Novel hyperactive transposons for genetic modification of induced pluripotent and adult stem cells: a nonviral paradigm for coaxed differentiation.

Adult stem cells and induced pluripotent stem cells (iPS) hold great promise for regenerative medicine. The development of robust nonviral approaches for stem cell gene transfer would facilitate functional studies and potential clinical applications. We have previously generated hyperactive transposases derived from Sleeping Beauty, using an in vitro molecular evolution and selection paradigm.

Prion protein in Alzheimer's pathogenesis: a hot and controversial issue.

The role for cellular prion protein PrP(c) in beta-amyloid (Abeta) oligomer-induced synaptic impairment is a topic of great interest and some controversy. In this issue of EMBO Molecular Medicine Aguzzi and co-workers explore the contribution of PrP(c) to deficient long term potentiation (LTP) and soluble Abeta levels in an Alzheimer's disease mouse model and show that the role of prions in Abeta related toxicity is far from 'black and white' suggesting complex interpretations of the data available thus far.

The X chromosome in immune functions: when a chromosome makes the difference.

In response to various immune challenges, females show better survival than males; the X chromosome has an important role in this immunological advantage. X chromosome-linked diseases are usually restricted to males, who have only one copy of the X chromosome; however, females are more prone to autoimmune diseases, and the X chromosome may be involved in the breakdown of self tolerance. Several hypotheses have been proposed in recent years that support a role for the X chromosome in shaping autoimmune responses.

The culprit behind amyloid beta peptide related neurotoxicity in Alzheimer's disease: oligomer size or conformation?

Since the reformulation of the amyloid cascade hypothesis to focus on oligomeric aggregates of amyloid beta as the prime toxic species causing Alzheimer's disease, many researchers refocused on detecting a specific molecular assembly of defined size thatis the main trigger of Alzheimer's disease. The result has been the identification of a host of molecular assemblies containing from two up to a hundred molecules of the amyloid beta peptide, which were all found to impair memory formation in mice. This clearly demonstrates that size is insufficient to define toxicity and peptide conformation has to be taken into account.

Trafficking to the outer polar domain defines the root-soil interface.

In animals, the interface between organism and environment is constituted by the epithelium. In plants, the exchange of nutrients and signals between root and soil is crucial for their survival, but the cellular mechanisms underlying the epithelium-like function and specific localization of proteins to the root surface have not been identified. Here we analyze the mechanism of polar delivery to the root-soil interface of the proteins BOR4, ABCG37, and PEN3, which transport nutrients, transport plant hormones, and are required for pathogen defense, respectively.

NINJA connects the co-repressor TOPLESS to jasmonate signalling.

Jasmonoyl-isoleucine (JA-Ile) is a plant hormone that regulates a broad array of plant defence and developmental processes. JA-Ile-responsive gene expression is regulated by the transcriptional activator MYC2 that interacts physically with the jasmonate ZIM-domain (JAZ) repressor proteins. On perception of JA-Ile, JAZ proteins are degraded and JA-Ile-dependent gene expression is activated.

Response of Pseudomonas aeruginosa PAO1 to low shear modelled microgravity involves AlgU regulation.

As a ubiquitous environmental organism that is occasionally part of the human flora, Pseudomonas aeruginosa could pose a health hazard for the immunocompromised astronauts during long-term missions. Therefore, insights into the behaviour of P. aeruginosa under spaceflight conditions were gained using two spaceflight-analogue culture systems: the rotating wall vessel (RWV) and the random position machine (RPM).

Genetic modification of the inner ear lateral semicircular canal phenotype of the Bmp4 haplo-insufficient mouse.

In the mouse, development of the lateral semicircular canal of the inner ear is sensitive to Bmp4 heterozygosity. In the C57BL6 background 30% of the heterozygotes display circling behavior, 66% have a specific defect in the vestibular part of the inner ear, namely the constriction, interruption or absence of the lateral semicircular canal. Only mice having both ears affected display circling behavior.

Hormone-mediated promotion of trichome initiation in plants is conserved but utilizes species- and trichome-specific regulatory mechanisms.

Plant trichome initiation is steered by diverse developmental and environmental cues, through molecular mechanisms that remain elusive in most plant species. Using a robust experimental method to investigate the molecular mechanisms by which phytohormones modulate leaf trichome formation, we verified the effect of jasmonates, cytokinins and gibberellins in Arabidopsis (). All three phytohormones promoted Arabidopsis trichome initiation, but caused divergent effects on trichome maturation and other leaf parameters.

Role of PIN-mediated auxin efflux in apical hook development of Arabidopsis thaliana.

The apical hook of dark-grown Arabidopsis seedlings is a simple structure that develops soon after germination to protect the meristem tissues during emergence through the soil and that opens upon exposure to light. Differential growth at the apical hook proceeds in three sequential steps that are regulated by multiple hormones, principally auxin and ethylene. We show that the progress of the apical hook through these developmental phases depends on the dynamic, asymmetric distribution of auxin, which is regulated by auxin efflux carriers of the PIN family.

The MCM-binding protein ETG1 aids sister chromatid cohesion required for postreplicative homologous recombination repair.

The DNA replication process represents a source of DNA stress that causes potentially spontaneous genome damage. This effect might be strengthened by mutations in crucial replication factors, requiring the activation of DNA damage checkpoints to enable DNA repair before anaphase onset. Here, we demonstrate that depletion of the evolutionarily conserved minichromosome maintenance helicase-binding protein ETG1 of Arabidopsis thaliana resulted in a stringent late G2 cell cycle arrest.

PIN phosphorylation is sufficient to mediate PIN polarity and direct auxin transport.

