The N-terminus of Prp1 (Prp6/U5-102 K) is Essential for Spliceosome Activation in Vivo

Overview

Journal Nucleic Acids Res

Publisher Oxford University Press

Specialty Biochemistry

Date 2009 Dec 17

PMID 20007600

Citations 8

Authors

Martin Lutzelberger

Claudia A Bottner

Wiebke Schwelnus

Susanne Zock-Emmenthal

Aleh Razanau

Norbert F Kaufer

Affiliations

Soon will be listed here.

Abstract

The spliceosomal protein Prp1 (Prp6/U5-102 K) is necessary for the integrity of pre-catalytic spliceosomal complexes. We have identified a novel regulatory function for Prp1. Expression of mutations in the N-terminus of Prp1 leads to the accumulation of pre-catalytic spliceosomal complexes containing the five snRNAs U1, U2, U5 and U4/U6 and pre-mRNAs. The mutations in the N-terminus, which prevent splicing to occur, include in vitro and in vivo identified phosphorylation sites of Prp4 kinase. These sites are highly conserved in the human ortholog U5-102 K. The results presented here demonstrate that structural integrity of the N-terminus is required to mediate a splicing event, but is not necessary for the assembly of spliceosomes.

Citing Articles

An Allosteric Network for Spliceosome Activation Revealed by High-Throughput Suppressor Analysis in .

Brow D Genetics. 2019; 212(1):111-124.

PMID: 30898770 PMC: 6499515. DOI: 10.1534/genetics.119.301922.

Salinity-Induced Palmella Formation Mechanism in Halotolerant Algae Revealed by Quantitative Proteomics and Phosphoproteomics.

Wei S, