CRISPR-Mediated Endogenous Activation of Fibroin Heavy Chain Gene Triggers Cellular Stress Responses in Bombyx mori Embryonic Cells

The silkworm Bombyx mori is an economically important insect, as it is the main producer of silk. Fibroin heavy chain (FibH) gene, encoding the core component of silk protein, is specifically and highly expressed in silk gland cells but not in the other cells. Although the silkworm FibH gene has been well studied in transcriptional regulation, its biological functions in the development of silk gland cells remain elusive. In this study, we constructed a CRISPRa system to activate the endogenous transcription of FibH in Bombyx mori embryonic (BmE) cells, and the mRNA expression of FibH was successfully activated. In addition, we found that FibH expression was increased to a maximum at 60 h after transient transfection of sgRNA/dCas9-VPR at a molar ratio of 9:1. The qRT-PCR analysis showed that the expression levels of cellular stress response-related genes were significantly up-regulated along with activated FibH gene. Moreover, the lyso-tracker red and monodansylcadaverine (MDC) staining assays revealed an apparent appearance of autophagy in FibH-activated BmE cells. Therefore, we conclude that the activation of FibH gene leads to up-regulation of cellular stress responses-related genes in BmE cells, which is essential for understanding silk gland development and the fibroin secretion process in B. mori.

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Flavonoids: Potential Candidates for the Treatment of Neurodegenerative Disorders

Biomedicines ◽

10.3390/biomedicines9020099 ◽

2021 ◽

Vol 9 (2) ◽

pp. 99

Author(s):

Shweta Devi ◽

Vijay Kumar ◽

Sandeep Kumar Singh ◽

Ashish Kant Dubey ◽

Jong-Joo Kim

Keyword(s):

Stress Response ◽

Stress Responses ◽

Neurodegenerative Disorders ◽

Cell Damage ◽

Cellular Stress ◽

Clinical Manifestations ◽

Cellular Stress Response ◽

Dramatic Rise ◽

Cellular Stress Responses

Neurodegenerative disorders, such as Parkinson’s disease (PD), Alzheimer’s disease (AD), Amyotrophic lateral sclerosis (ALS), and Huntington’s disease (HD), are the most concerning disorders due to the lack of effective therapy and dramatic rise in affected cases. Although these disorders have diverse clinical manifestations, they all share a common cellular stress response. These cellular stress responses including neuroinflammation, oxidative stress, proteotoxicity, and endoplasmic reticulum (ER)-stress, which combats with stress conditions. Environmental stress/toxicity weakened the cellular stress response which results in cell damage. Small molecules, such as flavonoids, could reduce cellular stress and have gained much attention in recent years. Evidence has shown the potential use of flavonoids in several ways, such as antioxidants, anti-inflammatory, and anti-apoptotic, yet their mechanism is still elusive. This review provides an insight into the potential role of flavonoids against cellular stress response that prevent the pathogenesis of neurodegenerative disorders.

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Cellular Stress Responses in Radiotherapy

Cells ◽

10.3390/cells8091105 ◽

2019 ◽

Vol 8 (9) ◽

pp. 1105 ◽

Cited By ~ 26

Author(s):

Kim ◽

Lee ◽

Seo ◽

Kim ◽

...

Keyword(s):

Stress Responses ◽

Molecular Mechanisms ◽

Cellular Stress ◽

Treatment Strategies ◽

Cellular Stress Response ◽

Cytotoxic Effects ◽

Cancer Cell Death ◽

Damage Repair ◽

Cellular Stress Responses ◽

Radiation Induced

Radiotherapy is one of the major cancer treatment strategies. Exposure to penetrating radiation causes cellular stress, directly or indirectly, due to the generation of reactive oxygen species, DNA damage, and subcellular organelle damage and autophagy. These radiation-induced damage responses cooperatively contribute to cancer cell death, but paradoxically, radiotherapy also causes the activation of damage-repair and survival signaling to alleviate radiation-induced cytotoxic effects in a small percentage of cancer cells, and these activations are responsible for tumor radio-resistance. The present study describes the molecular mechanisms responsible for radiation-induced cellular stress response and radioresistance, and the therapeutic approaches used to overcome radioresistance.

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Flavonoids: Potential Candidates for the Treatment of Neuro-Degenerative Disorders

10.20944/preprints202101.0384.v1 ◽

2021 ◽

Author(s):

Shweta Devi ◽

Vijay Kumar ◽

Sandeep Kumar Singh ◽

Ashish Kant Dubey ◽

Jong-Joo Kim

Keyword(s):

Stress Response ◽

Stress Responses ◽

Neurodegenerative Disorders ◽

Cell Damage ◽

Cellular Stress ◽

Clinical Manifestations ◽

Cellular Stress Response ◽

Degenerative Disorders ◽

Dramatic Rise ◽

Cellular Stress Responses

Neurodegenerative disorders such as Parkinson’s disease (PD), Alzheimer’s disease (AD), Amyloidal lateral sclerosis (ALS), and Huntington disease (HD) are the most concerned disorders due to the lack of effective therapeutics and dramatic rise in affected cases. Although these disorders have diverse clinical manifestations, yet they all share a common cellular stress response. These cellular stress responses including neuroinflammation, oxidative stress, proteotoxicity, and ER-stress, which combats with stress conditions, but the overwhelming cellular stress response induces cell damage. Small molecules such as flavonoids could reduce cellular stress and have gained much attention in recent years. Evidence has shown the potential use of flavonoids in several ways such as antioxidants, anti-inflammatory, and anti-apoptotic, yet their mechanism is still elusive. This review provides an insight into the mechanistic ways of flavonoids against cellular stress response that prevent the pathogenesis of neurodegenerative disorders.