The plant hormone auxin plays a crucial role in regulating plant development and plant architecture. The directional auxin distribution within tissues depends on PIN transporters that are polarly localized on the plasma membrane. The PIN polarity and the resulting auxin flow directionality are mediated by the antagonistic actions of PINOID kinase and protein phosphatase 2A.

Cardioselective nitric oxide synthase 3 gene transfer protects against myocardial reperfusion injury.

Nitric oxide modulates the severity of myocardial ischemia-reperfusion (I/R) injury. We investigated whether cardioselective nitric oxide synthase 3 (NOS3) gene transfer could confer myocardial protection against I/R injury in pigs and examined potential molecular mechanisms. I/R injury was induced by balloon occlusion of the left anterior descending artery for 45 min followed by 4 or 72 h reperfusion.

The ang3 mutation identified the ribosomal protein gene RPL5B with a role in cell expansion during organ growth.

The role of translation in the regulation of higher plant growth and development is not well understood. Mutational analysis is a powerful tool to identify and study the function of genes related to a biological process, such as growth. Here we analyzed functionally the angusta3 (ang3) narrow leaf mutant.

Receptor endocytosis and signaling in plants.

The emerging complexity of plant endocytic systems puts it on a par with their animal counterparts, reflecting an essential role in signal transduction. The endocytic machinery regulates the space and the time of signal transduction and processing in the cell. Plants possess numerous cell surface receptor-like kinases (RLKs) (more than 600 members in Arabidopsis thaliana and 1100 in rice), a trend attributed to their indeterminate mode of growth, the absence of cell migration, and the need for adaptation towards the environment.

The flowering world: a tale of duplications.

Flowering plants contain many genes, most of which were created during the past 200 or so million years through small- and large-scale duplications. Paleo-polyploidy events, in particular, have been the subject of much recent research. There is a growing consensus that one or more genome doubling or merging events occurred early during the evolution of the flowering plants, and that many lineages have since undergone additional, independent and more recent duplication events.

Proteome-wide substrate analysis indicates substrate exclusion as a mechanism to generate caspase-7 versus caspase-3 specificity.

Caspase-3 and -7 are considered functionally redundant proteases with similar proteolytic specificities. We performed a proteome-wide screen on a mouse macrophage lysate using the N-terminal combined fractional diagonal chromatography technology and identified 46 shared, three caspase-3-specific, and six caspase-7-specific cleavage sites. Further analysis of these cleavage sites and substitution mutation experiments revealed that for certain cleavage sites a lysine at the P5 position contributes to the discrimination between caspase-7 and -3 specificity.

Albinism and cell viability in cycloartenol synthase deficient Arabidopsis.

Phenotypes of Arabidopsis thaliana that carry mutations in CYCLOARTENOL SYNTHASE 1 (CAS1) which is required in sterol biosynthesis have been described. Knockout mutant alleles are responsible of a male-specific transmission defect. Plants carrying a weak mutant allele cas1-1 accumulate 2,3-oxidosqualene, the substrate of CAS1, in all analyzed organs.

Transcriptional control of the cell cycle.

Cell division is a highly coordinated process. In the last decades, many plant cell cycle regulators have been identified. Strikingly, only a few transcriptional regulators are known, although a significant amount of the genome is transcribed in a cell cycle phase-dependent manner.

A survey of TonB-dependent receptors in fluorescent pseudomonads.

For bacteria with an aerobic lifestyle, iron is in the oxidized Fe(3+) form, hence poorly soluble. The solution is the synthesis and excretion of siderophores with a high affinity for iron. These ferrisiderophores are recognized by TonB-dependent outer membrane receptors in Gram-negative bacteria.

The Gram-positive side of plant-microbe interactions.

Plant growth and development are significantly influenced by the presence and activity of microorganisms. To date, the best-studied plant-interacting microbes are Gram-negative bacteria, but many representatives of both the high and low G+C Gram-positives have excellent biocontrol, plant growth-promoting and bioremediation activities. Moreover, actinorhizal symbioses largely contribute to the global biological nitrogen fixation and many Gram-positive bacteria promote other types of symbioses in tripartite interactions.

Clone-based functional genomics.

Annotated genomes have provided a wealth of information about gene structure and gene catalogs in a wide range of species. Taking advantage of these developments, novel techniques have been implemented to investigate systematically diverse aspects of gene and protein functions underpinning biology processes. Here, we review functional genomics applications that require the mass production of cloned sequence repertoires, including ORFeomes and silencing tag collections.

A novel method for tissue-specific RNAi rescue in Drosophila.

Targeted gene silencing by RNA interference allows the study of gene function in plants and animals. In cell culture and small animal models, genetic screens can be performed--even tissue-specifically in Drosophila--with genome-wide RNAi libraries. However, a major problem with the use of RNAi approaches is the unavoidable false-positive error caused by off-target effects.

Emerging potential of transposons for gene therapy and generation of induced pluripotent stem cells.

Effective gene therapy requires robust delivery of the desired genes into the relevant target cells, long-term gene expression, and minimal risks of secondary effects. The development of efficient and safe nonviral vectors would greatly facilitate clinical gene therapy studies. However, nonviral gene transfer approaches typically result in only limited stable gene transfer efficiencies in most primary cells.

High frequency of single-copy T-DNA transformants produced by floral dip in CRE-expressing Arabidopsis plants.

For genetic transformation of plants, floral dip with Agrobacterium often results in integration of multiple T-DNA copies at a single locus and frequently in low and unstable transgene expression. To obtain efficient single-copy T-DNA transformants, two CRE/loxP recombinase-based simplifying strategies for complex T-DNA loci were compared. A T-DNA vector with oppositely oriented loxP sites was transformed into CRE-expressing and wild-type control Arabidopsis thaliana plants.