AP2/ERF transcription factor is a kind of transcription factors widely existing in plants, and contains at least a conserved AP2/ERF domain composed of about 60-70 amino acids. AP2/ERF transcription factors are widely involved in a variety of physiological processes in plants, including plant development, fruit ripening, flower development and other plant development processes, as well as such stress response processes as damage, pathogen defense, high-salt condition and drought. In recent years, secondary metabolic engineering that takes transcription factors as genetic manipulation targets has developed rapidly in improving the content of active ingredients and the quality of medicinal plants. This paper reviews the recent progress in the regulation of secondary metabolites biosynthesis with AP2/ERF transcription factors, and provides theoretical basis for the exploration of efficient regulatory targets, the regulation of secondary metabolites in medicinal plants, the targeted improvement of the content of active ingredients in traditional Chinese medicine, and the sustainable supply of high-quality traditional Chinese medicines.
Gene Expression Regulation, Plant ; Phylogeny ; Plant Proteins/metabolism* ; Transcription Factor AP-2/metabolism* ; Transcription Factors/metabolism*

Activating protein-2 (AP-2) is a cell type-specific DNA binding transcription factor family with the ability to regulate the expression of specific target genes. Five isoforms of AP-2 have been discovered, they are AP-2alpha, AP-2beta, AP-2gamma, AP-2delta and AP-2epsilon. AP-2s are involved in the regulation of cell proliferation, differentiation and apoptosis as well as embryogenesis of mammary animals. Recently, the function of AP-2 in neoplasm has attracted increasing attention. Researches reveal that the modulation of AP-2 in tumorigenesis may be dual, either inhibitory or promoting, which depends on the specific tissues,stages of cancer progression and difference between five family members. This review summarizes recent research progress on the role of AP-2 in the oncogenesis and their potential applications in clinical practice.
Animals ; Cell Transformation, Neoplastic ; genetics ; Gene Expression Regulation, Neoplastic ; Humans ; Neoplasms ; etiology ; genetics ; Transcription Factor AP-2 ; genetics

Since previous studies have reported that hydroquinone (HQ) exerted immunosuppressive and anti-inflammatory activity, various HQ derivatives have been synthesized and their biological activities investigated. In this study, we explored the anti-inflammatory activity of JS-III-49, a novel HQ derivative, in macrophage-mediated inflammatory responses. JS-III-49 suppressed the production of the inflammatory mediators nitric oxide (NO) and prostaglandin E2 (PGE2) and down-regulated the mRNA expression of the inflammatory enzymes cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS) as well as the expression of the pro-inflammatory cytokines interleukin-6 (IL-6) and IL-1b without cytotoxicity in LPS-stimulated RAW264.7 cells. JS-III-49 inhibited nuclear translocation of the NF-kB transcription factors p65 and p50 by directly targeting Akt, an upstream kinase of the NF-kB pathway, in LPS-stimulated RAW264.7 cells. However, JS-III-49 did not directly inhibit the kinase activities of Src and Syk, which are upstream kinases of Akt, in LPS-stimulated RAW264.7 cells. Moreover, JS-III-49 suppressed the nuclear translocation of c-Fos, one of the components of AP-1, by specifically targeting p38, an upstream mitogen-activated protein kinase (MAPK) in the AP-1 pathway in LPS-stimulated RAW264.7 cells. These results suggest that JS-III-49 plays an anti-inflammatory role in LPS-stimulated macrophages by targeting Akt and p38 in the NF-kB and AP-1 pathways, respectively.
Cyclooxygenase 2 ; Cytokines ; Dinoprostone ; Interleukin-6 ; Macrophages ; NF-kappa B ; Nitric Oxide ; Nitric Oxide Synthase Type II ; Phosphotransferases ; Protein Kinases ; RNA, Messenger ; Transcription Factor AP-1 ; Transcription Factors

