Intervening sequences increase efficiency of RNA 3' processing and accumulation of cytoplasmic RNA
Open accessAbstract
Two expression vectors were constructed that differ only in the presence (+) or absence (-) of an intervening sequence (IVS) in their 5'-untranslated leaders. Transient transfection into four mammalian cell lines resulted in higher levels of the indicator protein (CAT) from the IVS(+) vector (6 to 50-fold). Cytoplasmic RNA concentrations in 293s and HeLa cell lines corresponded directly to resultant protein levels; measurements in 293s cells of transcription initiation and elongation, steady-state total nuclear RNA, and cytoplasmic RNA stability, were equivalent for the two vectors. Surprisingly, the amount of poly(A)+ nuclear RNA was greater from the IVS(+) vector. Since this difference matches the ratio seen with polyadenylated cytoplasmic RNA, our results imply that splicing is coupled to a polyadenylation/transport pathway.
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Introns possess a posttranscriptional regulatory element that efficiently induces transport of mRNA out of the nucleus and enhances mRNA stability.
“Introns are known to possess a posttranscriptional regulatory element that efficiently induces transport of mRNA out of the nucleus and enhances mRNA stability [18].”
From Optimization of AAV expression cassettes to improve packaging capacity and transgene expression in neurons (2014) · cites this paper as [18]
The Discussion (page unavailable) says IVS(+) and IVS(-) RNA had the same cytoplasmic stability and that transport rates need not have differed. Rather than attributing increased RNA accumulation to enhanced stability or faster nuclear export, the authors favor more efficient polyadenylation as the posttranscriptional mechanism. (LLM, conf=0.88)
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- Authors
- Huang, M. T. F., & Gorman, C. M.
- Affiliations
- Genentech (US)
- Corresponding author
- Gorman, Cornelia M.
- Venue
- Nucleic Acids Research
- Publisher
- Oxford University Press (OUP)
- Published
- 1990
- Volume
- 18
- Issue
- 4
- Pages
- 937-947
- ISSN
- 0305-1048 (ISSN-L)
- 1362-4962
- PMC
- PMC330348
- OpenAlex
- W2013313513
- Cited by
- 303
- References
- 0
- Citations by year
- 2024: 1 · 2022: 2 · 2021: 2 · 2020: 4 · 2019: 2 · 2018: 9 · 2017: 4 · 2016: 6 · 2015: 6 · 2014: 9 · 2013: 7 · 2012: 2
- FWCI
- 6.14
- Type
- journal article
- Language
- en
- Open access
- green
- europepmc.org
- Locations
- Nucleic Acids Research (publishedVersion)
- Europe PMC (PubMed Central) (publishedVersion)
- PubMed Central (publishedVersion)
- PubMed (publishedVersion)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA Research and Splicing
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA and protein synthesis mechanisms
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA regulation and disease
- Keywords
- introns, 5' untranslated region, RNA splicing, polyadenylation, polyadenylated RNA, RNA transport, transient transfection, nuclear RNA
- MeSH terms
- Animals, Base Sequence, Cell Line, Cell Nucleus, Cytoplasm, Genes, Bacterial, Genetic Vectors, HeLa Cells, Humans, Introns, Molecular Sequence Data, Nucleic Acid Hybridization, Plasmids, RNA, RNA Splicing, RNA, Messenger, RNA, Neoplasm, Transfection, Oligonucleotide Probes, Chloramphenicol O-Acetyltransferase
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