Zebrafish Sp1-like protein is structurally and functionally comparable to human Sp1

Lin, Cha Jang; Hsiao, Tsun Hsien; Chung, Yi Shao; Chang, Wei-Ju; Yeh, Trai Ming; Chen, Bing Hung; Fu, Tzu Fun

doi:10.1016/j.pep.2010.10.010

Cited by 6 publications

(13 citation statements)

References 30 publications

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“…Two complementary 5=-biotinylated oligonucleotides (MDBio, Inc., Taiwain) specific to the site of interest (Table 2) were annealed and incubated in DNA affinity precipitation assay (DAPA) buffer (20 mM Tris [pH 7.4], 0.5 mM EDTA, 2 mM MgCl 2 , 60 mM KCl, 0.01% Nonidet P-40, 5% glycerol) at 4°C for 1 h with either 2 g of purified recombinant zebrafish Sp1 protein in the presence of 100 g of zebrafish liver nuclear extract or 100 g of protein in total cell lysate containing overexpressed zebrafish Sp1-like protein (15). Then, 20 l of a slurry of streptavidin-conjugated beads (Sigma-Aldrich) was added to the mixture and incubated at 4°C for another hour.…”

Section: Figmentioning

confidence: 99%

“…All three enzymes, including FDH, catalyze the production of THF. Previously, we had characterized a zebrafish Sp1-like protein and showed that this protein was structurally and functionally similar to human Sp1 (15). To examine the role of Sp1 in regulating zebrafish fdh expression, we coexpressed this Sp1-like protein in zebrafish embryos, which were injected with the fdh Ϫ1694/ ϩ58 promoter constructs simultaneously, for the promoter activity assay.…”

Section: Figmentioning

confidence: 99%

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Ethanol-Induced Upregulation of 10-Formyltetrahydrofolate Dehydrogenase Helps Relieve Ethanol-Induced Oxidative Stress

Hsiao

Lin

Chung

et al. 2014

Molecular and Cellular Biology

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Alcoholism induces folate deficiency and increases the risk for embryonic anomalies. However, the interplay between ethanol exposure and embryonic folate status remains unclear. To investigate how ethanol exposure affects embryonic folate status and one-carbon homeostasis, we incubated zebrafish embryos in ethanol and analyzed embryonic folate content and folate enzyme expression. Exposure to 2% ethanol did not change embryonic total folate content but increased the tetrahydrofolate level approximately 1.5-fold. The expression of 10-formyltetrahydrofolate dehydrogenase (FDH), a potential intracellular tetrahydrofolate reservoir, was increased in both mRNA and protein levels. Overexpressing recombinant FDH in embryos alleviated the ethanol-induced oxidative stress in ethanol-exposed embryos. Further characterization of the zebrafish fdh promoter revealed that the ؊124/؉40 promoter fragment was the minimal region required for transactivational activity. The results of site-directed mutagenesis and binding analysis revealed that Sp1 is involved in the basal level of expression of fdh but not in ethanol-induced upregulation of fdh. On the other hand, CEBP␣ was the protein that mediated the ethanol-induced upregulation of fdh, with an approximately 40-fold increase of fdh promoter activity when overexpressed in vitro. We concluded that upregulation of fdh involving CEBP␣ helps relieve embryonic oxidative stress induced by ethanol exposure.

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Section: Figmentioning

confidence: 99%

Section: Figmentioning

confidence: 99%

Ethanol-Induced Upregulation of 10-Formyltetrahydrofolate Dehydrogenase Helps Relieve Ethanol-Induced Oxidative Stress

Hsiao

Lin

Chung

et al. 2014

Molecular and Cellular Biology

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“…One of the fundamental underpinning principles is that evolution is a process in which nature needs to accomplish the duality of conserving and changing gene and protein sequences as well as structures and biological functionalities [1, 2, 9, 11, 12, 34, 47, 57]. The evolutionary dualism significantly increases the chances of having straightforward ways of dealing with complexity, change and conservation, and time.…”

