Abstract
We reported previously that authentic polyadenylation of pre-mRNAs in vitro requires at least two factors: a cleavage/specificity factor (CSF) and a fraction containing nonspecific poly(A) polymerase activity. To study the molecular mechanisms underlying 3' cleavage of pre-mRNAs, we fractionated CSF further and show that it consists of four separable subunits. One of these, called specificity factor (SF; Mr, approximately 290,000), is required for both specific cleavage and for specific polyadenylation and thus appears responsible for the specificity of the reaction. Although SF has not been purified to homogeneity, several lines of evidence suggest that it may not contain an essential RNA component. Two other factors, designated cleavage factors I (CFI; Mr, approximately 130,000) and II (CFII; Mr, approximately 110,000), are sufficient to reconstitute accurate cleavage when mixed with SF. A fourth factor, termed cleavage stimulation factor (CstF; Mr, approximately 200,000), enhances cleavage efficiency significantly when added to a mixture of the three other factors. CFI, CFII, and CstF do not contain RNA components, nor do they affect specific polyadenylation in the absence of cleavage. Although these four factors are necessary and sufficient to reconstitute efficient cleavage of one pre-RNA tested, poly(A) polymerase is also required to cleave several others. A model suggesting how these factors interact with the pre-mRNA and with each other is discussed.
References
52
Referenced
165
10.1016/0092-8674(86)90344-2
10.1016/S0092-8674(85)80007-6
10.1016/0003-2697(76)90527-3
10.1016/S0092-8674(88)91263-9
{'key': '2021111320021598000_3.11.1711.5', 'first-page': '193', 'article-title': 'Poly(A) polymerase purified from HeLa cell nuclear extract is required for both cleavage and polyadnylation of pre-mRNA in vitro.', 'volume': '9', 'year': '1989', 'journal-title': 'Mol. Cell. Biol.'}
/ Mol. Cell. Biol. / Poly(A) polymerase purified from HeLa cell nuclear extract is required for both cleavage and polyadnylation of pre-mRNA in vitro. (1989)10.1038/295572a0
10.1093/nar/11.5.1475
10.1016/S0076-6879(80)65011-3
/ Methods Enzymol. / Specific labeling of 3â² termini of RNA with T4 RNA ligase. (1980)10.1101/gad.2.5.578
10.1146/annurev.ge.20.120186.003323
10.1016/0092-8674(86)90290-4
10.1016/0168-9525(88)90028-5
10.1002/j.1460-2075.1987.tb02762.x
/ EMBO J. / Cleavage and polyadenylation of messenger RNA precursors in vitro occurs within large and specific 3â² processing complexes. (1987)10.1016/0968-0004(86)90226-4
/ Trends Biochem. Sci. / Promoter-specific activation of RNA polymerase II transcription by Sp1. (1986)10.1016/0092-8674(84)90268-X
10.1073/pnas.84.23.8408
10.1016/0092-8674(84)90551-8
10.1146/annurev.bi.55.070186.005303
10.1016/0022-2836(86)90321-9
10.1073/pnas.78.5.2737
10.1038/325673a0
10.1016/0092-8674(83)90440-3
{'key': '2021111320021598000_3.11.1711.23', 'first-page': '1', 'article-title': 'Polyadenylation of mRNA precursors.', 'volume': '950', 'year': '1988', 'journal-title': 'Biochem. Biophys. Acta'}
/ Biochem. Biophys. Acta / Polyadenylation of mRNA precursors. (1988){'key': '2021111320021598000_3.11.1711.24', 'first-page': '373', 'article-title': 'RNA sequence containing hexanucleotide AAUAAA directs efficient mRNA polyadenylation in vitro.', 'volume': '5', 'year': '1985', 'journal-title': 'Mol. Cell. Biol.'}
/ Mol. Cell. Biol. / RNA sequence containing hexanucleotide AAUAAA directs efficient mRNA polyadenylation in vitro. (1985)10.1016/S0076-6879(80)65059-9
/ Methods Enzymol. / Sequencing end-labeled DNA with base-specific chemical cleavages. (1980)10.1101/gad.2.5.588
10.1093/nar/16.12.5323
10.1093/nar/12.18.7035
10.1073/pnas.85.23.8885
10.1016/0092-8674(84)90337-4
10.1016/S0092-8674(85)80065-9
10.1002/j.1460-2075.1986.tb04446.x
/ EMBO J. / Analysis of RNA cleavage at the adenovirus-2 L3 polyadenylation site. (1986)10.1002/j.1460-2075.1988.tb03183.x
/ EMBO J. / Two proteins crosslinked to RNA containing the adenovirus L3 poly(A) site require the AAUAAA sequence for binding. (1988){'key': '2021111320021598000_3.11.1711.34', 'first-page': '226', 'article-title': 'Sedimentation analysis of polyadenylation-specific complexes.', 'volume': '8', 'year': '1988', 'journal-title': 'Mol. Cell. Biol.'}
/ Mol. Cell. Biol. / Sedimentation analysis of polyadenylation-specific complexes. (1988)10.1016/0968-0004(88)90220-4
10.1016/S0021-9258(17)39939-8
/ J. Biol. Chem. / Isolation and partial characterization of the poly(A) polymerases from HeLa cells infected with vaccinia virus. (1977)10.1146/annurev.bi.55.070186.005351
10.1038/263211a0
