Position and Sequence Requirements for Poly(A) Length Regulation by the Poly(A) Limiting Element

Overview

Journal RNA

Specialty Molecular Biology

Date 2001 Jul 17

PMID 11453064

Citations 6

Authors

J D GUPTA

H Gu

D R Schoenberg

Affiliations

Soon will be listed here.

Abstract

The poly(A)-limiting element (PLE) is a cis-acting sequence that acts to limit poly(A) tail length on pre-mRNA to <20 nt. Functional PLEs are present in a number of genes, underscoring the generality of this control mechanism. The current study sought to define further the position requirements for poly(A) length regulation and the core sequence that comprises a PLE. Increasing the spacing between the PLE and the upstream 3' splice site or between the PLE and the downstream AAUAAA had no effect on poly(A) length control. However, moving the PLE from the terminal exon to either an upstream exon or intron eliminated poly(A) length control. Poly(A) length control was further evaluated using a battery of constructs in which the PLE was maintained in the terminal exon, but where upstream introns were either deleted, modified, or replaced with a polypyrimidine tract. Poly(A) length control was retained in all cases, indicating that the key feature is the presence of the PLE in the terminal exon. A battery of mutations demonstrated the importance of the 5' pyrimidine-rich portion of the element. Finally, UV crosslinking experiments identified an approximately 62-kDa protein in Hela nuclear extract that binds to a wild-type 23-nt PLE RNA oligonucleotides but not to a mutated nonfunctional form of the element.

Citing Articles

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Kini H, Kong J, Liebhaber S Mol Cell Biol. 2014; 34(7):1300-9.

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mRNA with a <20-nt poly(A) tail imparted by the poly(A)-limiting element is translated as efficiently in vivo as long poly(A) mRNA.

Peng J, Schoenberg D RNA. 2005; 11(7):1131-40.

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The poly(A)-limiting element enhances mRNA accumulation by increasing the efficiency of pre-mRNA 3' processing.

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U2AF modulates poly(A) length control by the poly(A)-limiting element.

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References

Vagner S, Vagner C, Mattaj I . The carboxyl terminus of vertebrate poly(A) polymerase interacts with U2AF 65 to couple 3'-end processing and splicing. Genes Dev. 2000; 14(4):403-13. PMC: 316384. View

Schoenberg D, Moskaitis J, SMITH Jr L, Pastori R . Extranuclear estrogen-regulated destabilization of Xenopus laevis serum albumin mRNA. Mol Endocrinol. 1989; 3(5):805-14. DOI: 10.1210/mend-3-5-805. View

Levitt N, Briggs D, Gil A, Proudfoot N . Definition of an efficient synthetic poly(A) site. Genes Dev. 1989; 3(7):1019-25. DOI: 10.1101/gad.3.7.1019. View

Pastori R, Moskaitis J, Buzek S, Schoenberg D . Coordinate estrogen-regulated instability of serum protein-coding messenger RNAs in Xenopus laevis. Mol Endocrinol. 1991; 5(4):461-8. DOI: 10.1210/mend-5-4-461. View

Schek N, Cooke C, Alwine J . Definition of the upstream efficiency element of the simian virus 40 late polyadenylation signal by using in vitro analyses. Mol Cell Biol. 1992; 12(12):5386-93. PMC: 360476. DOI: 10.1128/mcb.12.12.5386-5393.1992. View

Bauren G, Wieslander L . Splicing of Balbiani ring 1 gene pre-mRNA occurs simultaneously with transcription. Cell. 1994; 76(1):183-92. DOI: 10.1016/0092-8674(94)90182-1. View

Gunderson S, Beyer K, Martin G, Keller W, Boelens W, Mattaj L . The human U1A snRNP protein regulates polyadenylation via a direct interaction with poly(A) polymerase. Cell. 1994; 76(3):531-41. DOI: 10.1016/0092-8674(94)90116-3. View

Gilmartin G, Fleming E, Oetjen J, Graveley B . CPSF recognition of an HIV-1 mRNA 3'-processing enhancer: multiple sequence contacts involved in poly(A) site definition. Genes Dev. 1995; 9(1):72-83. DOI: 10.1101/gad.9.1.72. View

Berget S . Exon recognition in vertebrate splicing. J Biol Chem. 1995; 270(6):2411-4. DOI: 10.1074/jbc.270.6.2411. View

10.

Sittler A, Gallinaro H, Jacob M . The secondary structure of the adenovirus-2 L4 polyadenylation domain: evidence for a hairpin structure exposing the AAUAAA signal in its loop. J Mol Biol. 1995; 248(3):525-40. DOI: 10.1006/jmbi.1995.0240. View

11.

Lou H, Gagel R, Berget S . An intron enhancer recognized by splicing factors activates polyadenylation. Genes Dev. 1996; 10(2):208-19. DOI: 10.1101/gad.10.2.208. View

12.

Ruegsegger U, Beyer K, Keller W . Purification and characterization of human cleavage factor Im involved in the 3' end processing of messenger RNA precursors. J Biol Chem. 1996; 271(11):6107-13. DOI: 10.1074/jbc.271.11.6107. View

13.

Rao M, Chernokalskaya E, Schoenberg D . Regulated nuclear polyadenylation of Xenopus albumin pre-mRNA. Nucleic Acids Res. 1996; 24(20):4078-83. PMC: 146192. DOI: 10.1093/nar/24.20.4078. View

14.

Gunderson S, Vagner S, Mattaj I . Involvement of the carboxyl terminus of vertebrate poly(A) polymerase in U1A autoregulation and in the coupling of splicing and polyadenylation. Genes Dev. 1997; 11(6):761-73. DOI: 10.1101/gad.11.6.761. View

15.

Wu L, Good P, Richter J . The 36-kilodalton embryonic-type cytoplasmic polyadenylation element-binding protein in Xenopus laevis is ElrA, a member of the ELAV family of RNA-binding proteins. Mol Cell Biol. 1997; 17(11):6402-9. PMC: 232492. DOI: 10.1128/MCB.17.11.6402. View

16.

Paillard L, Omilli F, Legagneux V, Bassez T, Maniey D, Osborne H . EDEN and EDEN-BP, a cis element and an associated factor that mediate sequence-specific mRNA deadenylation in Xenopus embryos. EMBO J. 1998; 17(1):278-87. PMC: 1170378. DOI: 10.1093/emboj/17.1.278. View

17.

Antoniou M, Geraghty F, Hurst J, Grosveld F . Efficient 3'-end formation of human beta-globin mRNA in vivo requires sequences within the last intron but occurs independently of the splicing reaction. Nucleic Acids Res. 1998; 26(3):721-9. PMC: 147328. DOI: 10.1093/nar/26.3.721. View

18.

Gunderson S, Mattaj I . U1 snRNP inhibits pre-mRNA polyadenylation through a direct interaction between U1 70K and poly(A) polymerase. Mol Cell. 1998; 1(2):255-64. DOI: 10.1016/s1097-2765(00)80026-x. View

19.

Gupta J, Gu H, Chernokalskaya E, Gao X, Schoenberg D . Identification of two cis-acting elements that independently regulate the length of poly(A) on Xenopus albumin pre-mRNA. RNA. 1998; 4(7):766-76. PMC: 1369657. DOI: 10.1017/s1355838298971837. View

20.

Moreira A, Takagaki Y, Brackenridge S, Wollerton M, Manley J, Proudfoot N . The upstream sequence element of the C2 complement poly(A) signal activates mRNA 3' end formation by two distinct mechanisms. Genes Dev. 1998; 12(16):2522-34. PMC: 317083. DOI: 10.1101/gad.12.16.2522. View