CTHRC1 (collagen triple-helix repeat-containing 1), a protein secreted during the tissue-repair process, is highly expressed in several malignant tumors, including pancreatic cancer. We recently showed that CTHRC1 has an important role in the progression and metastasis of pancreatic cancer. Although CTHRC1 secretion affects tumor cells, how it promotes tumorigenesis in the context of the microenvironment is largely unknown. Here we identified a novel role of CTHRC1 as a potent endothelial activator that promotes angiogenesis by recruiting bone marrow-derived cells to the tumor microenvironment during tumorigenesis. Recombinant CTHRC1 (rCTHRC1) enhanced endothelial cell (EC) proliferation, migration and capillary-like tube formation, which was consistent with the observed increases in neovascularization in vivo. Moreover, rCTHRC1 upregulated angiopoietin-2 (Ang-2), a Tie2 receptor ligand, through ERK-dependent activation of AP-1 in ECs, resulting in recruitment of Tie2-expressing monocytes (TEMs) to CTHRC1-overexpressing tumor tissues. Treatment with a CTHRC1-neutralizing antibody-abrogated Ang-2 expression in the ECs in vitro. Moreover, administration of a CTHRC1-neutralizing antibody to a xenograft mouse model reduced the tumor burden and infiltration of TEMs in the tumor tissues, indicating that blocking the CTHRC1/Ang-2/TEM axis during angiogenesis inhibits tumorigenesis. Collectively, our findings support the hypothesis that CTHRC1 induction of the Ang-2/Tie2 axis mediates the recruitment of TEMs, which are important for tumorigenesis and can be targeted to achieve effective antitumor responses in pancreatic cancers.
Angiopoietin-2 ; Animals ; Carcinogenesis ; Endothelial Cells ; Heterografts ; In Vitro Techniques ; Mice ; Monocytes* ; Neoplasm Metastasis ; Pancreatic Neoplasms ; Receptor, TIE-2 ; Transcription Factor AP-1 ; Tumor Burden ; Tumor Microenvironment

Several putative transcription factor binding sites (TFBSs) exist in the PCV2 rep gene promoter. To explore if porcine circovirus type 2 (PCV2) could regulate the viral replication by using these TFBSs, we conducted electrophoretic mobility shift assay (EMSA), DNA-pull down and liquid chromatography-tandem mass spectrometric (LC-MS/MS) assays. EMSA confirmed the binding activity of the rep gene promoter with nuclear proteins of host cells. DNA-pull down and LC-MS/MS identified the porcine transcription factor AP-2δ (poTFAP2δ) could bind the PCV2 rep gene promoter. Dual-luciferase reporter assay, quantitative real-time PCR, Western blotting and indirect immunofluorescent assay demonstrated that poTFAP2δ could not only promote the activity of the rep gene promoter, but also enhance the transcription/translation activity of the rep/cap gene and the virus titer of PCV2 during the entire life cycle of PCV2 infection. This study revealed the molecular mechanism of PCV2 using host proteins to enhance the viral replication, provided a new perspective for studying the pathogenic mechanism of PCV2 from virus and host interactions, and provided a theoretical basis for developing highly effective PCV2 vaccines.
Animals ; Cell Line ; Chromatography, Liquid ; Circoviridae Infections ; Circovirus ; DNA Helicases ; Diabetes Mellitus, Type 2 ; Promoter Regions, Genetic ; Swine ; Tandem Mass Spectrometry ; Transcription Factor AP-2 ; Virus Replication

