Gut CaVP is an innate immune protein against bacterial challenge in amphioxus Branchiostoma belcheri

Zhuang, Zhenhong; Zhao, Xianliang; Li, Hui; Wang, Sanying; Peng, Xuan‐xian

doi:10.1016/j.fsi.2011.05.004

Cited by 28 publications

(11 citation statements)

References 32 publications

(24 reference statements)

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“…The membranes were rinsed three times for 10 min with TNT buffer and incubated with goat anti-mouse HRP-conjuaged IgG at a dilution of 1∶4000 in TNT buffer containing 5% skim milk for 1 h at room temperature. The membranes were developed with substrate (ECL, Electrochemiluminescence) until optimum color developed [23].…”

Section: Methodsmentioning

confidence: 99%

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Study on the Apoptosis Mechanism Induced by T-2 Toxin

et al. 2013

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T-2 toxin is known to induce apoptosis in mammalian cells. The mechanism of apoptosis induced by T-2 toxin has been proposed to be linked with oxidative stress and mitochondrial pathway. In the current study, the toxic effect of T-2 on Hela, Bel-7402, and Chang liver cells was examined in dose-dependent and time-dependent manner by MTT assay. Caspase-3 was found to be up-regulated under T-2 toxin stress, which suggested that T-2 toxin induced cell apoptosis. Endogenous GSH and MDA levels in all three cell lines were found down- and up-regulated respectively, which indicated the link between toxic effect of T-2 toxin and intracellular oxidative stress. It was also found by MTT assay that NAC, which maintained the level of GSH in cells, could protect cells from death. Western-blot result showed that the level of both activated Caspase-8 and Caspase-9 increased when cells were treated by T-2 toxin. Caspase-9 was found to be activated earlier than Caspase-8. It was also found that p53 was up-regulated under T-2 toxin stress in the study. These results implied that the effect of T-2 toxin on cells was apoptosis rather than necrosis, and it was probably induced through mitochondrial pathway. To the best of our knowledge, the present study is the first to show that JunD is down-regulated in T-2 toxin induced apoptosis. By construction of an over-expression vector for the JunD gene, we observed that the survival ratio of JunD over-expressed cells obviously increased under T-2 toxin stress. These results suggested that the mechanism of T-2 induced cell death was closely connected with oxidative stress, and that JunD plays an important role in the defensive process against T-2 toxin stress.

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Section: Methodsmentioning

confidence: 99%

“…After screening by G418, the JunD expression level was detected by Western-blot analysis with anti-JunD antibody as the first antibody [23]. The survival rate was calculated according to that mentioned in MTT assay.…”

Section: Methodsmentioning

confidence: 99%

Study on the Apoptosis Mechanism Induced by T-2 Toxin

et al. 2013

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“…The protein samples were analyzed with SDS-PAGE, after that the PVDF membrane was activated with methanol for 30 s. Then, proteins in the gel were transferred onto PVDF membrane at a current of 300 mA for 50 m, and the membrane was further blocked with 5% skim milk powder in TBST solution (8 g/L NaCl, 2.42 g/L Tris, 1 mL Tween-20, pH7.6) for 1 h. Then the membrane was soaked in 1:1000 diluted 1st antibody (the murine monoclonal antibodies against H3K4me1 to me3, H3K9me1 to me3 and H3K36me1 to me3, respectively), in TBST solution with 5% skim milk at 4 • C overnight. Following that, the membrane was eluted three times with 20 mL of TBST for 7 m each time, then the membrane was soaked into 1:5000 diluted goat anti-mouse antibody dilution at room temperature for 1 h. Finally, the membrane was washed with TBST for three times, and the color was developed with equal volume of HRP Substrate Luminol Reagent (Immobilon TM Western) and HRP Substrate Peroxide Solution (Immobilon TM Western), and photographed in GBox XT4 Chemiluminescence and Fluorescence Imaging System (Gene Company Limited, Shanghai, China) (Zhuang et al, 2011).…”

