Unexpected point mutations activate cryptic 3' splice sites by perturbing a natural secondary structure within a yeast intron.
10.1101/gad.5.7.1252
Crossref journal-article
Cold Spring Harbor Laboratory
Genes & Development (246)
Abstract

The 3' splice site of the budding yeast Kluyveromyces lactis actin gene (ACT) intron is distally spaced (122 nucleotides) from its branchpoint and is also preceded by a silent PyAG located 43 nucleotides upstream. We devised a genetic screen that resulted in the isolation of several randomly induced cis-acting mutations that activate the silent PyAG as a 3' splice site. These mutations fall within a region surrounding this PyAG, which can hypothetically fold into a higher-order structure. Site-directed mutational analyses demonstrate that a hairpin structure in this region is required for correct 3' splice-site selection. Analysis of the point mutations suggests that local breathing of the hairpin near the first PyAG can lead to its activation. These data demonstrate that 3' splice-site selection is not a consequence of a linear, directional scanning mechanism, but support the notion of a critical positioning requirement for 3' splice-site selection. We speculate on the possible origin of this intron-encoded structural motif, which has homology to a bacterial transposon and suggests one possible origin for alternative splicing mechanisms in higher eukaryotes.

Bibliography

Deshler, J. O., & Rossi, J. J. (1991). Unexpected point mutations activate cryptic 3’ splice sites by perturbing a natural secondary structure within a yeast intron. Genes & Development, 5(7), 1252–1263.

