Chromatin immunoprecipitation (ChIP) assay

2009 ◽  
Author(s):  
Zhongfu Ni ◽  
Danny W.-K. Ng ◽  
Jianxin Liu ◽  
Z. Jeffrey Chen
Keyword(s):  
Chromatin Immunoprecipitation ◽  
Chip Assay

Yeast Chromatin Immunoprecipitation (ChIP) Assay

2007 ◽  
Vol 2007 (1) ◽  
pp. pdb.prot4642-pdb.prot4642
Author(s):  
W. P. Tansey
Keyword(s):  
Chromatin Immunoprecipitation ◽  
Chip Assay

scholarly journals The Autoregulatory and Transactivating Functions of the Human Cytomegalovirus IE86 Protein Use Independent Mechanisms for Promoter Binding

2007 ◽  
Vol 81 (11) ◽  
pp. 5807-5818 ◽  
Author(s):  
Dustin T. Petrik ◽  
Kimberly P. Schmitt ◽  
Mark F. Stinski
Keyword(s):  
Human Cytomegalovirus ◽  
Protein Interactions ◽  
Chromatin Immunoprecipitation ◽  
Viral Gene Expression ◽  
Artificial Chromosome ◽  
Viral Gene ◽  
Chip Assay ◽  
Protein Protein Interactions ◽  
Multiple Sequence ◽  
Viral Genes

ABSTRACT The functions of the human cytomegalovirus (HCMV) IE86 protein are paradoxical, as it can both activate and repress viral gene expression through interaction with the promoter region. Although the mechanism for these functions is not clearly defined, it appears that a combination of direct DNA binding and protein-protein interactions is involved. Multiple sequence alignment of several HCMV IE86 homologs reveals that the amino acids 534LPIYE538 are conserved between all primate and nonprimate CMVs. In the context of a bacterial artificial chromosome (BAC), mutation of both P535 and Y537 to alanines (P535A/Y537A) results in a nonviable BAC. The defective HCMV BAC does not undergo DNA replication, although the P535A/Y537A mutant IE86 protein appears to be stably expressed. The P535A/Y537A mutant IE86 protein is able to negatively autoregulate transcription from the major immediate-early (MIE) promoter and was recruited to the MIE promoter in a chromatin immunoprecipitation (ChIP) assay. However, the P535A/Y537A mutant IE86 protein was unable to transactivate early viral genes and was not recruited to the early viral UL4 and UL112 promoters in a ChIP assay. From these data, we conclude that the transactivation and repressive functions of the HCMV IE86 protein can be separated and must occur through independent mechanisms.

scholarly journals PRMT5 Prevents Cardiomyocyte Hypertrophy via Symmetric Dimethylating HoxA9 and Repressing HoxA9 Expression

2020 ◽  
Vol 11 ◽  
Author(s):  
Sidong Cai ◽  
Rong Liu ◽  
Panxia Wang ◽  
Jingyan Li ◽  
Tingting Xie ◽  
...  
Keyword(s):  
Chromatin Immunoprecipitation ◽  
Adrenergic Receptor ◽  
Receptor Agonist ◽  
Therapeutic Strategies ◽  
Cardiomyocyte Hypertrophy ◽  
Chip Assay ◽  
Loss Of Function ◽  
Arginine Methyltransferase ◽  
Physical Interactions ◽  
Adrenergic Receptor Agonist

The present study reveals a link between protein arginine methyltransferase 5 (PRMT5) and Homebox A9 (HoxA9) in the regulation of cardiomyocyte hypertrophy. In cardiomyocyte hypertrophy induced by β-adrenergic receptor agonist isoprenaline (ISO), PRMT5 expression was decreased while HoxA9 was upregulated. Silencing of PRMT5 or inhibition of PRMT5 by its pharmacological inhibitor EPZ augmented the expressions of cardiomyocyte hypertrophic genes brain natriuretic peptide (BNP) and β-Myosin Heavy Chain (β-MHC), whereas overexpression of PRMT5 inhibited ISO-induced cardiomyocyte hypertrophy, suggesting that PRMT5 ameliorates cardiomyocyte hypertrophy. On the contrary, HoxA9 promoted cardiomyocyte hypertrophy, as implied by the gain-of-function and loss-of-function experiments. HoxA9 was involved in the regulation of PRMT5 in cardiomyocyte hypertrophy, since HoxA9 knockdown prevented si-RPMT5-induced cardiomyocyte hypertrophy, and HoxA9 expression impaired the anti-hypertrophic effect of PRMT5. Co-immunoprecipitation experiments revealed that there were physical interactions between PRMT5 and HoxA9. The symmetric dimethylation level of HoxA9 was decreased by ISO or EPZ treatment, suggesting that HoxA9 is methylated by PRMT5. Additionally, PRMT5 repressed the expression of HoxA9. Chromatin immunoprecipitation (ChIP) assay demonstrated that HoxA9 could bind to the promoter of BNP, and that this binding affinity was further enhanced by ISO or EPZ. In conclusion, this study suggests that PRMT5 symmetric dimethylates HoxA9 and represses HoxA9 expression, thus impairing its binding to BNP promoter and ultimately protecting against cardiomyocyte hypertrophy. These findings provide a novel insight of the mechanism underlying the cardiac protective effect of PRMT5, and suggest potential therapeutic strategies of PRMT5 activation or HoxA9 inhibition in treatment of cardiac hypertrophy.

