scholarly journals A SUBTILISIN-LIKE SERINE PROTEASE1 (OsSUBSrP1), plays an important role in the wax and cutin pathway, is essential for panicle development in rice

2021 ◽  
Author(s):  
Asif Ali ◽  
Tingkai Wu ◽  
Hongyu Zhang ◽  
Peizhou Xu ◽  
Syed Adeel Zafar ◽  
...  
Keyword(s):  
Cell Death ◽  
Global Gene Expression ◽  
Blue Staining ◽  
The Novel ◽  
Knock Out ◽  
Panicle Development ◽  
Global Gene Expression Analysis ◽  
Normal Meiosis ◽  
Spikelet Degeneration

Panicle degeneration is a severe physiological defect and causes reduction in grain yield. In this study, we characterized and presented the functional analysis of our previously reported mutant apa1331 (apical panicle abortion1331) that showed apical spikelet degeneration. The anthers from the apical spikelets of apa1331 were degenerated, pollen-less and showed lack of cuticle formation. Transverse sections showed normal meiosis till stage 5-6, however, defects in post-meiotic microspore development were found at stage 8-9 in apa1331. Measurement of wax and cutin analysis showed a significant reduction in anthers of apa1331 compared to Wildtype (WT). Quantification of H2O2 and MDA has indicated the excessive ROS (reactive oxygen species) in apa1331. Trypan blue staining, and TUNEL assay revealed cell death and excessive DNA fragmentation in apa1331. Map-based cloning and Mutmap analysis identified a candidate gene (LOC_Os04g40720) that is a SUBTILISIN-LIKE SERINE PROTEASE (OsSUBSrP1) which harbored an SNP (A>G) in apa1331. CRISPR-mediated knock-out lines of OsSUBSrP1 displayed spikelet degeneration comparable to apa1331. Global gene expression analysis revealed a significant downregulation of wax and cutin biosynthesis genes e.g., OsWDA1, OsMS2 and OsCER4 in apa1331. Our study reports the novel role of SUBSrP1 in ROS-mediated cell death in panicle development.

scholarly journals The Agr-Like Quorum Sensing System Is Required for Pathogenesis of Necrotic Enteritis Caused by Clostridium perfringens in Poultry

2017 ◽  
Vol 85 (6) ◽  
Author(s):  
Qiang Yu ◽  
Dion Lepp ◽  
Iman Mehdizadeh Gohari ◽  
Tao Wu ◽  
Hongzhuan Zhou ◽  
...  
Keyword(s):  
Quorum Sensing ◽  
Clostridium Perfringens ◽  
Global Gene Expression ◽  
Toxin Production ◽  
Phospholipid Metabolism ◽  
Necrotic Enteritis ◽  
Content Type ◽  
Protein Levels ◽  
Global Gene Expression Analysis

ABSTRACT Clostridium perfringens encodes at least two different quorum sensing (QS) systems, the Agr-like and LuxS, and recent studies have highlighted their importance in the regulation of toxin production and virulence. The role of QS in the pathogenesis of necrotic enteritis (NE) in poultry and the regulation of NetB, the key toxin involved, has not yet been investigated. We have generated isogenic agrB-null and complemented strains from parent strain CP1 and demonstrated that the virulence of the agrB-null mutant was strongly attenuated in a chicken NE model system and restored by complementation. The production of NetB, a key NE-associated toxin, was dramatically reduced in the agrB mutant at both the transcriptional and protein levels, though not in a luxS mutant. Transwell assays confirmed that the Agr-like QS system controls NetB production through a diffusible signal. Global gene expression analysis of the agrB mutant identified additional genes modulated by Agr-like QS, including operons related to phospholipid metabolism and adherence, which may also play a role in NE pathogenesis. This study provides the first evidence that the Agr-like QS system is critical for NE pathogenesis and identifies a number of Agr-regulated genes, most notably netB, that are potentially involved in mediating its effects. The Agr-like QS system thus may serve as a target for developing novel interventions to prevent NE in chickens.

