scholarly journals DNA-dependent protein kinase interacts functionally with the RNA polymerase II complex recruited at the human immunodeficiency virus (HIV) long terminal repeat and plays an important role in HIV gene expression

2011 ◽  
Vol 92 (7) ◽  
pp. 1710-1720 ◽  
Author(s):  
Shilpi Tyagi ◽  
Alex Ochem ◽  
Mudit Tyagi
Keyword(s):  
Gene Expression ◽  
Human Immunodeficiency Virus ◽  
Protein Kinase ◽  
Rna Polymerase ◽  
Rna Polymerase Ii ◽  
Dependent Protein Kinase ◽  
Immunodeficiency Virus ◽  
Target Sites ◽  
Dependent Protein ◽  
Rnap Ii

DNA-dependent protein kinase (DNA-PK), a nuclear protein kinase that specifically requires association with DNA for its kinase activity, plays important roles in the regulation of different DNA transactions, including transcription, replication and DNA repair, as well as in the maintenance of telomeres. Due to its large size, DNA-PK is also known to facilitate the activities of other factors by providing the docking platform at their site of action. In this study, by running several chromatin immunoprecipitation assays, we demonstrate the parallel distribution of DNA-PK with RNA polymerase II (RNAP II) along the human immunodeficiency virus (HIV) provirus before and after activation with tumour necrosis factor alpha. The association between DNA-PK and RNAP II is also long-lasting, at least for up to 4 h (the duration analysed in this study). Knockdown of endogenous DNA-PK using specific small hairpin RNAs expressed from lentiviral vectors resulted in significant reduction in HIV gene expression and replication, demonstrating the importance of DNA-PK for HIV gene expression. Sequence analysis of the HIV-1 Tat protein revealed three potential target sites for phosphorylation by DNA-PK and, by using kinase assays, we confirmed that Tat is an effective substrate of DNA-PK. Through peptide mapping, we found that two of these three potential phosphorylation sites are recognized and phosphorylated by DNA-PK. Mutational studies on the DNA-PK target sites of Tat further demonstrated the functional significance of the Tat–DNA-PK interaction. Thus, overall our results clearly demonstrate the functional interaction between DNA-PK and RNAP II during HIV transcription.

scholarly journals Stimulation of the DNA-dependent Protein Kinase by RNA Polymerase II Transcriptional Activator Proteins

1995 ◽  
Vol 270 (3) ◽  
pp. 1449-1454 ◽  
Author(s):  
Scott R. Peterson ◽  
Stephen A. Jesch ◽  
Thomas N. Chamberlin ◽  
Arik Dvir ◽  
Sridhar K. Rabindran ◽  
...  
Keyword(s):  
Protein Kinase ◽  
Rna Polymerase ◽  
Rna Polymerase Ii ◽  
Transcriptional Activator ◽  
Dependent Protein Kinase ◽  
Activator Proteins ◽  
Dependent Protein ◽  
Stimulation Of

scholarly journals DNA dependent protein kinase (DNA-PK) enhances HIV transcription by promoting RNA polymerase II activity and recruitment of transcription machinery at HIV LTR

Oncotarget ◽  
2020 ◽  
Vol 11 (7) ◽  
pp. 699-726 ◽  
Author(s):  
Sonia Zicari ◽  
Adhikarimayum Lakhikumar Sharma ◽  
Geetaram Sahu ◽  
Larisa Dubrovsky ◽  
Lin Sun ◽  
...  
Keyword(s):  
Protein Kinase ◽  
Rna Polymerase ◽  
Rna Polymerase Ii ◽  
Dependent Protein Kinase ◽  
Hiv Transcription ◽  
Transcription Machinery ◽  
Dependent Protein

scholarly journals GAL4 Is Regulated by the RNA Polymerase II Holoenzyme–Associated Cyclin-Dependent Protein Kinase SRB10/CDK8

1999 ◽  
Vol 3 (5) ◽  
pp. 673-678 ◽  
Author(s):  
Martin Hirst ◽  
Michael S Kobor ◽  
Neena Kuriakose ◽  
Jack Greenblatt ◽  
Ivan Sadowski
Keyword(s):  
Protein Kinase ◽  
Rna Polymerase ◽  
Rna Polymerase Ii ◽  
Dependent Protein Kinase ◽  
Dependent Protein ◽  
Rna Polymerase Ii Holoenzyme

