Abstract
Complexes form between processing factors present in a crude nuclear extract from HeLa cells and a simian virus 40 (SV40) late pre-mRNA which spans the polyadenylation [poly(A)] site. A specific 'pre-cleavage complex' forms on the pre-mRNA before cleavage. Formation of this complex requires the highly conserved sequence AAUAAA: it is prevented by mutations in AAUAAA, and by annealing DNA oligonucleotides to that sequence. After cleavage, the 5' half-molecule is found in a distinct 'post-cleavage complex'. In contrast, the 3' half-molecule is released. After cleavage and polyadenylation, polyadenylated RNA also is released. De novo formation of the post-cleavage complex requires AAUAAA and a nearby 3' terminus. Competition experiments suggest that a component which recognizes AAUAAA is required for formation of both pre- and post-cleavage complexes.
MeSH Terms
Base Sequence
Cell Nucleus/metabolism
HeLa Cells/metabolism
Humans
Plasmids
RNA Precursors/genetics
RNA Processing, Post-Transcriptional
RNA, Messenger/genetics
RNA, Viral/genetics
Simian virus 40/genetics
Chemicals
RNA Precursors
RNA, Messenger
RNA, Viral
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Zarkower D
Department of Biochemistry, College of Agriculture and Life Sciences, University of Wisconsin-Madison 53706.
Wickens M
References (17)
17 references, click to expand
-
Electrophoretic characterization of bacterial polyribosomes in agarose-acrylamide composite gels.
J Mol Biol. 1969 Apr 14;41(1):139-47
PMID: 4979520
-
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
-
Accurate and specific polyadenylation of mRNA precursors in a soluble whole-cell lysate.
Cell. 1983 Jun;33(2):595-605
PMID: 6134588
-
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
-
Recognition of cap structure in splicing in vitro of mRNA precursors.
Cell. 1984 Oct;38(3):731-6
PMID: 6567484
-
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
-
RNA sequence containing hexanucleotide AAUAAA directs efficient mRNA polyadenylation in vitro.
Mol Cell Biol. 1985 Feb;5(2):373-9
PMID: 2579321
-
Transcription termination and 3' processing: the end is in site!
Cell. 1985 Jun;41(2):349-59
PMID: 2580642
-
A sequence downstream of A-A-U-A-A-A is required for formation of simian virus 40 late mRNA 3' termini in frog oocytes.
Proc Natl Acad Sci U S A. 1985 Jun;82(12):3949-53
PMID: 2987956
-
Accurate cleavage and polyadenylation of exogenous RNA substrate.
Cell. 1985 Jul;41(3):845-55
PMID: 2408761
-
Specific small nuclear RNAs are associated with yeast spliceosomes.
Cell. 1986 Jun 20;45(6):869-77
PMID: 3518951
-
Electrophoretic separation of complexes involved in the splicing of precursors to mRNAs.
Cell. 1986 Sep 12;46(6):845-55
PMID: 2944598
-
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
-
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
-
Identification of a complex associated with processing and polyadenylation in vitro of herpes simplex virus type 1 thymidine kinase precursor RNA.
Mol Cell Biol. 1987 Sep;7(9):3277-86
PMID: 2823124
-
Electrophoretic separation of polyadenylation-specific complexes.
Genes Dev. 1987 Sep;1(7):672-82
PMID: 3428596
-
Analysis of mRNA 3' end formation by modification interference: the only modifications which prevent processing lie in AAUAAA and the poly(A) site.
EMBO J. 1987 Dec 20;6(13):4177-84
PMID: 3443104