Partial Purification of the Yeast U2 SnRNP Reveals a Novel Yeast Pre-mRNA Splicing Factor Required for Pre-spliceosome Assembly

Overview

Journal EMBO J

Specialties Biotechnology
Molecular Biology

Date 1999 Jun 16

PMID 10369685

Citations 27

Authors

F Caspary

A Shevchenko

M Wilm

B Seraphin

Affiliations

Soon will be listed here.

Abstract

We have partially purified the U2 snRNP of Saccharomyces cerevisiae. Identification of some proteins consistently found in the purified fractions by nanoelectrospray mass spectrometry indicated the presence of a novel splicing factor named Rse1p. The RSE1 gene is essential and codes for a 148.2 kDa protein. We demonstrated that Rse1p associates specifically with U2 snRNA at low salt concentrations. In addition, we showed that Rse1p is a component of the pre-spliceosome. Depletion of Rse1p and analysis of a conditional mutant indicated that Rse1p was required for efficient splicing in vivo. In vitro Rse1p is required for the formation of pre-spliceosomes. Database searches revealed that Rse1p is conserved in humans and that it belongs to a large protein family that includes polyadenylation factors and DNA repair proteins. The characteristics of Rse1p suggest that its human homologue could be a subunit of the SF3 splicing factor.

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References

Abovich N, Rosbash M . Cross-intron bridging interactions in the yeast commitment complex are conserved in mammals. Cell. 1997; 89(3):403-12. DOI: 10.1016/s0092-8674(00)80221-4. View

Will C, Luhrmann R . Protein functions in pre-mRNA splicing. Curr Opin Cell Biol. 1997; 9(3):320-8. DOI: 10.1016/s0955-0674(97)80003-8. View

Burd C, Dreyfuss G . Conserved structures and diversity of functions of RNA-binding proteins. Science. 1994; 265(5172):615-21. DOI: 10.1126/science.8036511. View

Hodges P, Beggs J . RNA splicing. U2 fulfils a commitment. Curr Biol. 1994; 4(3):264-7. DOI: 10.1016/s0960-9822(00)00061-0. View

Abovich N, Liao X, Rosbash M . The yeast MUD2 protein: an interaction with PRP11 defines a bridge between commitment complexes and U2 snRNP addition. Genes Dev. 1994; 8(7):843-54. DOI: 10.1101/gad.8.7.843. View

REED R . The prespliceosome components SAP 49 and SAP 145 interact in a complex implicated in tethering U2 snRNP to the branch site. Genes Dev. 1994; 8(16):1974-83. DOI: 10.1101/gad.8.16.1974. View

Rain J, Legrain P . Toward a functional analysis of the yeast genome through exhaustive two-hybrid screens. Nat Genet. 1997; 16(3):277-82. DOI: 10.1038/ng0797-277. View

Thompson J, Gibson T, Plewniak F, Jeanmougin F, Higgins D . The CLUSTAL_X windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools. Nucleic Acids Res. 1998; 25(24):4876-82. PMC: 147148. DOI: 10.1093/nar/25.24.4876. View

IGEL H, Wells S, Perriman R, Ares Jr M . Conservation of structure and subunit interactions in yeast homologues of splicing factor 3b (SF3b) subunits. RNA. 1998; 4(1):1-10. PMC: 1369591. View

10.

Knecht S, Kramer A . Molecular characterization of a novel, widespread nuclear protein that colocalizes with spliceosome components. Mol Biol Cell. 1998; 9(1):143-60. PMC: 25229. DOI: 10.1091/mbc.9.1.143. View

11.

Staley J, Guthrie C . Mechanical devices of the spliceosome: motors, clocks, springs, and things. Cell. 1998; 92(3):315-26. DOI: 10.1016/s0092-8674(00)80925-3. View

12.

Berglund J, Abovich N, Rosbash M . A cooperative interaction between U2AF65 and mBBP/SF1 facilitates branchpoint region recognition. Genes Dev. 1998; 12(6):858-67. PMC: 316625. DOI: 10.1101/gad.12.6.858. View

13.

Wang C, Chua K, Seghezzi W, Lees E, Gozani O, REED R . Phosphorylation of spliceosomal protein SAP 155 coupled with splicing catalysis. Genes Dev. 1998; 12(10):1409-14. PMC: 316838. DOI: 10.1101/gad.12.10.1409. View

14.

Gottschalk A, Tang J, Puig O, Salgado J, Neubauer G, Colot H . A comprehensive biochemical and genetic analysis of the yeast U1 snRNP reveals five novel proteins. RNA. 1998; 4(4):374-93. PMC: 1369625. View

15.

Gozani O, Potashkin J, REED R . A potential role for U2AF-SAP 155 interactions in recruiting U2 snRNP to the branch site. Mol Cell Biol. 1998; 18(8):4752-60. PMC: 109061. DOI: 10.1128/MCB.18.8.4752. View

16.

Puig O, Rutz B, Luukkonen B, Seraphin B . New constructs and strategies for efficient PCR-based gene manipulations in yeast. Yeast. 1998; 14(12):1139-46. DOI: 10.1002/(SICI)1097-0061(19980915)14:12<1139::AID-YEA306>3.0.CO;2-B. View

17.

Caspary F, Seraphin B . The yeast U2A'/U2B complex is required for pre-spliceosome formation. EMBO J. 1998; 17(21):6348-58. PMC: 1170959. DOI: 10.1093/emboj/17.21.6348. View

18.

Chen E, Frand A, Chitouras E, Kaiser C . A link between secretion and pre-mRNA processing defects in Saccharomyces cerevisiae and the identification of a novel splicing gene, RSE1. Mol Cell Biol. 1998; 18(12):7139-46. PMC: 109295. DOI: 10.1128/MCB.18.12.7139. View

19.

Seraphin B . Sm and Sm-like proteins assemble in two related complexes of deep evolutionary origin. EMBO J. 1999; 18(12):3451-62. PMC: 1171424. DOI: 10.1093/emboj/18.12.3451. View

20.

Soni R, Carmichael J, Murray J . Parameters affecting lithium acetate-mediated transformation of Saccharomyces cerevisiae and development of a rapid and simplified procedure. Curr Genet. 1993; 24(5):455-9. DOI: 10.1007/BF00351857. View