Definition of an efficient synthetic poly(A) site - PubMed (original) (raw)
Comparative Study
doi: 10.1101/gad.3.7.1019.
Affiliations
- PMID: 2570734
- DOI: 10.1101/gad.3.7.1019
Free article
Comparative Study
Definition of an efficient synthetic poly(A) site
N Levitt et al. Genes Dev. 1989 Jul.
Free article
Abstract
We constructed and analyzed a synthetic poly(A) (SPA) site that was based on the highly efficient poly(A) signal of the rabbit beta-globin gene. By use of the SPA, we demonstrate that the minimum sequences required for efficient polyadenylation are the AATAAA sequence and a GT/T-rich sequence with the correct spacing of 22-23 nucleotides between them. When placed downstream of the poly(A) site of the human alpha 2-globin gene, the SPA is used exclusively. We predict that the SPA, with its more extensive GT/T-rich sequence, is a more efficient poly(A) site than alpha-globin. Also, we compared the use of the SPA when it is placed either in the exon 3 or intron 2 of the rabbit beta-globin gene. When in the exonic position, SPA is used 10-fold more than the regular poly(A) site of rabbit beta-globin. In contrast, when it is in the intronic location, no detectable use of SPA is observed; however, the deletion of the donor site of intron 2 reactivates the intronic positioned SPA. These results indicate that the splicing of intron 2 in the rabbit beta-globin gene occurs ahead of polyadenylation and have important implications for termination of transcription. Polyadenylation, although required for termination of transcription, is not sufficient; therefore, additional termination signals for RNA polymerase II must exist.
Similar articles
- An apparent pause site in the transcription unit of the rabbit alpha-globin gene.
Vandenbergh DJ, James-Pederson M, Hardison RC. Vandenbergh DJ, et al. J Mol Biol. 1991 Jul 20;220(2):255-70. doi: 10.1016/0022-2836(91)90011-t. J Mol Biol. 1991. PMID: 1856860 - A poly(A) addition site and a downstream termination region are required for efficient cessation of transcription by RNA polymerase II in the mouse beta maj-globin gene.
Logan J, Falck-Pedersen E, Darnell JE Jr, Shenk T. Logan J, et al. Proc Natl Acad Sci U S A. 1987 Dec;84(23):8306-10. doi: 10.1073/pnas.84.23.8306. Proc Natl Acad Sci U S A. 1987. PMID: 3479794 Free PMC article. - Efficient 3'-end formation of human beta-globin mRNA in vivo requires sequences within the last intron but occurs independently of the splicing reaction.
Antoniou M, Geraghty F, Hurst J, Grosveld F. Antoniou M, et al. Nucleic Acids Res. 1998 Feb 1;26(3):721-9. doi: 10.1093/nar/26.3.721. Nucleic Acids Res. 1998. PMID: 9443963 Free PMC article. - Unexpected intron location in non-vertebrate globin genes.
Moens L, Vanfleteren J, De Baere I, Jellie AM, Tate W, Trotman CN. Moens L, et al. FEBS Lett. 1992 Nov 9;312(2-3):105-9. doi: 10.1016/0014-5793(92)80915-4. FEBS Lett. 1992. PMID: 1426238 Review. - Conserved 12-bp element downstream from mRNA polyadenylation sites.
Renan MJ. Renan MJ. Gene. 1987;60(2-3):245-54. doi: 10.1016/0378-1119(87)90233-2. Gene. 1987. PMID: 2895038 Review.
Cited by
- PUFFFIN: an ultra-bright, customisable, single-plasmid system for labelling cell neighbourhoods.
Lebek T, Malaguti M, Boezio GL, Zoupi L, Briscoe J, Elfick A, Lowell S. Lebek T, et al. EMBO J. 2024 Sep;43(18):4110-4135. doi: 10.1038/s44318-024-00154-w. Epub 2024 Jul 12. EMBO J. 2024. PMID: 38997504 Free PMC article. - Potential limitations of microdystrophin gene therapy for Duchenne muscular dystrophy.
Hart CC, Lee YI, Xie J, Gao G, Lin BL, Hammers DW, Sweeney HL. Hart CC, et al. JCI Insight. 2024 May 7;9(11):e165869. doi: 10.1172/jci.insight.165869. JCI Insight. 2024. PMID: 38713520 Free PMC article. - Harnessing eukaryotic retroelement proteins for transgene insertion into human safe-harbor loci.
Zhang X, Van Treeck B, Horton CA, McIntyre JJR, Palm SM, Shumate JL, Collins K. Zhang X, et al. Nat Biotechnol. 2024 Feb 20:10.1038/s41587-024-02137-y. doi: 10.1038/s41587-024-02137-y. Online ahead of print. Nat Biotechnol. 2024. PMID: 38379101 - Control of mammalian gene expression by modulation of polyA signal cleavage at 5' UTR.
Luo L, Jea JD, Wang Y, Chao PW, Yen L. Luo L, et al. Nat Biotechnol. 2024 Sep;42(9):1454-1466. doi: 10.1038/s41587-023-01989-0. Epub 2024 Jan 2. Nat Biotechnol. 2024. PMID: 38168982 - Regulation and function of alternative polyadenylation in development and differentiation.
Gallicchio L, Olivares GH, Berry CW, Fuller MT. Gallicchio L, et al. RNA Biol. 2023 Jan;20(1):908-925. doi: 10.1080/15476286.2023.2275109. Epub 2023 Oct 31. RNA Biol. 2023. PMID: 37906624 Free PMC article. Review.
Publication types
MeSH terms
Substances
LinkOut - more resources
Full Text Sources
Other Literature Sources
Miscellaneous