{'key': '2021111320021598000_3.11.1711.39', 'first-page': '1890', 'article-title': 'Mg2+ induces a sharp and reversible transition in U1 and U2 small nuclear ribonucleoprotein configurations.', 'volume': '4', 'year': '1984', 'journal-title': 'Mol. Cell. Biol.'}
/ Mol. Cell. Biol. / Mg2+ induces a sharp and reversible transition in U1 and U2 small nuclear ribonucleoprotein configurations. (1984)10.1016/0092-8674(88)90509-0
{'key': '2021111320021598000_3.11.1711.41', 'first-page': '495', 'article-title': 'Requirements for accurate and efficient mRNA 3â\x80² end cleavage and polyadenylation of a simian virus 40 early pre-RNA in vitro.', 'volume': '7', 'year': '1987', 'journal-title': 'Mol. Cell. Biol.'}
/ Mol. Cell. Biol. / Requirements for accurate and efficient mRNA 3â² end cleavage and polyadenylation of a simian virus 40 early pre-RNA in vitro. (1987){'key': '2021111320021598000_3.11.1711.42', 'first-page': '1759', 'article-title': 'Sequences downstream of AAUAAA signals affect pre-RNA cleavage and polyadenylation in vitro both directly and indirectly.', 'volume': '9', 'year': '1989', 'journal-title': 'Mol. Cell. Biol.'}
/ Mol. Cell. Biol. / Sequences downstream of AAUAAA signals affect pre-RNA cleavage and polyadenylation in vitro both directly and indirectly. (1989){'key': '2021111320021598000_3.11.1711.43', 'first-page': '4229', 'article-title': 'Multiple forms of poly(A) polymerase purified from HeLa cells function in specific mRNA 3â\x80² end formation.', 'volume': '9', 'year': '1989', 'journal-title': 'Mol. Cell. Biol.'}
/ Mol. Cell. Biol. / Multiple forms of poly(A) polymerase purified from HeLa cells function in specific mRNA 3â² end formation. (1989){'key': '2021111320021598000_3.11.1711.44', 'first-page': '1518', 'article-title': 'Products of in vitro cleavage and polyadenylation of simian virus 40 late pre-mRNAs.', 'volume': '7', 'year': '1987', 'journal-title': 'Mol. Cell. Biol.'}
/ Mol. Cell. Biol. / Products of in vitro cleavage and polyadenylation of simian virus 40 late pre-mRNAs. (1987)10.1101/gad.1.7.672
{'key': '2021111320021598000_3.11.1711.46', 'first-page': '4734', 'article-title': 'In vitro cleavage of the simian virus 40 early polyadenylation site adjacent to a required downstream TG sequence.', 'volume': '6', 'year': '1986', 'journal-title': 'Mol. Cell. Biol.'}
/ Mol. Cell. Biol. / In vitro cleavage of the simian virus 40 early polyadenylation site adjacent to a required downstream TG sequence. (1986){'key': '2021111320021598000_3.11.1711.47', 'first-page': '2052', 'article-title': 'Assembly of a polyadenylation-specific 25S ribonucleoprotein complex in vitro.', 'volume': '8', 'year': '1988', 'journal-title': 'Mol. Cell. Biol.'}
/ Mol. Cell. Biol. / Assembly of a polyadenylation-specific 25S ribonucleoprotein complex in vitro. (1988)10.1016/0092-8674(88)90411-4
10.1016/0092-8674(88)90510-7
10.1002/j.1460-2075.1987.tb02765.x
/ EMBO J. / Specific pre-cleavage and post-cleavage complexes involved in the formation of SV40 late mRNA 3â² termini in vitro. (1987){'key': '2021111320021598000_3.11.1711.51', 'first-page': '2317', 'article-title': 'The AAUAAA sequence is required both for cleavage and for polyadenylation of simian virus 40 pre-mRNA in vitro.', 'volume': '6', 'year': '1986', 'journal-title': 'Mol. Cell. Biol.'}
/ Mol. Cell. Biol. / The AAUAAA sequence is required both for cleavage and for polyadenylation of simian virus 40 pre-mRNA in vitro. (1986){'key': '2021111320021598000_3.11.1711.52', 'first-page': '3277', 'article-title': 'Identification of a complex associated with processing and polyadenylation in vitro of herpes simplex virus type 1 thymidine kinase precursor RNA.', 'volume': '7', 'year': '1987', 'journal-title': 'Mol. Cell. Biol.'}
/ Mol. Cell. Biol. / Identification of a complex associated with processing and polyadenylation in vitro of herpes simplex virus type 1 thymidine kinase precursor RNA. (1987)
Dates
Type | When |
---|---|
Created | 18 years, 2 months ago (June 5, 2007, 5:15 p.m.) |
Deposited | 3 years, 9 months ago (Nov. 13, 2021, 11:03 p.m.) |
Indexed | 1 month, 4 weeks ago (July 4, 2025, 12:03 p.m.) |
Issued | 35 years, 10 months ago (Nov. 1, 1989) |
Published | 35 years, 10 months ago (Nov. 1, 1989) |
Published Online | 35 years, 10 months ago (Nov. 1, 1989) |
Published Print | 35 years, 10 months ago (Nov. 1, 1989) |
@article{Takagaki_1989, title={Four factors are required for 3’-end cleavage of pre-mRNAs.}, volume={3}, ISSN={1549-5477}, url={http://dx.doi.org/10.1101/gad.3.11.1711}, DOI={10.1101/gad.3.11.1711}, number={11}, journal={Genes & Development}, publisher={Cold Spring Harbor Laboratory}, author={Takagaki, Y and Ryner, L C and Manley, J L}, year={1989}, month=nov, pages={1711–1724} }