cDNA Cloning and Gene Expression of Human Type Iα cGMP-Dependent Protein Kinase

Hypertension ◽  
1996 ◽  
Vol 27 (3) ◽  
pp. 552-557 ◽  
Author(s):  
Naohisa Tamura ◽  
Hiroshi Itoh ◽  
Yoshihiro Ogawa ◽  
Osamu Nakagawa ◽  
Masaki Harada ◽  
...  
Keyword(s):  
Gene Expression ◽  
Protein Kinase ◽  
Cdna Cloning ◽  
Dependent Protein Kinase ◽  
Human Type ◽  
Cgmp Dependent Protein Kinase ◽  
Dependent Protein

scholarly journals Chronic Elevation of Calmodulin in the Ventricles of Transgenic Mice Increases the Autonomous Activity of Calmodulin-Dependent Protein Kinase II, Which Regulates Atrial Natriuretic Factor Gene Expression

2000 ◽  
Vol 14 (8) ◽  
pp. 1125-1136 ◽  
Author(s):  
Josep M. Colomer ◽  
Anthony R. Means
Keyword(s):  
Gene Expression ◽  
Protein Kinase ◽  
Transgenic Mice ◽  
Atrial Natriuretic Factor ◽  
Ventricular Myocytes ◽  
Dependent Protein Kinase ◽  
Natriuretic Factor ◽  
Dependent Protein ◽  
Autonomous Activity

Abstract Although isoforms of Ca2+/calmodulin-dependent protein kinase II (CaMKII) have been implicated in the regulation of gene expression in cultured cells, this issue has yet to be addressed in vivo. We report that the overexpression of calmodulin in ventricular myocytes of transgenic mice results in an increase in the Ca2+/calmodulin-independent activity of endogenous CaMKII. The calmodulin transgene is regulated by a 500-bp fragment of the atrial natriuretic factor (ANF) gene promoter which, based on cell transfection studies, is itself known to be regulated by CaMKII. The increased autonomous activity of CaMKII maintains the activity of the transgene and establishes a positive feedforward loop, which also extends the temporal expression of the endogenous ANF promoter in ventricular myocytes. Both the increased activity of CaMKII and transcriptional activation of ANF are highly selective responses to the chronic overexpression of calmodulin. These results indicate that CaMKII can regulate gene expression in vivo and suggest that this enzyme may represent the Ca2+-dependent target responsible for reactivation of the ANF gene during ventricular hypertrophy.

scholarly journals The Role of RNA Polymerase II Contiguity and Long-Range Interactions in the Regulation of Gene Expression in Human Pluripotent Stem Cells

2019 ◽  
Vol 2019 ◽  
pp. 1-12
Author(s):  
Livia Eiselleova ◽  
Viktor Lukjanov ◽  
Simon Farkas ◽  
David Svoboda ◽  
Karel Stepka ◽  
...  
Keyword(s):  
Gene Expression ◽  
Stem Cells ◽  
Rna Polymerase ◽  
Rna Polymerase Ii ◽  
Long Range ◽  
Pluripotent Stem Cells ◽  
Human Pluripotent Stem Cells ◽  
Transcription Factories ◽  
Long Range Interactions ◽  
Rnap Ii

The eukaryotic nucleus is a highly complex structure that carries out multiple functions primarily needed for gene expression, and among them, transcription seems to be the most fundamental. Diverse approaches have demonstrated that transcription takes place at discrete sites known as transcription factories, wherein RNA polymerase II (RNAP II) is attached to the factory and immobilized while transcribing DNA. It has been proposed that transcription factories promote chromatin loop formation, creating long-range interactions in which relatively distant genes can be transcribed simultaneously. In this study, we examined long-range interactions between the POU5F1 gene and genes previously identified as being POU5F1 enhancer-interacting, namely, CDYL, TLE2, RARG, and MSX1 (all involved in transcriptional regulation), in human pluripotent stem cells (hPSCs) and their early differentiated counterparts. As a control gene, RUNX1 was used, which is expressed during hematopoietic differentiation and not associated with pluripotency. To reveal how these long-range interactions between POU5F1 and the selected genes change with the onset of differentiation and upon RNAP II inhibition, we performed three-dimensional fluorescence in situ hybridization (3D-FISH) followed by computational simulation analysis. Our analysis showed that the numbers of long-range interactions between specific genes decrease during differentiation, suggesting that the transcription of monitored genes is associated with pluripotency. In addition, we showed that upon inhibition of RNAP II, long-range associations do not disintegrate and remain constant. We also analyzed the distance distributions of these genes in the context of their positions in the nucleus and revealed that they tend to have similar patterns resembling normal distribution. Furthermore, we compared data created in vitro and in silico to assess the biological relevance of our results.

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