Project description:Strigolactones (SLs) are plant hormones involved in plant growth, stress response and plant-organism interactions. DWARF14 (D14) is a α/β hydrolase involved in the perception of SLs in plants. Zaxinone has been reported as a plant endogenous apocarotenoid that modulates growth and biotic interactions, however, the molecular basis of its perception remains elusive. Here we identified a new D14 ligand, the apocarotenoid zaxinone. We showed that hundreds of genes, and specifically CAROTENOID CLEAVAGE DEOXYGENASE7 (CCD7) and CCD8, require At/OsD14, and at lower extent MORE AXILLARY BRANCHING2 (MAX2)/D3, for their transcriptional induction. We find that zaxinone and D14 physically interact in vitro with lower affinity than GR24; the resolution of the AtD14-zaxinone complex, by X-ray crystallography, provided additional evidence that zaxinone antagonizes SL by binding D14. Zaxinone treatments in d14 mutants revealed that At/OsD14 are essential to trigger the induction of CCD7 and CCD8 genes, SL content (roots and exudates) and Striga seed germination validating their interaction in planta. Moreover, OsD14 is required to trigger the zaxinone growth-promoting effect in roots. These findings reveal a perception mechanism for the hormone candidate zaxinone through the SL receptor D14 to regulate transcription of SL genes, SL content and growth in higher plants.
Project description:Larix kaempferi is important afforestation species in northeastern China and is one of the most drought-tolerant pine species.Larix kaempferi has a very complex genetic background and a large genome, and study on the molecular mechanisms of drought resistance is still lag behind. Four-month-old of L. kaempferi seedlings grown in the greenhouse into two groups, the drought treatment group was no watering for 14 d (D14), while normally watered seedlings were used as control group (CK). All plants harvested on the same day and at the same time for the upper part of seedling (circa 3 cm from the top), frozen in liquid nitrogen. We designed six samples (CK-1, CK-2, CK-3, D14-1, D14-2, and D14-3) in total for Illumina HiSeq.
Project description:We performed mRNA 3'end sequencing experiments on three biological replicates of HeLa cells depleted of MATR3, PTBP1/2, controls, or combined depletion of MATR3/PTBP1/2. Cells were fractionated into cytoplasmic and nuclear RNAn and only the nuclear RNA was used. Library preparation was done with the QuantSeq library kit (Lexogen) according to manufacturer’s recommendations. Replicates 1 and 2 were prepared with the QuantSeq forward library kit, replicates 3 and 4 with the QuantSeq reverse library kit. All libraries were sequenced on Illumina HiSeq2 machines in a single-end manner with a read length of 100 nt.
Project description:Gene expressions of murine germinal center and naive B cells on Affymetrix platform The experiment include 3 d14 GC B1-8, 3 d14 GC V23 and 4 NaM-CM-/ve samples
Project description:Mechanical force is a fundamental regulator of bone development and homeostasis. Mechanosensitive osteocytes are the most abundant bone cells that form interconnected dendrites to respond to mechanical stimuli and interact with the bone-forming osteoblasts and the bone-remodeling osteoclasts. However, the molecular mechanisms underlying osteocyte maturation and dendrite formation remain unclear. By generating a Piezo1 "knock-out" osteocyte cell line, we identified a key role of Piezo1-mediated mechanotransduction in osteocyte differentiation. QRT-PCR analysis revealed delayed osteocyte differentiation in the Piezo1 KO cells relative to WT cells. By performing bulk RNA sequencing of WT and Piezo1 KO OCY454 cells at early (D1), intermediate (D14), and late (D28) stages of differentiation allowed for the identification of key signaling pathways in driving normal osteocyte dfiferentiation, as well as those regulated by Piezo1 mechanotransduction.
Project description:The F-box protein MORE AXILLARY GROWTH 2 (MAX2) is a central component in the signaling cascade of strigolactones (SLs) as well as of the smoke derived karrikins (KARs) and the so far unknown endogenous KAI2 ligand (KL). The two groups of molecules are involved in overlapping and unique developmental processes, and signal-specific outcomes are attributed to perception by the paralogous α/β-hydrolases DWARF14 (D14) for SL and KARRIKIN INSENSITIVE 2/ HYPOSENSITIVE TO LIGHT (KAI2/HTL) for KAR/KL. Additionally, depending on which receptor is activated, specific members of the SUPPRESSOR OF MAX2 1 (SMAX1) – LIKE (SMXL) 6, 7, 8 clade control KAR/KL and SL responses respectively. As proteins that function in the same signal transduction pathway often occur in large protein complexes, we aimed at discovering new players of the MAX2, D14 and KAI2 protein network by tandem affinity purification using Arabidopsis cell cultures. When using MAX2 as a bait, various proteins were co-purified among which general components of the Skp1-Cullin-F-box complex and members of the CONSTITUTIVE PHOTOMORPHOGENIC 9 signalosome. Here, we report the identification of a novel interactor of MAX2, a type 5 serine/threonine protein phosphatase, designated PHYTOCHROME-ASSOCIATED PROTEIN PHOSPHATASE 5 (PAPP5). Quantitative affinity purification pointed at PAPP5 as being more present in KAI2 rather than D14 protein complexes. In agreement, mutant analysis suggests that PAPP5 modulates KAR/KL-dependent seed germination in suboptimal conditions and seedling development. Additionally, PAPP5 was found to dephosphorylate MAX2 in vivo independent of the synthetic strigolactone analog, rac-GR24. Together, by analyzing the protein complexes to which MAX2, D14 and KAI2 belong, we revealed a new MAX2 interactor that might act through dephosphorylating MAX2 to control mainly KAR/KL signaling.
Project description:Comparative small RNA seq analysis of WT and global p73KO Mouse Tracheal Epithelial Cell (MTECs) during the course of their differentiation (Air-Liquid Interface ALI D0, D4, D7, D14) aimed to determine the role of p73 in motile multiciliogenesis.
Project description:Purpose: To understand the role of SUMOylation of c-Fos in the differential regulation of target genes and altered cellular pathways upon STm infection. To address this, we performed a RNA-seq experiment with stable over expressing WT-FOS or SUMO-def-FOS in f10 c-FOS-Knock out MEFs upon STm infection. Methods: Three biological replicates of c-FOS-KO-WT-FOS, c-FOS-KO-WT-FOS with salmonella infection, c-FOS-KO-SUMO-def-FOS and c-FOS-KO-SUMO-def-FOS with salmonella infection, cells were collected and total RNA was extracted according to kit's protocol. The total RNA was used to generate a cDNA library using Quantseq 3' mRNA kit. Results: Transcriptional profiling revealed that genes involved in immune response, proliferation, metastasis etc. are differentially regulated in salmonella infected c-FOS-KO-SUMO-def-FOS MEFs compare to salmonella infected c-FOS-KO-WT-FOS MEFs. Conclusions: Our study revealed the extensive transcriptomics analysis from c-FOS-KO-WT-FOS and c-FOS-KO-SUMO-def-FOS MEFs upon salmonella infection. We found that SUMOylation of c-Fos provides selectivity that causes differential regulation of target genes which are involved in immune response, proliferation etc. pathways of host. Together, our findings illuminate an important regulatory role played by SUMOylated c-Fos upon STm infection.