scholarly journals The Role of Pseudo-Orthocaspase (SyOC) of Synechocystis sp. PCC 6803 in Attenuating the Effect of Oxidative Stress

2021 ◽  
Vol 12 ◽  
Author(s):  
Saul Lema A ◽  
Marina Klemenčič ◽  
Franziska Völlmy ◽  
Maarten Altelaar ◽  
Christiane Funk
Keyword(s):  
Oxidative Stress ◽  
Cell Death ◽  
Building Blocks ◽  
Amino Acid Residues ◽  
Knock Out ◽  
Functional Homology ◽  
Its Gene ◽  
Pcc 6803

Caspases are proteases, best known for their involvement in the execution of apoptosis—a subtype of programmed cell death, which occurs only in animals. These proteases are composed of two structural building blocks: a proteolytically active p20 domain and a regulatory p10 domain. Although structural homologs appear in representatives of all other organisms, their functional homology, i.e., cell death depending on their proteolytical activity, is still much disputed. Additionally, pseudo-caspases and pseudo-metacaspases, in which the catalytic histidine-cysteine dyad is substituted with non-proteolytic amino acid residues, were shown to be involved in cell death programs. Here, we present the involvement of a pseudo-orthocaspase (SyOC), a prokaryotic caspase-homolog lacking the p10 domain, in oxidative stress in the model cyanobacterium Synechocystis sp. PCC 6803. To study the in vivo impact of this pseudo-protease during oxidative stress its gene expression during exposure to H2O2 was monitored by RT-qPCR. Furthermore, a knock-out mutant lacking the pseudo-orthocaspase gene was designed, and its survival and growth rates were compared to wild type cells as well as its proteome. Deletion of SyOC led to cells with a higher tolerance toward oxidative stress, suggesting that this protein may be involved in a pro-death pathway.

Identification of mutation biomarkers underpinning colon cancer sidedness and cetuximab sensitivity.

2018 ◽  
Vol 36 (4_suppl) ◽  
pp. 629-629
Author(s):  
Timothy Joseph Yeatman ◽  
Mingli Yang ◽  
Michael J. Schell ◽  
Andrey Loboda ◽  
Michael Nebozhyn ◽  
...  
Keyword(s):  
Gene Expression ◽  
Global Gene Expression ◽  
Ras Mutation ◽  
Mutation Status ◽  
Therapeutic Opportunity ◽  
Global Gene Expression Analysis ◽  
Msi Analysis ◽  
Mutant Genes ◽  
Ranking Analysis

629 Background: Currently, extended RAS testing ( KRAS/ NRAS) in colorectal cancer (CRC) patients identifies only non-responders to EGFR inhibitor (EGFRi) therapies, and accurate prediction of drug-sensitive subpopulations remains problematic in patients, even with wild-type RAS. Moreover, the molecular basis for the laterality of anti-EGFR sensitivity is poorly understood. Methods: 468 CRCs were analyzed by global gene expression analysis, DNA sequencing (1321 cancer related genes) and MSI analysis. Tumors were stratified by a validated gene expression cetuximab sensitivity (CTX-S) score, and then correlated with identified high frequency mutations, resulting in a ranking of CTX-S score-associated mutated genes (see Table). Results: Ranking analysis revealed MSI-H status and KRAS mutation as the most negatively-correlated among all patients and MSS patients, respectively. Conversely, APC and TP53 were the most highly positively-correlated mutant genes in both all patient- and MSS-cases. Deeper analysis revealed that the combination of mutant APC + TP53 ( A + P) was more common in left vs. right CRCs (52% vs 21%), and even more pronounced in MSS vs MSI cases (47% vs 2%). CTX-S scores were highest in mutant A + P patients with WT RAS, but surprisingly, were nearly as high in mutant A + P patients with mutant RAS. CRC PDX models validated these results with a favorable CR/PR/SD vs PD association. Conclusions: Here we report the discovery of a cooperative role of APC and TP53 mutations in identifying EGFRi-sensitive CRC subpopulations. Our data suggest that addition of the routine sequencing of APC and TP53 to extended RAS testing may expand the EGFRi therapeutic opportunity (by up to 25%), regardless of RAS mutation status. [Table: see text]

scholarly journals Proapoptotic Bad Involved in Brain Development, When Severely Defected, Induces Dramatic Malformation in Zebrafish

2021 ◽  
Vol 22 (9) ◽  
pp. 4832
Author(s):  
Jo-Chi Hung ◽  
Jen-Leih Wu ◽  
Jiann-Ruey Hong
Keyword(s):  
Cell Death ◽  
Programmed Cell Death ◽  
Environmental Stress ◽  
Brain Development ◽  
The Novel ◽  
Germ Layers ◽  
Cell Death Signaling ◽  
Morpholino Oligonucleotides ◽  
Material Factor

The BH3-only molecule Bad regulates cell death via its differential protein phosphorylation, but very few studies address its effect on early embryonic development in vertebrate systems. In this work, we examined the novel role of zebrafish Bad in the initial programmed cell death (PCD) for brain morphogenesis through reducing environmental stress and cell death signaling. Bad was considered to be a material factor that because of the knockdown of Bad by morpholino oligonucleotides, PCD was increased and the reactive oxygen species (ROS) level was enhanced, which correlated to trigger a p53/caspase-8 involving cell death signaling. This Bad knockdown-mediated environmental stress and enhanced cell dying can delay normal cell migration in the formation of the three germ layers, especially the ectoderm, for further brain development. Furthermore, Bad defects involved in three-germ-layers development at 8 hpf were identified by in situ hybridization approach on cyp26, rtla, and Sox17 pattern expression markers. Finally, the Bad knockdown-induced severely defected brain was examined by tissue section from 24 to 48 h postfertilization (hpf), which correlated to induce dramatic malformation in the hindbrain. Our data suggest that the BH3-only molecule Bad regulates brain development via controlling programmed cell death on overcoming environmental stress for reducing secondary cell death signaling, which suggests that correlates to brain developmental and neurological disorders in this model system.

scholarly journals Treatment of SARS-CoV-2 induced pneumonia with NAD+ in a mouse model

2020 ◽  
Author(s):  
Yisheng Jiang ◽  
Yongqiang Deng ◽  
Tiantian Ma ◽  
Huanhuan Pang ◽  
Zeping Hu ◽  
...  
Keyword(s):  
Cell Death ◽  
Global Gene Expression ◽  
Infection Model ◽  
Nad Metabolism ◽  
Effective Strategies ◽  
Effect Of Treatment ◽  
Global Gene Expression Analysis ◽  
Mouse Study ◽  
Mouse Lungs

Abstract The global COVID-19 epidemic has spread rapidly around the world and has already caused the death of more than one million people. As there is yet no vaccine, it is urgent to develop effective strategies to treat COVID-19 patients. Here, we used a mouse-adapted SARS-CoV-2 infection model to explore potential therapeutic targets for SARS-CoV-2 pneumonia. Global gene expression analysis of infected mouse lungs revealed dysregulation of genes associated with NAD+ metabolism, immune response and cell death, similar to that in COVID-19 patients. We therefore investigated the effect of treatment with NAD+ and found that the pneumonia phenotypes, including excessive inflammatory cell infiltration and embolization in SARS-CoV-2 infected lungs were significantly rescued by boosting NAD+ levels. Most notable, cell death was suppressed substantially (>65%) by NAD+ supplementation. Thus, our in vivo mouse study supports trials for treating COVID-19 patients with NAD+ or its precursors.

scholarly journals Role of brassinosteroids in rice spikelet differentiation and degeneration under soil-drying during panicle development

2019 ◽  
Vol 19 (1) ◽  
Author(s):  
Weiyang Zhang ◽  
Jiayan Sheng ◽  
Yunji Xu ◽  
Fei Xiong ◽  
Yunfei Wu ◽  
...  
Keyword(s):  
Rice Cultivar ◽  
Soil Drying ◽  
Nonstructural Carbohydrates ◽  
Inflorescence Development ◽  
Expression Levels ◽  
Panicle Development ◽  
Drying Conditions ◽  
Rice Panicles ◽  
Spikelet Degeneration

Abstract Background Brassinosteroids (BRs) are a new group of plant hormones and play important roles in plant growth and development. However, little information is available if BRs could regulate spikelet development in rice (Oryza sativa L.) especially under soil-drying conditions. This study investigated whether and how BRs mediate the effect of soil-drying on spikelet differentiation and degeneration in rice. A rice cultivar was field-grown and exposed to three soil moisture treatments during panicle development, that is, well-watered (WW), moderate soil-drying (MD) and severe soil-drying (SD). Results Compared with the WW treatment, the MD treatment enhanced BRs biosynthesis in young panicles, increased spikelet differentiation and reduced spikelet degeneration. The SD treatment had the opposite effects. Changes in expression levels of key rice inflorescence development genes (OsAPO2 and OsTAW1), ascorbic acid (AsA) content, and activities of enzymes involved AsA synthesis and recycle, and amount of nonstructural carbohydrates (NSC) in young panicles were consistent with those in BRs levels, whereas hydrogen peroxide (H2O2) content showed opposite trend. Knockdown of the BRs synthesis gene OsD11 or application of a BRs biosynthesis inhibitor to young panicles markedly decreased OsAPO2 and OsTAW1 expression levels, BRs and AsA contents, activities of enzymes involved AsA synthesis and recycle, NSC amount in rice panicles and spikelet differentiation but increased the H2O2 content and spikelet degeneration compared to the control (the wide type or application of water). The opposite effects were observed when exogenous BRs were applied. Conclusions The results suggest that BRs mediate the effect of soil-drying on spikelet differentiation and degeneration, and elevated BRs levels in rice panicles promote spikelet development under MD by enhancing inflorescence meristem activity, AsA recycle and NSC partitioning to the growing panicles.

scholarly journals Human vtRNA1-1 Levels Modulate Signaling Pathways and Regulate Apoptosis in Human Cancer Cells

Biomolecules ◽  
2020 ◽  
Vol 10 (4) ◽  
pp. 614 ◽  
Author(s):  
Lisamaria Bracher ◽  
Iolanda Ferro ◽  
Carlos Pulido-Quetglas ◽  
Marc-David Ruepp ◽  
Rory Johnson ◽  
...  
Keyword(s):  
Cell Death ◽  
Cell Lines ◽  
Human Cancer ◽  
Apoptosis Resistance ◽  
Protein Coding ◽  
Knock Out ◽  
Human Cancer Cells ◽  
Extrinsic Apoptosis ◽  
A Cell

Regulatory non-protein coding RNAs perform a remarkable variety of complex biological functions. Previously, we demonstrated a role of the human non-coding vault RNA1-1 (vtRNA1-1) in inhibiting intrinsic and extrinsic apoptosis in several cancer cell lines. Yet on the molecular level, the function of the vtRNA1-1 is still not fully clear. Here, we created HeLa knock-out cell lines revealing that prolonged starvation triggers elevated levels of apoptosis in the absence of vtRNA1-1 but not in vtRNA1-3 knock-out cells. Next-generation deep sequencing of the mRNome identified the PI3K/Akt pathway and the ERK1/2 MAPK cascade, two prominent signaling axes, to be misregulated in the absence of vtRNA1-1 during starvation-mediated cell death conditions. Expression of vtRNA1-1 mutants identified a short stretch of 24 nucleotides of the vtRNA1-1 central domain as being essential for successful maintenance of apoptosis resistance. This study describes a cell signaling-dependent contribution of the human vtRNA1-1 to starvation-induced programmed cell death.