Section: Western-blotting Analysismentioning

confidence: 99%

The Methyltransferase AflSet1 Is Involved in Fungal Morphogenesis, AFB1 Biosynthesis, and Virulence of Aspergillus flavus

et al. 2020

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The filament fungal pathogen, Aspergillus flavus, spreads worldwide and contaminates several important crops. Histone posttranslational modifications are deeply involved in fungal development and virulence, but the biological function of the histone methyltransferase AflSet1 in A. flavus is still unknown. In the study, Aflset1 deletion strain was constructed through homologous recombination, and it was found that AflSet1 up-regulates hyphae growth, and promotes conidiation by sporulation regulation genes: abaA and brlA. It was also found that AflSet1 involves in sclerotia formation and AFB1 biosynthesis via sclerotia related transcriptional factors and orthodox AFB1 synthesis pathway, respectively. Crop models revealed that AflSet1 plays critical roles in colonization and AFB1 production on crop kernels. Lipase activity analysis suggested that AflSet1 affects fungal virulence to crops via digestive enzymes. Stresses tests revealed that AflSet1 is deeply involved in fungal resistance against osmotic, oxidative and cell membrane stress. The preparation of N_SET, SET domain deletion mutants and H988K mutant revealed that both domains play critical roles in fungal development and AFB1 production, and that H988 is very important in executing biological functions on morphogenesis and AFB1 synthesis. Subcellular location analysis revealed that AflSet1 is stably accumulated in nuclei in both spore germination and hyphae growth stages, even under the stress of SDS. Through immunoblot analysis, it was found that AflSet1 methylates H3K4me2 and me3 as well as H3K9me2. This study provides a solid evidence to discover the biological functions of histone methyltransferase in pathogenic fungi.

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“…Previous studies indicate that the bacterial infection can modulate The calculated data (mean ± SD) of six individuals (n = 6) with different letters (a, b, c) were significantly different (P \ 0.05) between the challenge group and the control group Characterization and expression analysis of Calmodulin (CaM) in orange-spotted grouper… the expression level of calcium vector protein (CaVP) in amphioxus (Zhuang et al 2011). In this study, we found that a significant increase of Ec-CaM transcript expression and Ec-CaM protein expression was observed in the vibrio challenge group.…”

Section: Discussionmentioning

confidence: 99%

Characterization and expression analysis of Calmodulin (CaM) in orange-spotted grouper (Epinephelus coioides) in response to Vibrio alginolyticus challenge

et al. 2015

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Vibrio alginolyticus containing the highly toxic extracellular product is one of the most serious threats to grouper survival and its minimum lethal dose is approximately 500 CFU/g fish body weight in grouper. To study the toxic effects of V. alginolyticus on the immune system in teleost, Calmodulin (CaM), an important molecular indicator gene, was cloned from the orange-spotted grouper (Epinephelus coioides). The full-length Ec-CaM consisted of a 5'-UTR of 103 bp, an ORF of 450 bp and a 3'-UTR of 104 bp. The Ec-CaM gene encoded a protein of 149 amino acids with an estimated molecular mass of 16.4 kDa and a predicted isoelectric point of 3.93. The deduced amino acid sequence showed that Ec-CaM contained four highly conserved EF-hand domains known to be critical for the function of CaM. Ec-CaM was widely expressed and the highest expression level was observed in liver. Following V. alginolyticus challenge, a sharp increase level of respiratory burst activity and apoptosis ratio were observed. Further analyses of CaM expression and p53 expression in liver, kidney and spleen by qRT-PCR demonstrated that the up-regulated expression of CaM and p53 were observed in the vibrio challenge group. Western blotting analysis confirmed that the Ec-CaM protein was strongly induced in liver at 12 h post-injection, while a sharp increase of p53 protein expression was observed at 24 h post-injection. These results showed CaM expression serving as a potential molecular indicator may help to assess the toxicological effects of V. alginolyticus on the ROS generation and apoptotic process in grouper.

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Gut CaVP is an innate immune protein against bacterial challenge in amphioxus Branchiostoma belcheri

Cited by 28 publications

References 32 publications

Study on the Apoptosis Mechanism Induced by T-2 Toxin

Study on the Apoptosis Mechanism Induced by T-2 Toxin

The Methyltransferase AflSet1 Is Involved in Fungal Morphogenesis, AFB1 Biosynthesis, and Virulence of Aspergillus flavus

Characterization and expression analysis of Calmodulin (CaM) in orange-spotted grouper (Epinephelus coioides) in response to Vibrio alginolyticus challenge

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