Chromatin Immunoprecipitation (ChIP) Assay in Candida albicans

2016 ◽  
pp. 43-57 ◽  
Author(s):  
Sreyoshi Mitra ◽  
Laxmi Shanker Rai ◽  
Gautam Chatterjee ◽  
Kaustuv Sanyal
Keyword(s):  
Candida Albicans ◽  
Chromatin Immunoprecipitation ◽  
Chip Assay

scholarly journals Improvement of the Chromatin Immunoprecipitation (ChIP) Assay by DNA Fragment Size Fractionation

BioTechniques ◽  
2001 ◽  
Vol 31 (4) ◽  
pp. 740-742 ◽  
Author(s):  
Trina A. Johnson ◽  
Heather L. Wilson ◽  
William J. Roesler
Keyword(s):  
Chromatin Immunoprecipitation ◽  
Fragment Size ◽  
Chip Assay ◽  
Size Fractionation ◽  
Dna Fragment

scholarly journals MSTN mutant promoted bovine muscle satellite cell proliferation by regulating the binding of SMAD2/SMAD3 with CDKN1C

2021 ◽  
Author(s):  
Li Gao ◽  
Miao-Miao Yang ◽  
Ming-Juan Gu ◽  
Yun-Peng Liu ◽  
Cai-Hong Bu ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Cell Proliferation ◽  
Chromatin Immunoprecipitation ◽  
Muscle Development ◽  
Chip Assay ◽  
Rna Seq ◽  
Wild Type ◽  
Muscle Satellite Cell ◽  
Bovine Muscle ◽  
Expression Of Genes

Abstract Background: Myostatin (MSTN), also known as growth/differentiation factor 8, mostly expressed in skeletal muscle and plays negative roles in regulation of muscle development. Previous studies had proved that MSTN have important effect on cell proliferation. Therefore we aimed to investigate the mechanism of MSTN in regulating the proliferation of bovine muscle satellite cells (MSCs).Methods: Bovine MSCs of MSTN mutant (MT) and wild type (WT) were obtained, we detected the cell proliferation and cell cycle by EdU proliferation assay and Flow cytometry. Then we detected the expression of genes associated with cell cycle by Real-time PCR and Western blotting . RNA-seq and Chromatin immunoprecipitation (ChIP)assay were performed to research the mechanism of MSTN in regulating the cell proliferation. Results: In this study, we found that MSTN mutant promoted the proliferation of MSCs. The expression of CyclinA, CyclinD and CyclinE were all increased after MSTN mutant, while the expression of CDKN1C (P57), CDKN2A, CDKN2C and CDKN2D were down-regulated, which were consistent with the promotion of cell proliferation. Among these genes, CDKN1C(P57) down-regulated most significantly. RNA-seq results showed that MSTN mutant affected the SMAD binding, so we performed ChIP-qPCR and demonstrated that the SMAD2/SMAD3 transcription factor combined with the promoter of CDKN1C thus to increase the expression of CDKN1C, this demonstrating that MSTN regulated the expression of CDKN1C through SMAD2/SMAD3 complex. Finally, overexpression of SMAD3 in wild type cells increased the expression of CDKN1C, further suggested that SMAD3 regulated the expression of CDKN1C. Conclusion: MSTN mutant down-regulated the expression of SMAD2/SMAD3, then reduced the promotion of SMAD2/SMAD3 to the expression of CDKN1C, thus to inhibit the expression of CDKN1C, then promoting the cell cycle.

HNF-1α plays an important role in IL-6-induced expression of the human angiotensinogen gene

2007 ◽  
Vol 293 (1) ◽  
pp. C401-C410 ◽  
Author(s):  
Sudhir Jain ◽  
Yanna Li ◽  
Sai Patil ◽  
Ashok Kumar
Keyword(s):  
Chromatin Immunoprecipitation ◽  
Binding Sites ◽  
Promoter Activity ◽  
Acute Phase Protein ◽  
Gene Promoter ◽  
Proximal Promoter ◽  
Chip Assay ◽  
Its Gene ◽  
Hep3b Cells ◽  
Responsive Element

Angiotensinogen (AGT) is the precursor of one of the most important vasoactive hormone angiotensin II and this gene locus is associated with human essential hypertension. AGT is an acute phase protein and its gene expression is regulated by IL-6. Previous studies have identified three potential STAT-3 binding sites (APREs) located between −160 and −280 of the hAGT gene promoter but only APRE-1 (located between −271 and −279) was shown to be a bonafide enhancer for IL-6-induced promoter activity. We show here that APRE-2, located between −236 and −247, is indeed an HNF-1α-binding site and plays an important role in basal and IL-6 induced promoter activity of this gene. Our chromatin immunoprecipitation (ChIP) assay shows that HNF-1α binds to this region of the hAGT gene promoter and its recruitment is increased in the presence of IL-6 in Hep3B cells. We also show that the promoter activity of a deletion construct containing only 223 bp of the hAGT gene promoter (that contains only APRE-3) is increased after IL-6 treatment. Our ChIP assay shows that IL-6 treatment recruits STAT-3 to APRE-3 and suggests that this is also an IL6 responsive element. We have previously shown that GR binds to the proximal promoter of the hAGT gene. Since GR physically interacts with STAT-3, we propose that transcription factors GR, STAT-3, and HNF-1α that bind to the nucleotide sequence located between −160 and −280 of the hAGT gene promoter are responsible for IL-6 induced promoter activity of this gene.

Sign in / Sign up

Export Citation Format

Share Document