Promoter-proximal pausing of RNA polymerase II defines a general rate-limiting step after transcription initiation.
Open Access
- 1 March 1995
- journal article
- Published by Cold Spring Harbor Laboratory in Genes & Development
- Vol. 9 (5) , 559-572
- https://doi.org/10.1101/gad.9.5.559
Abstract
We have shown previously that the majority of RNA polymerase II complexes initiated at the c-myc gene are paused in the promoter-proximal region, similar to observations in the Drosophila hsp70 gene. Our analyses define the TATA box or initiator sequences in the c-myc gene as necessary components for the establishment of paused RNA polymerase II. Deletion of upstream sequences or even the TATA box does not influence significantly the degree of transcriptional initiation or pausing. Deletion of both the TATA box and sequences at the transcription initiation site, however, abolishes transcriptional pausing of transcription complexes but still allows synthesis of full-length RNA. Further analyses with synthetic promoter constructs reveal that the simple combination of upstream activator with TATA consensus sequences or initiator sequences act synergistically to recruit high levels of RNA polymerase II complexes. Only a minor fraction of these complexes escapes into regions further downstream. Several different trans-activation domains fused to GAL4-DNA-binding domains, including strong activators such as VP16, do not eliminate promoter-proximal pausing of RNA polymerase. Thus, we conclude that pausing of RNA polymerase II is a common phenomenon in eukaryotic transcription and does not require complex promoter structures. Further analyses reveal that enhancers have a modest influence on transcription initiation and on release of transcription complexes out of the pause site but may function primarily to increase the elongation competence of transcription complexes.Keywords
This publication has 52 references indexed in Scilit:
- Phosphorylation of RNA polymerase II C-terminal domain and transcriptional elongationNature, 1994
- Transcription factor IIE binds preferentially to RNA polymerase IIa and recruits TFIIH: a model for promoter clearance.Genes & Development, 1994
- In vivo transcriptional pausing and cap formation on three Drosophila heat shock genes.Proceedings of the National Academy of Sciences, 1993
- Synergism in transcriptional activation: a kinetic view.Genes & Development, 1993
- DNA sequence requirements for generating paused polymerase at the start of hsp70.Genes & Development, 1992
- Transcription on nucleosomal templates by RNA polymerase II in vitro: inhibition of elongation with enhancement of sequence-specific pausing.Genes & Development, 1991
- Activation domains of stably bound GAL4 derivatives alleviate repression of promoters by nucleosomesCell, 1991
- The “initiator” as a transcription control elementCell, 1989
- GAL4-VP16 is an unusually potent transcriptional activatorNature, 1988
- Negative effect of the transcriptional activator GAL4Nature, 1988