Spt6 enhances the elongation rate of RNA polymerase II in vivo
Open accessAbstract
Several eukaryotic transcription factors have been shown to modulate the elongation rate of RNA polymerase II (Pol II) on naked or chromatin-reconstituted templates in vitro. However, none of the tested factors have been shown to directly affect the elongation rate of Pol II in vivo. We performed a directed RNAi knock-down (KD) screen targeting 141 candidate transcription factors and identified multiple factors, including Spt6, that alter the induced Hsp70 transcript levels in Drosophila S2 cells. Spt6 is known to interact with both nucleosome structure and Pol II, and it has properties consistent with having a role in elongation. Here, ChIP assays of the first wave of Pol II after heat shock in S2 cells show that KD of Spt6 reduces the rate of Pol II elongation. Also, fluorescence recovery after photobleaching assays of GFP-Pol II in salivary gland cells show that this Spt6-dependent effect on elongation rate persists during steady-state-induced transcription, reducing the elongation rate from approximately 1100 to 500 bp/min. Furthermore, RNAi depletion of Spt6 reveals its broad requirement during different stages of development.
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FRAP with EGFP-tagged RNA Pol II on Hsp70 genes following heat shock in Drosophila calculated RNA Pol II speed between 1.1 and 1.5 kb/min.
“The authors calculated an RNA Pol II speed between 1.1 to 1.5 kb/min when measured on the Hsp70 genes following heat shock in Drosophila (Yao et al, 2007; Ardehali et al, 2009).”
From RNA polymerase II speed: a key player in controlling and adapting transcriptome composition (2021) · cites this paper as Ardehali et al, 2009
In the section on steady-state Hsp70 transcription (page unavailable), the paper reports FRAP of EGFP-tagged Pol II after heat-shock induction and estimates about 1.1 kb/min in control glands. It does not report a FRAP-derived speed of 1.5 kb/min, so the full stated range is not supported by this source. (LLM, conf=0.85)
Experimental — accuracy not yet measuredIn Drosophila, Spt6 increases RNA Pol II speed.
“In Drosophila, Spt6 has also been shown to increase RNA Pol II speed (Ardehali et al, 2009).”
From RNA polymerase II speed: a key player in controlling and adapting transcriptome composition (2021) · cites this paper as Ardehali et al, 2009
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References 63 · Cited by 157
- Authors
- Ardehali, M. B., Yao, J., Adelman, K., Fuda, N. J., Petesch, S. J., Webb, W. W., & Lis, J. T.
- Affiliations
- Cornell University (US)
- National Institutes of Health (US)
- Corresponding author
- Lis, John T
- Venue
- The EMBO Journal
- Published
- 2009-03-12
- Volume
- 28
- Issue
- 8
- Pages
- 1067-1077
- ISSN
- 0261-4189 (ISSN-L)
- 1460-2075
- PMC
- PMC2683705
- OpenAlex
- W2046714245
- Cited by
- 157
- References
- 63
- Citations by year
- 2025: 8 · 2024: 13 · 2023: 10 · 2022: 8 · 2021: 11 · 2020: 8 · 2019: 12 · 2018: 9 · 2017: 4 · 2016: 7 · 2015: 7 · 2014: 8 · 2013: 9 · 2012: 7
- FWCI
- 5.78
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Licence
- Locations
- The EMBO Journal (publishedVersion)
- Europe PMC (PubMed Central) (publishedVersion)
- PubMed Central (publishedVersion)
- PubMed (publishedVersion)
- Funders
- National Science Foundation (0242328)
- National Institute of General Medical Sciences (R01GM025232, R37GM025232, GM-25232)
- Division of Chemistry
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Genomics and Chromatin Dynamics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Nuclear Structure and Function
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA Research and Splicing
- Keywords
- Spt6, RNA polymerase II, transcription elongation, elongation rate, transcription factors, HSP70, heat shock response, nucleosome structure, Drosophila S2 cells, salivary glands, development (biology)
- MeSH terms
- Animals, Cell Line, Chromosomal Proteins, Non-Histone, Drosophila melanogaster, Heat-Shock Proteins, Histones, Peptide Elongation Factors, Recombinant Fusion Proteins, RNA Polymerase II, Transcription, Genetic, HSP70 Heat-Shock Proteins, Drosophila Proteins, Animals, Genetically Modified, RNA Interference, Transcriptional Elongation Factors, Fluorescence Recovery After Photobleaching
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