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Retrograde signaling and plant stress: plastid signals initiate cellular stress responses

Current Opinion in Plant Biology ◽

10.1016/j.pbi.2008.06.002 ◽

2008 ◽

Vol 11 (5) ◽

pp. 509-513 ◽

Cited By ~ 114

Author(s):

Aurora Piñas Fernández ◽

Åsa Strand

Keyword(s):

Stress Responses ◽

Cellular Stress ◽

Plant Stress ◽

Retrograde Signaling ◽

Cellular Stress Responses

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P25 gene regulation in Bombyx mori silk gland: two promoter-binding factors have distinct tissue and developmental specificities

Molecular and Cellular Biology ◽

10.1128/mcb.12.12.5768-5777.1992 ◽

1992 ◽

Vol 12 (12) ◽

pp. 5768-5777

Author(s):

B Durand ◽

J Drevet ◽

P Couble

Keyword(s):

Bombyx Mori ◽

Head Capsule ◽

Regulatory Elements ◽

Silk Gland ◽

Related Factor ◽

Developmental Studies ◽

Hormonal Change ◽

Specific Expression ◽

Gland Cells ◽

Gene Status

The gene encoding the silk protein P25 is expressed in the posterior silk gland of Bombyx mori with strict territorial and developmental specificities. The cis-acting regulatory elements previously located within the 441-bp 5' proximal sequence of the gene were examined for protein-binding capacities. We identified two factors, BMFA and SGFB, that lead to prominent band shifts and the target sites for which are included in a region homologous to the fibroin gene enhancer sequence. Analysis of the tissue-specific incidence of both factors showed that BMFA is ubiquitous, whereas SGFB is restricted to the silk gland cells. However, SGFB was found in both posterior and middle silk gland cells and therefore likely directs organ-specific, but not territory-specific, expression. Developmental studies throughout the fourth larval molt, at which the P25 gene status changes from derepressed to repressed, revealed that BMFA is reversibly modified at the transition from intermolt to molt. Indeed, the preexisting BMFA is replaced by a structurally related factor, BMFA', during the 2 h following head capsule apolysis. The exact temporal coincidence of this conversion with the onset of gene repression suggests that BMFA' is involved in transcription inactivation and likely results from a transduction process initiated by the hormonal change at molting.

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Cellular Stress Responses: A Balancing Act

Molecular Cell ◽

10.1016/j.molcel.2010.10.008 ◽

2010 ◽

Vol 40 (2) ◽

pp. 175 ◽

Cited By ~ 8

Author(s):

Feng Chen ◽

Allyson Evans ◽

John Pham ◽

Brian Plosky

Keyword(s):

Stress Responses ◽

Cellular Stress ◽

Cellular Stress Responses

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The Phosphorylation-Dependent Regulation of Mitochondrial Proteins in Stress Responses

Journal of Signal Transduction ◽

10.1155/2012/931215 ◽

2012 ◽

Vol 2012 ◽

pp. 1-12 ◽

Cited By ~ 27

Author(s):

Yusuke Kanamaru ◽

Shiori Sekine ◽

Hidenori Ichijo ◽

Kohsuke Takeda

Keyword(s):

Stress Responses ◽

Map Kinases ◽

Mitochondrial Fission ◽

Cellular Stress ◽

Degradation Process ◽

Mitochondrial Proteins ◽

Phosphoglycerate Mutase ◽

Mitogen Activated Protein ◽

Cellular Stress Responses ◽

Autophagic Degradation

To maintain cellular homeostasis, cells are equipped with precise systems that trigger the appropriate stress responses. Mitochondria not only provide cellular energy but also integrate stress response signaling pathways, including those regulating cell death. Several lines of evidence suggest that the mitochondrial proteins that function in this process, such as Bcl-2 family proteins in apoptosis and phosphoglycerate mutase family member 5 (PGAM5) in necroptosis, are regulated by several kinases. It has also been suggested that the phosphorylation-dependent regulation of mitochondrial fission machinery, dynamin-related protein 1 (Drp1), facilitates appropriate cellular stress responses. However, mitochondria themselves are also damaged by various stresses. To avoid the deleterious effects exerted by damaged mitochondria, cells remove these mitochondria in a selective autophagic degradation process called mitophagy. Interestingly, several kinases, such as PTEN-induced putative kinase 1 (PINK1) in mammals and stress-responsive mitogen-activated protein (MAP) kinases in yeast, have recently been shown to be involved in mitophagy. In this paper, we focus on the phosphorylation-dependent regulation of mitochondrial proteins and discuss the roles of this regulation in the mitochondrial and cellular stress responses.

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