in extracellular matrix remodeling. Our results demonstrated that PEA3-subgroup transcription factors are key players of the Met signaling integration involved in regulation of migration and invasiveness.

13 pages, 6 figures, 1 table, Respiratory metabolism was compared between 2 different physiological states of acetate- and pyruvate-grown cultures of Pseudomonas nautica and Vibrio natriegens. Here, we analyze 35 h and 520 h experiments in which time-courses of protein, pyruvate, acetate, respiratory CO2 production (RCO2), respiratory O2 consumption (RO2), isocitrate dehydrogenase (IDH) activity, and potential respiration (Φ) were measured. Respiratory quotients (RQs) were calculated as the ratio of the respiration rates (RCO2/RO2). Such RQs are widely used in ocean ecosystem models, in calculations of carbon flux, and in evaluations of the ocean’s metabolic balance. In all the cultures, the RQ tended to increase. In the case of P. nautica on acetate, the RQ rose nearly an order of magnitude from values below 1 during carbon-substrate sufficiency to values close to 10 during carbon-substrate deficiency. In all the cultures, the respiration rates during the growth period paralleled the biomass increase, but after the substrates were exhausted, the respiration rates fell. In contrast, through this same transition period, the IDH activity and the Φ remained relatively high for the first 10 h of carbon-substrate deprivation, and then, these enzyme activities fell slowly, along with the biomass, as the carbon-substrate deprivation continued. The nutritional state of the bacteria affected the RQ, rendering the RQ variable for physiological and ecological purposes. These results argue that ecosystem models, oceanographic calculations of carbon flux, and evaluations of the ocean’s metabolic balance that are influenced by bacterial metabolism need to be reconsidered in light of RQ variability, This work was supported by BIOMBA project CTM 2012-32729/MAR granted to M.G., the University Foundation of Las Palmas by the program ‘Innova Canarias 2020’ financed by Endesa granted to V.R.-K., and a CIE Canarias tri-Continental Atlantic Campus (#CEI10/00018) contract to T.T.P.

The aim of this study was to give results about position of Lamyropsis microcephala within an already existing phylogeny of Lamyropsis genus on the Mediterranean area, by analyzing external transcribed spacer/internal transcribed spacer sequence data. The four Lamyropsis species considered in the model can be recognized as distinct taxa originated from successive migration/speciation events.

As an accompanying manuscript to the release of the honey bee genome, we report the entire sequence of the nuclear (18S, 5.8S, 28S and 5S) and mitochondrial (12S and 16S) ribosomal RNA (rRNA)-encoding gene sequences (rDNA) and related internally and externally transcribed spacer regions of Apis mellifera (Insecta: Hymenoptera: Apocrita). Additionally, we predict secondary structures for the mature rRNA molecules based on comparative sequence analyses with other arthropod taxa and reference to recently published crystal structures of the ribosome. In general, the structures of honey bee rRNAs are in agreement with previously predicted rRNA models from other arthropods in core regions of the rRNA, with little additional expansion in non-conserved regions. Our multiple sequence alignments are made available on several public databases and provide a preliminary establishment of a global structural model of all rRNAs from the insects. Additionally, we provide conserved stretches of sequences flanking the rDNA cistrons that comprise the externally transcribed spacer regions (ETS) and part of the intergenic spacer region (IGS), including several repetitive motifs. Finally, we report the occurrence of retrotransposition in the nuclear large subunit rDNA, as R2 elements are present in the usual insertion points found in other arthropods. Interestingly, functional R1 elements usually present in the genomes of insects were not detected in the honey bee rRNA genes. The reverse transcriptase products of the R2 elements are deduced from their putative open reading frames and structurally aligned with those from another hymenopteran insect, the jewel wasp Nasonia (Pteromalidae). Stretches of conserved amino acids shared between Apis and Nasonia are illustrated and serve as potential sites for primer design, as target amplicons within these R2 elements may serve as novel phylogenetic markers for Hymenoptera. Given the impending completion of the sequencing of the Nasonia genome, we expect our report eventually to shed light on the evolution of the hymenopteran genome within higher insects, particularly regarding the relative maintenance of conserved rDNA genes, related variable spacer regions and retrotransposable elements.

The ID1 protein, an inhibitor of basic helix–loop–helix transcription factors, has been involved in multiple cellular processes including cell cycle regulation, apoptosis, and angiogenesis. To evaluate the importance of ID1 in malignant melanoma, tumour cell expression was examined by immunohistochemistry in 119 cases of nodular melanoma using tissue microarray technique, and related to multiple tumour markers including proliferation, p16 expression, angiogenesis and patient survival. Strong ID1 expression was significantly associated with increased tumour thickness, and significantly reduced survival. Also, increased ID1 was associated with loss of thrombospondin-1 (TSP-1) expression, a known inhibitor of angiogenesis, and increased intensity of ephrin-A1 and its receptor EPHA2. Presence of BRAF mutations was related to strong ID1 expression, but there was no relationship with p16 protein expression. Further, no significant correlation was found between ID1 and microvessel density. In conclusion, our study supports a significant role of the ID1 protein in melanoma progression and patient prognosis. The absence of correlation with p16 protein expression and angiogenesis suggests that other regulatory pathways and mechanisms might be influenced by ID1 in melanomas. An inverse relation between ID1 and TSP-1 expression support an important role of ID1 in the regulation of this complex multitarget protein.

