Home LiteratureArticle Details
PMID: 2170946 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Point mutations in AAUAAA and the poly (A) addition site: effects on the accuracy and efficiency of cleavage and polyadenylation in vitro.

Nucleic acids research ·Vol. 18 ·No. 19 ·1990-10-11 ·Pages 5799-805

Sheets MD, Ogg SC, Wickens MP

Abstract

Three sequences in the vicinity of poly (A) addition sites are conserved among vertebrate mRNAs. We analyze the effects of single base changes in each position of AAUAAA and in the nucleotide to which poly (A) is added on 3' end formation in vitro. All 18 possible single base changes of the AAUAAA sequence greatly reduce addition of poly (A) to RNAs that end at the poly (A) addition site, and prevent cleavage of RNAs that extend beyond. The magnitude of reduction varies greatly with the position changed and the base introduced. For any given mutation, cleavage and polyadenylation are reduced to similar extents, strongly suggesting that the same factor interacts with AAUAAA in both reactions. Mutations at and near the conserved adenosine to which poly (A) is added disturb the accuracy, but not the efficiency, of 3' end formation. For example, point mutations at the conserved adenosine shift the 3' end of the most abundant 5' half-molecule downstream by a single nucleotide. The mechanism by which these mutations might exert their effects on the precision of 3' end formation are discussed.

