Silencing of PTK7 in Colon Cancer Cells: Caspase-10-Dependent Apoptosis via Mitochondrial Pathway

Protein tyrosine kinase-7 (PTK7) is a catalytically inactive receptor tyrosine kinase (RTK). PTK7 is upregulated in many common human cancers, including colon cancer, lung cancer, gastric cancer and acute myeloid leukemia. The reason for this up-regulation is not yet known. To explore the functional role of PTK7, the expression of PTK7 in HCT 116 cells was examined using small interference (siRNA)-mediated gene silencing. Following transfection, the siRNA successfully suppressed PTK7 mRNA and protein expression. Knocking down of PTK7 in HCT 116 cells inhibited cell proliferation compared to control groups and induced apoptosis. Furthermore, this apoptosis was characterized by decreased mitochondrial membrane potential and activation of caspase-9 and -10. Addition of a caspase-10 inhibitor totally blocked this apoptosis, suggesting that caspase-10 may play a critical role in PTK7-knockdown-induced apoptosis, downstream of mitochondria. These observations may indicate a role for PTK7 in cell proliferation and cell apoptosis and may provide a potential therapeutic pathway for the treatment of a variety of cancers

DETORQUEO, QUIRKY, and ZERZAUST Represent Novel Components Involved in Organ Development Mediated by the Receptor-Like Kinase STRUBBELIG in Arabidopsis thaliana

Intercellular signaling plays an important role in controlling cellular behavior in apical meristems and developing organs in plants. One prominent example in Arabidopsis is the regulation of floral organ shape, ovule integument morphogenesis, the cell division plane, and root hair patterning by the leucine-rich repeat receptor-like kinase STRUBBELIG (SUB). Interestingly, kinase activity of SUB is not essential for its in vivo function, indicating that SUB may be an atypical or inactive receptor-like kinase. Since little is known about signaling by atypical receptor-like kinases, we used forward genetics to identify genes that potentially function in SUB-dependent processes and found recessive mutations in three genes that result in a sub-like phenotype. Plants with a defect in DETORQEO (DOQ), QUIRKY (QKY), and ZERZAUST (ZET) show corresponding defects in outer integument development, floral organ shape, and stem twisting. The mutants also show sub-like cellular defects in the floral meristem and in root hair patterning. Thus, SUB, DOQ, QKY, and ZET define the STRUBBELIG-LIKE MUTANT (SLM) class of genes. Molecular cloning of QKY identified a putative transmembrane protein carrying four C2 domains, suggesting that QKY may function in membrane trafficking in a Ca2+-dependent fashion. Morphological analysis of single and all pair-wise double-mutant combinations indicated that SLM genes have overlapping, but also distinct, functions in plant organogenesis. This notion was supported by a systematic comparison of whole-genome transcript profiles during floral development, which molecularly defined common and distinct sets of affected processes in slm mutants. Further analysis indicated that many SLM-responsive genes have functions in cell wall biology, hormone signaling, and various stress responses. Taken together, our data suggest that DOQ, QKY, and ZET contribute to SUB-dependent organogenesis and shed light on the mechanisms, which are dependent on signaling through the atypical receptor-like kinase SUB

Structure-Function Analysis of STRUBBELIG, an Arabidopsis Atypical Receptor-Like Kinase Involved in Tissue Morphogenesis

Tissue morphogenesis in plants requires the coordination of cellular behavior across clonally distinct histogenic layers. The underlying signaling mechanisms are presently being unraveled and are known to include the cell surface leucine-rich repeat receptor-like kinase STRUBBELIG in Arabidopsis. To understand better its mode of action an extensive structure-function analysis of STRUBBELIG was performed. The phenotypes of 20 EMS and T-DNA-induced strubbelig alleles were assessed and homology modeling was applied to rationalize their possible effects on STRUBBELIG protein structure. The analysis was complemented by phenotypic, cell biological, and pharmacological investigations of a strubbelig null allele carrying genomic rescue constructs encoding fusions between various mutated STRUBBELIG proteins and GFP. The results indicate that STRUBBELIG accepts quite some sequence variation, reveal the biological importance for the STRUBBELIG N-capping domain, and reinforce the notion that kinase activity is not essential for its function in vivo. Furthermore, individual protein domains of STRUBBELIG cannot be related to specific STRUBBELIG-dependent biological processes suggesting that process specificity is mediated by factors acting together with or downstream of STRUBBELIG. In addition, the evidence indicates that biogenesis of a functional STRUBBELIG receptor is subject to endoplasmic reticulum-mediated quality control, and that an MG132-sensitive process regulates its stability. Finally, STRUBBELIG and the receptor-like kinase gene ERECTA interact synergistically in the control of internode length. The data provide genetic and molecular insight into how STRUBBELIG regulates intercellular communication in tissue morphogenesis

