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Organ
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Physiopathology
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Name

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User name	Sebastian Eilebrecht
Email	eilebrecht@ihes.fr
Phone / Fax number	+491755662397 /
Location	IHES (Systems Epigenomics Group) - 35, route de Chartres - 91440 Bures sur Yvette, France

Scientific description

7SK small-nuclear RNA has been shown to negatively regulate P-TEFb transcription elongation on the one hand and control HMGA1 transcription initiation and chromatin remodeling on the other. The non-coding 7SK RNA thereby directly interacts with both factors through different regions. While the loop 2 of the RNA specifically binds to the first HMGA1 A/T hook, thereby competing with DNA binding to the same domain, loops 1, 3 and 4 are involved in P-TEFb interaction. This raises the question of whether HMGA1 and P-TEFb cooperate during gene transcription. Using transcriptome profiling, we have identified genes that are oppositely regulated by 7SK RNA over-expression versus shRNA mediated knock-down. Furthermore, we have similarly assembled genes affected concomitantly by HMGA1 over-expression. Inhibition of P-TEFb by competitive expression of a dominant-negative Cdk9 protein leads to highly similar changes in global gene expression as the over-expression of 7SK RNA, confirming the importance of P-TEFb inhibition by 7SK RNA. HMGA1 and P-TEFb, in the case of select target genes, show strong cooperation in transcriptional activation. Finally, we provide evidence for 7SK RNA complexes containing simultaneously HMGA1 and P-TEFb. 7SK RNA thus establishes gene-dependent plasticity between HMGA1 transcription initiation and chromatin remodeling and P-TEFb transcription elongation.

Technical description

For microarray analyses, RNA amplification, labeling, hybridization and 
detection were performed following the protocols supplied by Applied 
Biosystems using the corresponding kits (Applied Biosystems, ProdNo: 
4339628 and 4336875). The data obtained were analyzed as described 
previously:
1. Eilebrecht S, BÃ©cavin C, LÃ©ger H, Benecke BJ, Benecke A (2011) HMGA1-dependent and independent 7SK RNA gene regulatory activity. RNA Biol., in the press.
2. Eilebrecht S, Brysbaert G, Wegner T, Urlaub H, Benecke BJ, Benecke A (2010) 7SK small nuclear RNA directly affects HMGA1 function in transcription regulation. Nucl. Acids Res., in the press.

References

Pubmed : http://www.ncbi.nlm.nih.gov/pubmed/21087998

Pubmed : http://www.ncbi.nlm.nih.gov/pubmed/21282977

Pubmed : http://www.ncbi.nlm.nih.gov/pubmed/21957495