Sestrin-2 (SESN2) as a stress-metabolic protein is known for its anti-oxidative effects as a downstream factor of PERK pathways in mammalian cells. However, the expression patterns of SESN2 in conjunction with the UPR signaling against to ER stress on porcine oocyte maturation in vitro, have not been reported. Therefore, we confirmed the expression pattern of SESN2 protein, for which to examine the relationship between PERK signaling and SESN2 in porcine oocyte during IVM. We investigated the SESN2 expression patterns using Western blot analysis in denuded oocytes (DOs), cumulus cells (CCs), and cumulus-oocyte complexes (COCs) at 22 and 44 h of IVM. As expected, the SESN2 protein level significantly increased (p < 0.01) in porcine COCs during 44 h of IVM. We investigated the meiotic maturation after applying ER stress inhibitor in various concentration (50, 100 and 200 μM) of tauroursodeoxycholic acid (TUDCA). We confirmed significant increase (p < 0.05) of meiotic maturation rate in TUDCA 200 μM treated COCs for 44 h of IVM. Finally, we confirmed the protein level of SESN2 and meiotic maturation via regulating ER-stress by only tunicamycin (Tm), only TUDCA, and Tm + TUDCA treatment in porcine COCs. As a result, treatment of the TUDCA following Tm pre-treatment reduced SESN2 protein level in porcine COCs. In addition, SESN2 protein level significantly reduced in only TUDCA treated porcine COCs. Our results suggest that the SESN2 expression is related to the stress mediator response to ER stress through the PERK signaling pathways in porcine oocyte maturation.
Periodontal ligament (PDL) tissue is a connective tissue that is interposed between the roots of the teeth and the inner wall of the alveolar bone socket. PDL is always exposed to physiologic mechanical force such as masticatory force and PDL cells play important roles during orthodontic tooth movement by synthesizing and secreting different mediators involved in bone remodeling. The Wnt/β-catenin signaling pathway was recently shown to play a significant role in the control of bone formation. In the present study, we applied cyclic tensile stress of 20% elongation to cultured human PDL cells and assessed its impact after six days upon components of the Wnt/β-catenin signaling pathway. RTPCR analysis showed that Wnt1a, Wnt3a, Wnt10b and the Wnt receptor LRP5 were down-regulated, whereas the Wnt inhibitor DKK1 was up-regulated in response to these stress conditions. In contrast, little change was detected in the mRNA expression of Wnt5a, Wnt7b, Fz1, and LRP6. By western blotting we found decreased expression of the β-catenin and p-GSK-3β proteins. Our results thus show that mechanical stress suppresses the canonical Wnt/β-catenin signaling pathway in PDL cells.
Several E3 ubiquitin ligases have been associated with the response to abiotic and biotic stresses in higher plants. Here, we report that the hot pepper (Capsicumannuum) abscisic acid (ABA)-InsensitiveRINGprotein1gene(CaAIR1) is essential for a hypersensitive response to drought stress. CaAIR1 contains a C3HC4-type RING finger motif, which plays a role for attachment of ubiquitins to the target protein, and a putative transmembrane domain. The expression levels of CaAIR1 are upregulated in pepper leaves by ABA treatments, drought, and NaCl, suggesting its role in the response to abiotic stress. Our analysis showed that CaAIR1 displays self-ubiquitination and localized in the nucleus. We generated CaAIR1-silenced peppers via virus-induced gene silencing (VIGS) and CaAIR1-overexpressing (OX) transgenic Arabidopsis plants to evaluate their responses to ABA and drought. VIGS of CaAIR1 in pepper plants conferred an enhanced tolerance to drought stress, which was accompanied by low levels of transpirational water loss in the drought-treated leaves. CaAIR1-OX plants displayed an impaired sensitivity to ABA during seed germination, seedling, and adult stages. Moreover, these plants showed enhanced sensitivity to drought stress because of reduced stomatal closure and decreased expression of stress-responsive genes. Thus, our data indicate that CaAIR1 is a negative regulator of the ABA-mediated drought-stress tolerance mechanism.
Drought and high salinity are the most important abiotic factors limiting plant development, growth, and crop productivity in agriculture (Munns and Tester 2008, Sengupta and Majumder 2009, Zhu 2002). As sessile organisms, plants are frequently exposed to drought and high salinity conditions, which alter water potential and cause osmotic stress, leading to serious damage to plant tissues (Bartels and Sunkar 2005, Boudsocq and Lauriere 2005). During exposure to water stress, plants display many physiological changes, such as reduction of water content, closure of stomata, and decreased cell enlargement and growth. In addition, severe and continuous water stress in plants causes the cessation of photosynthesis and disturbance of metabolism, and finally results in death (Nath et al. 2005, Shao et al. 2008). To adapt to these abiotic stress conditions, plants show a variety of responses, including the accumulation of abscisic acid (ABA) and expression of a large number of stress-related proteins (Krasensky and Jonak 2012, Lee and Luan 2012, Skriver and Mundy 1990, Stewart and Lee 1974). Although the cellular and molecular responses to environmental stress are well studied (Hasegawa et al. 2000, Thomashow 1999), the mechanisms underlying the functional modifications caused by osmotic stress are yet to be clarified, because of the complexity at the cellular level as well as at the whole plant level (Ashraf and Harris 2004, Flowers 2004, Foolad et al. 2003a, 2003b, Xiong et al. 2002).
Recently there are many reports that signaling pathways of abiotic stress and biotic stress are correlated. These relations are not only antagonistic but also synergistic. In this project we are searching the common components in abiotic and biotic stress signaling through proteome and transcriptome analysis. In this project, we are profiling the transcriptome under ABA and biotic stress treatment and searching the common genes which were regulated in both treatment. Furthermore, we are analyzing the secretome and proteome induced under C.maydis. It would be expected that integrative analysis of transcriptome and proteome will presents us the candidate genes to develop abiotic/biotic stress tolerant transgenic plants.