Investigation of the contribution from peripheral GnRH-like immunoreactive 'neuroblasts' to the regenerating central nervous system in the protochordate Ciona intestinalis

The neural ganglion of the ascidian Ciona intestinalis regenerates in its entirety within a few weeks of ablation. Here we investigate the role of gonadotropin-releasing hormone-like immunoreactive (GnRH-li) cells in regeneration. Immunocytochemical studies show that in addition to a previously described plexus of GnRH-like neurones located in association with the dorsal strand, the normal adult brain contains GnRH-li neurones. These are predominantly localized to the ventral cortical rind at the posterior of the ganglion. Following ablation, non-process bearing GnRH-li cells appear in the regenerating area within two days. By day 8 post-ablation, process bearing GnRH-li cells are detected close to the regenerating brain. The number of these cells increases at later stages. GnRH-li cells are first detected within the regenerating brain at 14 days post-ablation and their number subsequently increases. These cells are initially concentrated along the ventral surface of the regenerating brain near to the dorsal strand. Double labelling studies with bromodeoxyuridine show that none of the GnRH-li cells are labelled at any stage of regeneration. The data are consistent with a sub-population of the new neurones being derived from GnRH-li neuroblasts born prior to ablation, which migrate from the dorsal strand complex into the regenerating ganglion

Linking structure and species richness to support forest biodiversity monitoring at large scales

Abstract Key message Authors have analyzed the possible correlation between measurements/indicators of forest structure and species richness of many taxonomic or functional groups over three regions of Germany. Results show the potential to use structural attributes as a surrogate for species richness of most of the analyzed taxonomic and functional groups. This information can be transferred to large-scale forest inventories to support biodiversity monitoring. Context We are currently facing a dramatic loss in biodiversity worldwide and this initiated many monitoring programs aiming at documenting further trends. However, monitoring species diversity directly is very resource demanding, in particular in highly diverse forest ecosystems. Aims We investigated whether variables applied in an index of stand structural diversity, which was developed based on forest attributes assessed in the German National Forest Inventory, can be calibrated against richness of forest-dwelling species within a wide range of taxonomic and functional groups. Methods We used information on forest structure and species richness that has been comprehensively assessed on 150 forest plots of the German biodiversity exploratories project, comprising a large range of management intensities in three regions. We tested, whether the forest structure index calculated for these forest plots well correlate with the number of species across 29 taxonomic and functional groups, assuming that the structural attributes applied in the index represent their habitat requirements. Results The strength of correlations between the structural variables applied in the index and number of species within taxonomic or functional groups was highly variable. For some groups such as Aves, Formicidae or vascular plants, structural variables had a high explanatory power for species richness across forest types. Species richness in other taxonomic and functional groups (e.g., soil and root-associated fungi) was not explained by individual structural attributes of the index. Results indicate that some taxonomic and functional groups depend on a high structural diversity, whereas others seem to be insensitive to it or even prefer structurally poor stands. Conclusion Therefore, combinations of forest stands with different degrees of structural diversity most likely optimize taxonomic diversity at the landscape level. Our results can support biodiversity monitoring through quantification of forest structure in large-scale forest inventories. Changes in structural variables over inventory periods can indicate changes in habitat quality for individual taxonomic groups and thus points towards national forest inventories being an effective tool to detect unintended effects of changes in forest management on biodiversity