NELF and DSIF cause promoter proximal pausing on the hsp70 promoter in Drosophila.
Publication/Presentation Date
6-1-2003
Abstract
NELF and DSIF collaborate to inhibit elongation by RNA polymerase IIa in extracts from human cells. A multifaceted approach was taken to investigate the potential role of these factors in promoter proximal pausing on the hsp70 gene in Drosophila. Immunodepletion of DSIF from a Drosophila nuclear extract reduced the level of polymerase that paused in the promoter proximal region of hsp70. Depletion of one NELF subunit in salivary glands using RNA interference also reduced the level of paused polymerase. In vivo protein-DNA cross-linking showed that NELF and DSIF associate with the promoter region before heat shock. Immunofluorescence analysis of polytene chromosomes corroborated the cross-linking result and showed that NELF, DSIF, and RNA polymerase IIa colocalize at the hsp70 genes, small heat shock genes, and many other chromosomal locations. Finally, following heat shock induction, DSIF and polymerase but not NELF were strongly recruited to chromosomal puffs harboring the hsp70 genes. We propose that NELF and DSIF cause polymerase to pause in the promoter proximal region of hsp70. The transcriptional activator, HSF, might cause NELF to dissociate from the elongation complex. DSIF continues to associate with the elongation complex and could serve a positive role in elongation.
Volume
17
Issue
11
First Page
1402
Last Page
1414
ISSN
0890-9369
Published In/Presented At
Wu, C. H., Yamaguchi, Y., Benjamin, L. R., Horvat-Gordon, M., Washinsky, J., Enerly, E., Larsson, J., Lambertsson, A., Handa, H., & Gilmour, D. (2003). NELF and DSIF cause promoter proximal pausing on the hsp70 promoter in Drosophila. Genes & development, 17(11), 1402–1414. https://doi.org/10.1101/gad.1091403
Disciplines
Medicine and Health Sciences
PubMedID
12782658
Department(s)
Department of Medicine
Document Type
Article