Complex effects of flavopiridol on the expression of primary response genes

Cell Division

Xavier Graña¹  Daphne Georlette²  Judit Garriga³  Havva Keskin³ 

[1] Department of Biochemistry, Temple University School of Medicine, Philadelphia, PA 19140, USA;Department of Biology, Temple University, 436 Biology Life Sciences Building, Philadelphia, PA 19122, USA;Fels Institute for Cancer Research and Molecular Biology, Temple University School of Medicine, AHP bldg., room 308, 3307 North Broad St, Philadelphia, PA 19140, USA

关键词: Control of gene expression;    CDKs;    RNA polymerase II;    CDK9;    Transcription;    Quiescence;    Mitogenic stimuli;    Primary Response genes;   

Others  :  792565
DOI  :  10.1186/1747-1028-7-11

 received in 2011-12-05, accepted in 2012-03-29,  发布年份 2012

【 摘 要 】

Background

The Positive Transcription Elongation Factor b (P-TEFb) is a complex of Cyclin Dependent Kinase 9 (CDK9) with either cyclins T1, T2 or K. The complex phosphorylates the C-Terminal Domain of RNA polymerase II (RNAPII) and negative elongation factors, stimulating productive elongation by RNAPII, which is paused after initiation. P-TEFb is recruited downstream of the promoters of many genes, including primary response genes, upon certain stimuli. Flavopiridol (FVP) is a potent pharmacological inhibitor of CDK9 and has been used extensively in cells as a means to inhibit CDK9 activity. Inhibition of P-TEFb complexes has potential therapeutic applications.

Results

It has been shown that Lipopolysaccharide (LPS) stimulates the recruitment of P-TEFb to Primary Response Genes (PRGs) and proposed that P-TEFb activity is required for their expression, as the CDK9 inhibitor DRB prevents localization of RNAPII in the body of these genes. We have previously determined the effects of FVP in global gene expression in a variety of cells and surprisingly observed that FVP results in potent upregulation of a number of PRGs in treatments lasting 4-24 h. Because inhibition of CDK9 activity is being evaluated in pre-clinical and clinical studies for the treatment of several pathologies, it is important to fully understand the short and long term effects of its inhibition. To this end, we determined the immediate and long-term effect of FVP in the expression of several PRGs. In exponentially growing normal human fibroblasts, the expression of several PRGs including FOS, JUNB, EGR1 and GADD45B, was rapidly and potently downregulated before they were upregulated following FVP treatment. In serum starved cells re-stimulated with serum, FVP also inhibited the expression of these genes, but subsequently, JUNB, GADD45B and EGR1 were upregulated in the presence of FVP. Chromatin Immunoprecipitation of RNAPII revealed that EGR1 and GADD45B are transcribed at the FVP-treatment time points where their corresponding mRNAs accumulate. These results suggest a possible stress response triggered by CDK9 inhibition than ensues transcription of certain PRGs.

Conclusions

We have shown that certain PRGs are transcribed in the presence of FVP in a manner that might be independent of CDK9, suggesting a possible alternative mechanism for their transcription when P-TEFb kinase activity is pharmacologically inhibited. These results also show that the sensitivity to FVP is quite variable, even among PRGs.

【 授权许可】

   
2012 Keskin et al; licensee BioMed Central Ltd.

【 图 表 】

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