VE-statin/egfl7 Expression in Endothelial Cells Is Regulated by a Distal Enhancer and a Proximal Promoter under the Direct Control of Erg and GATA-2

Angiogenesis is the process by which new blood vessels arise from existing ones by the budding out of endothelial cell capillaries from the luminal side of blood vessels. Blood vessel formation is essential for organ development during embryogenesis and is associated with several physiological and pathological processes, such as wound healing and tumor development. The VE-statin/egfl7 gene is specifically expressed in endothelial cells during embryonic development and in the adult. We studied here the regulatory mechanisms that control this tissue-specific expression. RT-qPCR analyses showed that the specificity of expression of VE-statin/egfl7 in endothelial cells is not shared with its closest neighbor genes notch1 and agpat2 on the mouse chromosome 2. Chromatin-immunoprecipitation analysis of histone modifications at the VE-statin/egfl7 locus showed that the chromatin is specifically opened in endothelial cells, but not in fibroblasts at the transcription start sites. A 13 kb genomic fragment of promoter was cloned and analyzed by gene reporter assays which showed that two conserved regions are important for the specific expression of VE-statin/egfl7 in endothelial cells; a −8409/−7563 enhancer and the −252/+38 region encompassing the exon-1b transcription start site. The latter contains essential GATA and ETS-binding sites, as assessed by linker-scanning analysis and site-directed mutagenesis. An analysis of expression of the ETS and GATA transcription factors showed that Erg, Fli-1 and GATA-2 are the most highly expressed factors in endothelial cells. Erg and GATA-2 directly control the expression of the endogenous VE-statin/egfl7 while Fli-1 probably exerts an indirect control, as assessed by RNA interference and chromatin immunoprecipitation. This first detailed analysis of the mechanisms that govern the expression of the VE-statin/egfl7 gene in endothelial cells pinpoints the specific importance of ETS and GATA factors in the specific regulation of genes in this cell lineage

Rice early flowering1, a CKI, phosphorylates DELLA protein SLR1 to negatively regulate gibberellin signalling

The plant hormone gibberellin (GA) is crucial for multiple aspects of plant growth and development. To study the relevant regulatory mechanisms, we isolated a rice mutant earlier flowering1, el1, which is deficient in a casein kinase I that has critical roles in both plants and animals. el1 had an enhanced GA response, consistent with the suppression of EL1 expression by exogenous GA3. Biochemical characterization showed that EL1 specifically phosphorylates the rice DELLA protein SLR1, proving a direct evidence for SLR1 phosphorylation. Overexpression of SLR1 in wild-type plants caused a severe dwarf phenotype, which was significantly suppressed by EL1 deficiency, indicating the negative effect of SLR1 on GA signalling requires the EL1 function. Further studies showed that the phosphorylation of SLR1 is important for maintaining its activity and stability, and mutation of the candidate phosphorylation site of SLR1 results in the altered GA signalling. This study shows EL1 a novel and key regulator of the GA response and provided important clues on casein kinase I activities in GA signalling and plant development

Seed use in the field: delivering seeds for restoration success

© 2020 The Authors. Restoration Ecology published by Wiley Periodicals LLC on behalf of Society for Ecological Restoration Seed delivery to site is a critical step in seed-based restoration programs. Months or years of seed collection, conditioning, storage, and cultivation can be wasted if seeding operations are not carefully planned, well executed, and draw upon best available knowledge and experience. Although diverse restoration scenarios present different challenges and require different approaches, there are common elements that apply to most ecosystems and regions. A seeding plan sets the timeline and details all operations from site treatments through seed delivery and subsequent monitoring. The plan draws on site evaluation data (e.g. topography, hydrology, climate, soil types, weed pressure, reference site characteristics), the ecology and biology of the seed mix components (e.g. germination requirements, seed morphology) and seed quality information (e.g. seed purity, viability, and dormancy). Plan elements include: (1) Site treatments and seedbed preparation to remove undesirable vegetation, including sources in the soil seed bank; change hydrology and soil properties (e.g. stability, water holding capacity, nutrient status); and create favorable conditions for seed germination and establishment. (2) Seeding requirements to prepare seeds for sowing and determine appropriate seeding dates and rates. (3) Seed delivery techniques and equipment for precision seed delivery, including placement of seeds in germination-promotive microsites at the optimal season for germination and establishment. (4) A monitoring program and adaptive management to document initial emergence, seedling establishment, and plant community development and conduct additional sowing or adaptive management interventions, if warranted. (5) Communication of results to inform future seeding decisions and share knowledge for seed-based ecological restoration