PURPOSE: This study was designed to pursue the basic relationship between MMP-2 and ligament injury via ex vivo stretching of the anterior cruciate ligament (ACL) and its changes by signal transduction pathway. MATERIALS AND METHODS: After femur-ACL-tibia complex was harvested from rats, release of MMP-2 in stretch group and non-stretch group was checked using gelatin zymography. Firstly, authors investigated whether MMP-2 was released from the ligament or not and if so, how long it was released. In addition, the complexes were divided into two groups. In group I, 5 and 12N weights were used to stretch the complex for 10, 30, and 60-minute. In group II, after the ligament complexes were stretched by 15N for 30 minutes, various signal pathway inhibitors such as pertusis toxin, SP600125, PD98059, KT5720, curcumin, and Bay11-7082 were added to their supernatants. MMP-2 activity was evaluated. RESULTS: MMP-2 was immediately released after ligament injury and its activity was increased in proportion to stretching duration and magnitude. As for the signal pathways, inhibition of JNK, ERK, AP-1, and NF-k beta pathways caused MMP-2 expression to be decreased. CONCLUSION: It is considered that the release of MMP-2 plays an important role in remodeling process after ACL injury.
Animals ; Anterior Cruciate Ligament ; Anthracenes ; Carbazoles ; Curcumin ; Flavonoids ; Gelatin ; Knee ; Ligaments ; Matrix Metalloproteinase 2 ; Nitriles ; Pyrroles ; Rats ; Signal Transduction ; Sulfones ; Transcription Factor AP-1 ; Weights and Measures

Charged multivesicular body protein 5 (CHMP5) has a key role in multivesicular body biogenesis and a critical role in the downregulation of signaling pathways through receptor degradation. However, the role of CHMP5 in T-cell receptor (TCR)-mediated signaling has not been previously investigated. In this study, we utilized a short hairpin RNA-based RNA interference approach to investigate the functional role of CHMP5. Upon TCR stimulation, CHMP5-knockdown (CHMP5(KD)) Jurkat T cells exhibited activation of TCR downstream signaling molecules, such as PKCθ and IKKαβ, and resulted in the activation of nuclear factor-κB and the marked upregulation of TCR-induced gene expression. Moreover, we found that activator protein-1 and nuclear factor of activated T-cells transcriptional factors were markedly activated in CHMP5(KD) Jurkat cells in response to TCR stimulation, which led to a significant increase in interleukin-2 secretion. Biochemical studies revealed that CHMP5 endogenously forms high-molecular-weight complexes, including TCR molecules, and specifically interacts with TCRβ. Interestingly, flow cytometry analysis also revealed that CHMP5(KD) Jurkat T cells exhibit upregulation of TCR expression on the cell surface compared with control Jurkat T cells. Taken together, these findings demonstrated that CHMP5 might be involved in the homeostatic regulation of TCR on the cell surface, presumably through TCR recycling or degradation. Thus CHMP5 is implicated in TCR-mediated signaling.
Down-Regulation ; Flow Cytometry ; Gene Expression ; Humans ; Interleukin-2 ; Jurkat Cells ; Multivesicular Bodies ; Receptors, Antigen, T-Cell* ; Recycling ; RNA Interference ; T-Lymphocytes* ; Transcription Factor AP-1 ; Up-Regulation

OBJECTIVETo identify novel genetic mutations in Chinese patients with congenital patent ductus arteriosus (PDA).

METHODClinical data and peripheral blood specimens from a kindred spanning 3 generations in which 5 of 16 individuals had PDA and a cohort of 95 unrelated subjects with PDA were collected, and 100 unrelated healthy individuals were included as controls. The coding exons and flanking introns of TFAP-2B gene were amplified by polymerase chain reaction (PCR) with specific primers. We aligned the acquired sequences with which publicized in GenBank by the aid of program BLAST. Reverse transcription-polymerase chain reaction (RT-PCR) was used to amplify the parts of TFAP-2B and sequencing was performed on PCR products forward and reversely directly.

RESULTSequencing of TFAP-2B identified that there was a splice-junction in intron 3 [intron 3(+5)G > A] and a 60 bp deletion was found in exon 3 by nested PCR. Additionally, a novel single nucleotide polymorphism (SNP) where a transition of guanine (G) to adenine (A) was identified at 34 bp front of transcription initiation site in TFAP-2B gene. There were significant differences in the prevalence of alleles G and A between controls and PDA patients (Z = -2.513, P = 0.012).

CONCLUSIONWe identified a novel splice-junction in TFAP-2B gene which might lead to hereditary PDA in a Chinese family. However, the mechanism by which this mutation results in PDA is still to be ascertained.