scholarly journals Lactate administration increases mRNA expression of PGC-1α and UCP3 in mouse skeletal muscle

2016 ◽  
Vol 41 (6) ◽  
pp. 695-698 ◽  
Author(s):  
Yu Kitaoka ◽  
Kohei Takeda ◽  
Yuki Tamura ◽  
Hideo Hatta
Keyword(s):  
Skeletal Muscle ◽  
Gene Expression Analysis ◽  
Gastrocnemius Muscle ◽  
Potential Role ◽  
Quantitative Polymerase Chain Reaction ◽  
Global Gene Expression ◽  
Signalling Molecule ◽  
Global Gene Expression Analysis ◽  
Polymerase Chain

To examine the potential role of lactate as a signalling molecule in skeletal muscle, we performed global gene expression analysis of the mouse gastrocnemius muscle, 3 h after lactate administration using the Affymetrix GeneChip system (Affymetrix, Santa Clara, Calif., USA). Among the top 15 genes with the largest fold change, increased expression of Ppargc1a, Pdk4, and Ucp3 was confirmed using real-time quantitative polymerase chain reaction. Our findings suggest that lactate serves as a signal for upregulating genes related to mitochondrial function.

Protection of adult rat cardiac myocytes from ischemic cell death: role of caveolar microdomains and δ-opioid receptors

2006 ◽  
Vol 291 (1) ◽  
pp. H344-H350 ◽  
Author(s):  
Hemal H. Patel ◽  
Brian P. Head ◽  
Heidi N. Petersen ◽  
Ingrid R. Niesman ◽  
Diane Huang ◽  
...  
Keyword(s):  
Cell Death ◽  
Cardiac Myocytes ◽  
Opioid Receptors ◽  
Signaling Molecules ◽  
Blue Staining ◽  
Dependent Manner ◽  
Protective Effects ◽  
Cardiac Protection ◽  
Presence And Absence

The role of caveolae, membrane microenvironments enriched in signaling molecules, in myocardial ischemia is poorly defined. In the current study, we used cardiac myocytes prepared from adult rats to test the hypothesis that opioid receptors (OR), which are capable of producing cardiac protection in vivo, promote cardiac protection in cardiac myocytes in a caveolae-dependent manner. We determined protein expression and localization of δ-OR (DOR) using coimmunohistochemistry, caveolar fractionation, and immunoprecipitations. DOR colocalized in fractions with caveolin-3 (Cav-3), a structural component of caveolae in muscle cells, and could be immunoprecipitated by a Cav-3 antibody. Immunohistochemistry confirmed plasma membrane colocalization of DOR with Cav-3. Cardiac myocytes were subjected to simulated ischemia (2 h) or an ischemic preconditioning (IPC) protocol (10 min ischemia, 30 min recovery, 2 h ischemia) in the presence and absence of methyl-β-cyclodextrin (MβCD, 2 mM), which binds cholesterol and disrupts caveolae. We also assessed the cardiac protective effects of SNC-121 (SNC), a selective DOR agonist, on cardiac myocytes with or without MβCD and MβCD preloaded with cholesterol. Ischemia, simulated by mineral oil layering to inhibit gas exchange, promoted cardiac myocyte cell death (trypan blue staining), a response blunted by SNC (37 ± 3 vs. 59 ± 3% dead cells in the presence and absence of 1 μM SNC, respectively, P < 0.01) or by use of the IPC protocol (35 ± 4 vs. 62 ± 3% dead cells, P < 0.01). MβCD treatment, which disrupted caveolae (as detected by electron microscopy), fully attenuated the protective effects of IPC or SNC, resulting in cell death comparable to that of the ischemic group. By contrast, SNC-induced protection was not abrogated in cells incubated with cholesterol-saturated MβCD, which maintained caveolae structure and function. These findings suggest a key role for caveolae, perhaps through enrichment of signaling molecules, in contributing to protection of cardiac myocytes from ischemic damage.

Sign in / Sign up

Export Citation Format

Share Document