A Large Fraction of Extragenic RNA Pol II Transcription Sites Overlap Enhancers
Open accessAbstract
Mammalian genomes are pervasively transcribed outside mapped protein-coding genes. One class of extragenic transcription products is represented by long non-coding RNAs (lncRNAs), some of which result from Pol_II transcription of bona-fide RNA genes. Whether all lncRNAs described insofar are products of RNA genes, however, is still unclear. Here we have characterized transcription sites located outside protein-coding genes in a highly regulated response, macrophage activation by endotoxin. Using chromatin signatures, we could unambiguously classify extragenic Pol_II binding sites as belonging to either canonical RNA genes or transcribed enhancers. Unexpectedly, 70% of extragenic Pol_II peaks were associated with genomic regions with a canonical chromatin signature of enhancers. Enhancer-associated extragenic transcription was frequently adjacent to inducible inflammatory genes, was regulated in response to endotoxin stimulation, and generated very low abundance transcripts. Moreover, transcribed enhancers were under purifying selection and contained binding sites for inflammatory transcription factors, thus suggesting their functionality. These data demonstrate that a large fraction of extragenic Pol_II transcription sites can be ascribed to cis-regulatory genomic regions. Discrimination between lncRNAs generated by canonical RNA genes and products of transcribed enhancers will provide a framework for experimental approaches to lncRNAs and help complete the annotation of mammalian genomes.
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The majority of active enhancers are themselves transcribed, producing enhancer RNAs in a bi-directional manner.
“The majority of active enhancers are themselves transcribed, producing enhancer RNAs (eRNAs) in a bi-directional manner [73–77].”
From Transcriptional enhancers and their communication with gene promoters (2021) · cites this paper as [76]
The Discussion says that non-transcribed enhancers greatly outnumber transcribed ones, contrary to the claim that a majority are transcribed. The Introduction also gives an example of unidirectional enhancer-region transcription; this paper does not establish bidirectional production as the general pattern. No page numbers are available in our copy. (LLM, conf=0.90)
Checked by a reasoning model — may be wrong.
Experimental — accuracy not yet measuredEnhancers are sites for the assembly of a complete Pol II preinitiation complex and bidirectional transcription of short-lived enhancer RNA (eRNA).
“Relatively recently, enhancers have been shown to be sites for the assembly of a complete Pol II preinitiation complex and bidirectional transcription of short-lived enhancer RNA (eRNA) [57,58,59].”
From Enhancer Function in the 3D Genome (2023) · cites this paper as [57]
The Results section on inducible upstream transcripts reports Pol II-associated enhancer transcription and very unstable noncoding RNAs; the Discussion links transcriptional directionality to control of preinitiation-complex formation. But the paper does not establish assembly of a complete preinitiation complex. Its characterized Ccl5 transcripts are strand-specific, with no signal from the opposite strand, rather than bidirectional (page numbers unavailable in our copy). (LLM, conf=0.88)
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- Authors
- De Santa, F., Barozzi, I., Mietton, F., Ghisletti, S., Polletti, S., Tusi, B. K., Muller, H., Ragoussis, J., Wei, C.-L., & Natoli, G.
- Affiliations
- IFOM (IT)
- Centre for Human Genetics (GB)
- Genome Institute of Singapore (SG)
- Corresponding author
- Natoli, Gioacchino
- ORCID
- Venue
- PLoS Biology
- Publisher
- Public Library of Science (PLoS)
- Published
- 2010-05-11
- Volume
- 8
- Issue
- 5
- Pages
- e1000384
- ISSN
- 1544-9173 (ISSN-L)
- 1545-7885
- PMC
- PMC2867938
- OpenAlex
- W1978901723
- Cited by
- 934
- References
- 80
- Citations by year
- 2026: 23 · 2025: 46 · 2024: 40 · 2023: 57 · 2022: 61 · 2021: 81 · 2020: 70 · 2019: 45 · 2018: 57 · 2017: 63 · 2016: 67 · 2015: 75 · 2014: 73 · 2013: 73 · 2012: 49
- FWCI
- 22.68
- Type
- journal article
- Language
- en
- Open access
- gold
- doi.org
- Locations
- PLoS Biology (publishedVersion, cc-by)
- PubMed Central (publishedVersion, cc-by)
- PubMed (publishedVersion)
- Archivio Istituzionale della Ricerca (Universita Degli Studi Di Milano) (submittedVersion, other-oa)
- DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
- Figshare (submittedVersion, cc-by)
- Funders
- Wellcome Trust (075491, 075491/Z/04,)
- European Commission (222008)
- Ministry of Education, Culture, Sports, Science and Technology
- Associazione Italiana per la Ricerca sul Cancro
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Genomics and Chromatin Dynamics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Cancer Research › Cancer-related molecular mechanisms research
- Life Sciences › Agricultural and Biological Sciences › Plant Science › Chromosomal and Genetic Variations
- Keywords
- RNA polymerase II, long non-coding RNA, transcribed enhancers, macrophage activation, lipopolysaccharide response, purifying selection, cis-regulatory elements, genome annotation, chromatin signatures
- MeSH terms
- Animals, Binding Sites, Female, Gene Expression Regulation, Humans, Lipopolysaccharides, Macrophage Activation, Promoter Regions, Genetic, Regulatory Sequences, Nucleic Acid, RNA Polymerase II, Transcription, Genetic, RNA, Untranslated, Mice
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