Case-Control Studies ; Child ; Child, Preschool ; Ductus Arteriosus, Patent ; genetics ; Exons ; Female ; Humans ; Infant ; Male ; Mutation ; Transcription Factor AP-2 ; genetics

OBJECTIVETo evaluate the effect of low salt (LS) on the expression of cyclooxygenase-2 (COX-2) and the activity of nuclear factor kappa B (NF-kappaB) and activator protein-1 (AP-1) in the mouse macula densa derived (MMDD1) cell line.

METHODSMMDD1 cells were transfected with luciferase reporter plasmid containing AP-1 or NF-kappaB. Luciferase reporter assay was used to evaluate the effect of normal salt (NS) and low salt (LS) on the activities of NF-kappaB and AP-1. The changes of COX-2 expression were examined by RT-PCR. The expression of p-p38 MAPK, p-p44/42, c-Jun, c-Fos, and COX-2 in MMDD1 cells were analyzed by Western blot.

RESULTSThe expressions of COX-2 mRNA and protein in MMDD1 cells were significantly increased by LS (P < 0.01). Phosphorylated p38 and p44/42 MAP kinase were significantly increased by treatment at 180 min (P < 0.01). The up-regulated COX-2 protein expression with LS were significantly reduced with SB 203580 (p38 inhibitors) and PD-98059 (p44/42 inhibitors) (P < 0.01). The expressions of c-Jun and c-Fos were increased by LS. The luciferase activities of AP-1 and NF-kappaB were stimulated in LS (P < 0.01), the up-regulated luciferase activities were attenuated by PDTC at 25 micromol/L (NF-kappaB inhibitor) and curcumin at 20 micromol/L (AP-1 inhibitor) (P < 0.01). LS altered COX-2 mRNA abundance and protein expression were decreased in treatment with PDTC at 25 micromol/L, curcumin at 20 micromol/L (P < 0.01).

CONCLUSIONLS can induce the expression of COX-2 in MMDD1 cells, which may be involved in the activation of p38 MAP kinase, p44/42 kinase, AP-1, and NF-kappaB pathways.

Animals ; Cell Line ; Cyclooxygenase 2 ; metabolism ; Kidney ; cytology ; metabolism ; Mice ; NF-kappa B ; metabolism ; Signal Transduction ; Transcription Factor AP-1 ; metabolism

Luteolin is a flavonoid found in abundance in celery, green pepper, and dandelions. Previous studies have shown that luteolin is an anti-inflammatory and anti-oxidative agent. In this study, the anti-inflammatory capacity of luteolin and one of its glycosidic forms, luteolin-7-O-glucoside, were compared and their molecular mechanisms of action were analyzed. In lipopolysaccharide (LPS)-activated RAW 264.7 cells, luteolin more potently inhibited the production of nitric oxide (NO) and prostaglandin E2 as well as the expression of their corresponding enzymes (inducible NO synthase (iNOS) and cyclooxygenase-2 (COX-2) than luteolin-7-O-glucoside. The molecular mechanisms underlying these effects were investigated to determine whether the inflammatory response was related to the transcription factors, nuclear factor (NF)-kappaB and activator protein (AP)-1, or their upstream signaling molecules, mitogen-activated protein kinases (MAPKs) and phosphoinositide 3-kinase (PI3K). Luteolin attenuated the activation of both transcription factors, NF-kappaB and AP-1, while luteolin-7-O-glucoside only impeded NF-kappaB activation. However, both flavonoids inhibited Akt phosphorylation in a dose-dependent manner. Consequently, luteolin more potently ameliorated LPS-induced inflammation than luteolin-7-O-glucoside, which might be attributed to the differentially activated NF-kappaB/AP-1/PI3K-Akt pathway in RAW 264.7 cells.
Apium graveolens ; Capsicum ; Cyclooxygenase 2 ; Dinoprostone ; Flavones ; Flavonoids ; Glucosides ; Inflammation ; Luteolin* ; Mitogen-Activated Protein Kinases ; NF-kappa B ; Nitric Oxide ; Nitric Oxide Synthase ; Phosphorylation ; Taraxacum ; Transcription Factor AP-1 ; Transcription Factors