Although the functions of Hox genes in anterior–posterior patterning and their clustered organization are well conserved among metazoans, some Hox genes have lost their original function, as exemplified by zen, ftz and bicoid in Drosophila. The Hox2 gene of amphioxus has also lost its original function and instead is expressed specifically in the preoral pit. As new cis-elements governing its expression in the preoral pit must have been essential for retention of AmphiHox2, we analyzed the transcriptional regulation of AmphiHox2. Although it is possible to make transgenic amphioxus, several technical limitations restrict their practical use; thus, we analyzed the cis-regulatory region surrounding AmphiHox2 in transgenic ascidians (Ciona intestinalis). We found that Ets binding sites of AmphiHox2 functioned in the ascidian embryo. As the amphioxus Ets1/2 homologue is expressed in the preoral pit, we concluded that AmphiHox2 is activated by Ets1/2 in the preoral pit. These analyses demonstrate the utility of Ciona embryos as a transgenic system for analyses of cis-elements from animals whose embryos are relatively inaccessible, such as amphioxus and hemichordates.

Myeloid Elf-1 like factor (MEF) is an ETS protein, which activates the promoters of granulocyte macrophage colony-stimulating factor, interleukin-3, lysozyme, human beta defensin-2 and perforin. In spite of its many known functions, little is known about MEF transcriptional regulation. Here, we cloned the 5′-flanking region of human MEF gene and identified a TATA-less promoter region at −204/−54 which contains 4 putative binding sites for Sp1, two of which are essential in up-regulating MEF activity. These were proven by EMSA and blocking Sp1 using RNAi or mithramycin A treatment of HEK293 cells. Our results suggest that Sp1 constitutively regulates the MEF gene.

Erm, a member of the PEA3 group within the Ets family of transcription factors, is expressed in murine and human lymphocytes. Here, we show that in the human Molt4 lymphoblastic cell line, the erm gene expression is regulated by the conventional PKC (cPKC) pathway. To better characterize the molecular mechanism by which cPKC regulates Erm transcription in Molt4 cells, we tested proximal promoter deletions of the human gene, and identified a specific cPKC-regulated region between positions −420 and −115 upstream of the first exon.

The p16INK4a–RB pathway plays a critical role in preventing inappropriate cell proliferation and is often targeted by viral oncoproteins during immortalization. Latent membrane protein 1 (LMP1) of Epstein-Barr virus (EBV) is often present in EBV-associated proliferative diseases and is critical for the immortalizing and transforming activity of EBV. Unlike other DNA tumor virus oncoproteins, which possess immortalizing activity, LMP1 does not bind to retinoblastoma tumor suppressor protein, but instead blocks the expression of p16INK4a tumor suppressor gene. However, it has been unclear how LMP1 represses the p16INK4a gene expression. Here, we report that LMP1 promotes the CRM1-dependent nuclear export of Ets2, which is an important transcription factor for p16INK4a gene expression, thereby reducing the level of p16INK4a expression. We further demonstrate that LMP1 also blocks the function of E2F4 and E2F5 (E2F4/5) transcription factors through promoting their nuclear export in a CRM1-dependent manner. As E2F4/5 are essential downstream mediators for a p16INK4a-induced cell cycle arrest, these results indicate that the action of LMP1 on nuclear export has two effects on the p16INK4a–RB pathway: (1) repression of p16INK4a expression and (2) blocking the downstream mediator of the p16INK4a–RB pathway. These results reveal a novel activity of LMP1 and increase an understanding of how viral oncoproteins perturb the p16INK4a–RB pathway.

The ascidian Ciona intestinalis is a marine invertebrate belonging to the sister group of the vertebrates, the tunicates. Its compact genome and simple, experimentally tractable embryos make Ciona well-suited for the study of cell-fate specification in chordates. Tunicate larvae possess a characteristic chordate body plan, and many developmental pathways are conserved between tunicates and vertebrates. Previous studies have shown that FGF signals are essential for neural induction and patterning at sequential steps of Ciona embryogenesis. Here we show that two different ETS family transcription factors, Ets1/2 and Elk1/3/4, have partially redundant activities in the anterior neural plate of gastrulating embryos. Whereas Ets1/2 promotes pigment cell formation in lateral lineages, both Ets1/2 and Elk1/3/4 are involved in the activation of Myt1L in medial lineages and the restriction of Six3/6 expression to the anterior-most regions of the neural tube. We also provide evidence that photoreceptor cells arise from posterior regions of the presumptive sensory vesicle, and do not depend on FGF signaling. Cells previously identified as photoreceptor progenitors instead form ependymal cells and neurons of the larval brain. Our results extend recent findings on FGF-dependent patterning of anterior–posterior compartments in the Ciona central nervous system.