Authors 2
  1. J O Deshler (first)
  2. J J Rossi (additional)
References 39 Referenced 55
  1. Ausubel, F.A., R. Brent, R.E. Kingston, D.D. Moore, J.G. Seidman, J.A. Smith, and K. Struhl. 1990. Current protocols in molecular biology. Greene Publishing/Wiley-Interscience, New York.
  2. 10.1016/0092-8674(77)90272-0
  3. 10.1128/jb.143.2.971-980.1980 / J. Bacteriol. / In vitro gene fusions that join an enzymatically active β-galactosidase segment to amino-terminal fragments of exogenous proteins: Escherichia coli plasmid vectors for the detection and cloning of translational signals. (1980)
  4. 10.1002/j.1460-2075.1986.tb04317.x / EMBO J. / Yeast pre-messenger RNA splicing efficiency depends on critical spacing requirements between the branch point and 3′ splice site. (1986)
  5. {'key': '2021111320183746000_5.7.1252.5', 'first-page': '4852', 'article-title': 'The 216-nucleotide intron of the E1A pre-mRNA contains a hairpin structure that permits utilization of unusually distant branch acceptors.', 'volume': '9', 'year': '1989', 'journal-title': 'Mol. Cell. Biol.'} / Mol. Cell. Biol. / The 216-nucleotide intron of the E1A pre-mRNA contains a hairpin structure that permits utilization of unusually distant branch acceptors. (1989)
  6. {'key': '2021111320183746000_5.7.1252.6', 'first-page': '2208', 'article-title': 'Kluyveromyces lactis maintains Saccharomyces cerevisiae intron-encoded splicing signals.', 'volume': '9', 'year': '1989', 'journal-title': 'Mol. Cell. Biol.'} / Mol. Cell. Biol. / Kluyveromyces lactis maintains Saccharomyces cerevisiae intron-encoded splicing signals. (1989)
  7. 10.1016/0092-8674(84)90468-9
  8. 10.1016/0092-8674(88)90202-4
  9. {'key': '2021111320183746000_5.7.1252.9', 'first-page': '225', 'article-title': 'Effects on mRNA splicing of mutations in the 3â\x80² region of the Saccharomyces cerevisiae actin intron.', 'volume': '7', 'year': '1987', 'journal-title': 'Mol. Cell. Biol.'} / Mol. Cell. Biol. / Effects on mRNA splicing of mutations in the 3′ region of the Saccharomyces cerevisiae actin intron. (1987)
  10. 10.1016/0003-2697(86)90403-3
  11. 10.1038/340205a0
  12. 10.1101/gad.4.1.98
  13. 10.1128/jb.153.1.163-168.1983 / J. Bacteriol. / Transformation of intact yeast cells treated with alkali cations. (1983)
  14. 10.1016/0092-8674(83)90433-6
  15. {'key': '2021111320183746000_5.7.1252.15', 'first-page': '5036', 'article-title': 'Exon as well as intron sequences are cis-regulating elements for the mutually exclusive alternative splicing of the β tropomyosin gene.', 'volume': '10', 'year': '1990', 'journal-title': 'Mol. Cell. Biol.'} / Mol. Cell. Biol. / Exon as well as intron sequences are cis-regulating elements for the mutually exclusive alternative splicing of the β tropomyosin gene. (1990)
  16. Maniatis, T., E.F. Fritsch, and J. Sambrook. 1982. Molecular cloning: A laboratory manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York.
  17. Miller, J.H. 1972. Experiments in molecular genetics. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York.
  18. 10.1101/gad.4.7.1172
  19. 10.1002/j.1460-2075.1987.tb02720.x / EMBO J. / Specific accessory sequences in Saccharomyces cerevisiae introns control assembly of pre-mRNAs into spliceosomes. (1987)
  20. 10.1073/pnas.77.7.3912
  21. 10.1126/science.6206566
  22. 10.1016/0092-8674(87)90564-2
  23. 10.1016/0092-8674(91)90219-O
  24. 10.1016/0092-8674(85)90066-2
  25. 10.1016/0092-8674(86)90343-0
  26. 10.1016/0092-8674(84)90553-1
  27. 10.1038/317735a0
  28. 10.1128/jb.169.5.1836-1846.1987 / J. Bacteriol. / Nucleotide sequence analysis of RepFIC, a basic replicon present in IncFI plasmids P307 and F, and its relation to the repA replicon of IncFII plasmids. (1987)
  29. 10.1126/science.2448875
  30. {'key': '2021111320183746000_5.7.1252.30', 'first-page': '3711', 'article-title': 'Two functional α-tubulin genes of the yeast Saccharomyces cerevisiae encode divergent proteins.', 'volume': '6', 'year': '1986', 'journal-title': 'Mol. Cell. Biol.'} / Mol. Cell. Biol. / Two functional α-tubulin genes of the yeast Saccharomyces cerevisiae encode divergent proteins. (1986)
  31. 10.1002/j.1460-2075.1988.tb03101.x / EMBO J. / A U1 snRNA : Pre-mRNA base pairing interaction is required early in yeast spliceosome assembly but does not uniquely define the 5′ cleavage site. (1988)
  32. Sherman, F., G.R. Fink, and J.B. Hicks. 1986. Methods in yeast genetics. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York.
  33. 10.1101/gad.2.10.1258
  34. 10.1038/342243a0
  35. 10.1016/0092-8674(85)90239-9
  36. 10.1002/j.1460-2075.1986.tb04412.x / EMBO J. / Mutations in conserved intron sequences affect multiple steps in the yeast splicing pathway, particularly assembly of the spliceosome. (1986)
  37. 10.1126/science.2544026
  38. 10.1016/0092-8674(84)90466-5
  39. 10.1093/nar/9.1.133
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, 11:21 p.m.)
Indexed 1 year ago (Aug. 4, 2024, 11:14 a.m.)
Issued 34 years, 2 months ago (July 1, 1991)
Published 34 years, 2 months ago (July 1, 1991)
Published Online 34 years, 2 months ago (July 1, 1991)
Published Print 34 years, 2 months ago (July 1, 1991)
Funders 0

None

@article{Deshler_1991, title={Unexpected point mutations activate cryptic 3’ splice sites by perturbing a natural secondary structure within a yeast intron.}, volume={5}, ISSN={1549-5477}, url={http://dx.doi.org/10.1101/gad.5.7.1252}, DOI={10.1101/gad.5.7.1252}, number={7}, journal={Genes & Development}, publisher={Cold Spring Harbor Laboratory}, author={Deshler, J O and Rossi, J J}, year={1991}, month=jul, pages={1252–1263} }