Section: Resultsmentioning

confidence: 99%

“…Observations describe SP1 as regulating transcription “through synergistic effects with other transcription factors” [12, page 36] [63, 64] and supporting cofactors [47] in gene regulatory systems [2]. The role of transcription factors therefore is part of a delicate network which has to emerge entirely and simultaneously in an already existing organism in order to survive the evolutionary selection process.…”

Section: Evolutionary Issuesmentioning

confidence: 99%

“…Widespread use of zinc finger proteins in nature [2, 12], however, suggests that natural zinc finger domains have nature-given advantages and that those evolutionary traits should be replicated or reused to produce molecular tools such as zinc finger nucleases (ZFNs). Also to be considered is that single zinc fingers which contain 28–30 amino acids are simple structures with an unusual high degree of functional flexibility and structural malleability to bind distinctively any triplet depending on certain tissue condition, protein context, and form of DNA sequence.…”

Section: Introductionmentioning

confidence: 99%

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Evaluation of Novel Design Strategies for Developing Zinc Finger Nucleases Tools for Treating Human Diseases

Bach

Sherman²,

Pallis

et al. 2014

Biotechnology Research International

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Zinc finger nucleases (ZFNs) are associated with cell death and apoptosis by binding at countless undesired locations. This cytotoxicity is associated with the binding ability of engineered zinc finger domains to bind dissimilar DNA sequences with high affinity. In general, binding preferences of transcription factors are associated with significant degenerated diversity and complexity which convolutes the design and engineering of precise DNA binding domains. Evolutionary success of natural zinc finger proteins, however, evinces that nature created specific evolutionary traits and strategies, such as modularity and rank-specific recognition to cope with binding complexity that are critical for creating clinical viable tools to precisely modify the human genome. Our findings indicate preservation of general modularity and significant alteration of the rank-specific binding preferences of the three-finger binding domain of transcription factor SP1 when exchanging amino acids in the 2nd finger.

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Molecular characterization and nutritional regulation of specificity protein 1 (Sp1) in grass carp (Ctenopharyngodon idella )

Tang

et al. 2017

Aquacult Nutr

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Specificity protein 1 (Sp1) is a ubiquitous nuclear factor in eukaryotic cells and plays a crucial role in the regulation of di-/tripeptide absorption in the intestine. However, the function of Sp1 in bony fish remains poorly understood. In this study, a fish Sp1 gene (designated CiSp1) was identified from Ctenopharyngodon idella. The deduced CiSp1 protein contains structural features typical of Sp1 family proteins, including a glutamine-rich (Q-rich) region and three typical Cys2His2 zinc fingers. Quantitative real-time PCR analysis revealed ubiquitous expression of CiSp1 in all tested tissues and embryonic developmental stages of grass carp. In addition, CiSp1 expression in the intestine was significantly increased after feeding with fish meal and 270 g/kg dietary protein. Moreover, in vitro and in vivo glutamine dipeptide treatments showed similar stimulatory effects on the expression of the transcription factors CiSp1 and CiCDX2 and their potential target gene CiPepT1 in the intestines of grass carp. Finally, our results demonstrated that the mRNA expression of CiSp1/CiCDX2/CiPepT1 in the intestine was significantly increased after treatment with sodium butyrate in vitro and in vivo. Altogether, these results demonstrate for the first time that grass carp possesses a functional Sp1 and that its expression is regulated by dietary protein and feed additives. K E Y W O R D S characterization, Ctenopharyngodon idella, expression profile, nutritional regulation, specificity protein 1

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Zebrafish Sp1-like protein is structurally and functionally comparable to human Sp1

Cited by 6 publications

References 30 publications

Ethanol-Induced Upregulation of 10-Formyltetrahydrofolate Dehydrogenase Helps Relieve Ethanol-Induced Oxidative Stress

Ethanol-Induced Upregulation of 10-Formyltetrahydrofolate Dehydrogenase Helps Relieve Ethanol-Induced Oxidative Stress

Evaluation of Novel Design Strategies for Developing Zinc Finger Nucleases Tools for Treating Human Diseases

Molecular characterization and nutritional regulation of specificity protein 1 (Sp1) in grass carp (Ctenopharyngodon idella )

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