MeSH Terms
Base Sequence Molecular Sequence Data Mutation Poly A/genetics,metabolism RNA, Messenger/genetics,metabolism RNA, Viral/genetics,metabolism Simian virus 40/genetics
Chemicals
RNA, Messenger RNA, Viral Poly A
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Sheets M D
Cell and Molecular Biology Program, College of Agriculture and Life Sciences, University of Wisconsin, Madison 53706.
Ogg S C
Wickens M P
References (33)
33 references, click to expand
  1. 3' non-coding region sequences in eukaryotic messenger RNA.
    Nature. 1976 Sep 16;263(5574):211-4 PMID: 822353
  2. The use of thin acrylamide gels for DNA sequencing.
    FEBS Lett. 1978 Mar 1;87(1):107-10 PMID: 631324
  3. Efficient in vitro synthesis of biologically active RNA and RNA hybridization probes from plasmids containing a bacteriophage SP6 promoter.
    Nucleic Acids Res. 1984 Sep 25;12(18):7035-56 PMID: 6091052
  4. Role of the conserved AAUAAA sequence: four AAUAAA point mutants prevent messenger RNA 3' end formation.
    Science. 1984 Nov 30;226(4678):1045-51 PMID: 6208611
  5. Rapid and efficient site-specific mutagenesis without phenotypic selection.
    Proc Natl Acad Sci U S A. 1985 Jan;82(2):488-92 PMID: 3881765
  6. Separation and characterization of a poly(A) polymerase and a cleavage/specificity factor required for pre-mRNA polyadenylation.
    Cell. 1988 Mar 11;52(5):731-42 PMID: 2830992
  7. Polyadenylation of mRNA precursors.
    Biochim Biophys Acta. 1988 May 6;950(1):1-12 PMID: 2896017
  8. Multiple factors are required for specific RNA cleavage at a poly(A) addition site.
    Genes Dev. 1988 May;2(5):578-87 PMID: 2838381
  9. 3' cleavage and polyadenylation of mRNA precursors in vitro requires a poly(A) polymerase, a cleavage factor, and a snRNP.
    Cell. 1988 Sep 9;54(6):875-89 PMID: 2842067
  10. A beginning to the biochemistry of polyadenylation.
    Trends Genet. 1988 Sep;4(9):243-5 PMID: 2853468
  11. Functional analysis of point mutations in the AAUAAA motif of the SV40 late polyadenylation signal.
    Nucleic Acids Res. 1989 May 25;17(10):3899-908 PMID: 2543957
  12. Four factors are required for 3'-end cleavage of pre-mRNAs.
    Genes Dev. 1989 Nov;3(11):1711-24 PMID: 2558045
  13. Two phases in the addition of a poly(A) tail.
    Genes Dev. 1989 Sep;3(9):1401-12 PMID: 2575065
  14. An ordered pathway of assembly of components required for polyadenylation site recognition and processing.
    Genes Dev. 1989 Dec;3(12B):2180-90 PMID: 2628166
  15. Polyadenylation of mRNA: minimal substrates and a requirement for the 2' hydroxyl of the U in AAUAAA.
    Mol Cell Biol. 1990 Apr;10(4):1705-13 PMID: 1969611
  16. RNA sequence containing hexanucleotide AAUAAA directs efficient mRNA polyadenylation in vitro.
    Mol Cell Biol. 1985 Feb;5(2):373-9 PMID: 2579321
  17. Transcription termination and 3' processing: the end is in site!
    Cell. 1985 Jun;41(2):349-59 PMID: 2580642
  18. The consensus sequence YGTGTTYY located downstream from the AATAAA signal is required for efficient formation of mRNA 3' termini.
    Nucleic Acids Res. 1985 Feb 25;13(4):1347-68 PMID: 2987822
  19. Accurate cleavage and polyadenylation of exogenous RNA substrate.
    Cell. 1985 Jul;41(3):845-55 PMID: 2408761
  20. Thalassemia due to a mutation in the cleavage-polyadenylation signal of the human beta-globin gene.
    EMBO J. 1985 Feb;4(2):453-6 PMID: 4018033
  21. Generation of histone mRNA 3' ends by endonucleolytic cleavage of the pre-mRNA in a snRNP-dependent in vitro reaction.
    EMBO J. 1986 Jun;5(6):1319-26 PMID: 3015597
  22. Sequence of two related P-450 mRNAs transcriptionally increased during rat development. An R.dre.1 sequence occupies the complete 3' untranslated region of a liver mRNA.
    J Biol Chem. 1986 Aug 15;261(23):10667-72 PMID: 3015936
  23. Definition of essential sequences and functional equivalence of elements downstream of the adenovirus E2A and the early simian virus 40 polyadenylation sites.
    Mol Cell Biol. 1985 Nov;5(11):2975-83 PMID: 3018490
  24. Analysis of RNA cleavage at the adenovirus-2 L3 polyadenylation site.
    EMBO J. 1986 Aug;5(8):1929-38 PMID: 3019671
  25. The AAUAAA sequence is required both for cleavage and for polyadenylation of simian virus 40 pre-mRNA in vitro.
    Mol Cell Biol. 1986 Jul;6(7):2317-23 PMID: 3023928
  26. Formation of mRNA 3' termini: stability and dissociation of a complex involving the AAUAAA sequence.
    EMBO J. 1987 Jan;6(1):177-86 PMID: 2438129
  27. Products of in vitro cleavage and polyadenylation of simian virus 40 late pre-mRNAs.
    Mol Cell Biol. 1987 Apr;7(4):1518-29 PMID: 3037325
  28. Electrophoretic separation of polyadenylation-specific complexes.
    Genes Dev. 1987 Sep;1(7):672-82 PMID: 3428596
  29. The sequence 5'-AAUAAA-3'forms parts of the recognition site for polyadenylation of late SV40 mRNAs.
    Cell. 1981 Apr;24(1):251-60 PMID: 6113054
  30. Accurate transcription initiation by RNA polymerase II in a soluble extract from isolated mammalian nuclei.
    Nucleic Acids Res. 1983 Mar 11;11(5):1475-89 PMID: 6828386
  31. Accurate and specific polyadenylation of mRNA precursors in a soluble whole-cell lysate.
    Cell. 1983 Jun;33(2):595-605 PMID: 6134588
  32. Inhibition of RNA cleavage but not polyadenylation by a point mutation in mRNA 3' consensus sequence AAUAAA.
    Nature. 1983 Oct 13-19;305(5935):600-5 PMID: 6194440
  33. Alpha-thalassaemia caused by a polyadenylation signal mutation.
    Nature. 1983 Nov 24-30;306(5941):398-400 PMID: 6646217
Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
0305-1048
DOI
10.1093/nar/18.19.5799
Published
1990-10-11
Pages
5799-805
Language
English
Region
England
NLM ID
0411011
PMCID
PMC332317
Subset
IM
Grants
NCI NIH HHS · CA07107 · United States
NIGMS NIH HHS · GM31892 · United States
NCRR NIH HHS · S10-RR01684 · United States
External Links
PubMed source DOI: 10.1093/nar/18.19.5799 PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: [email protected]

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2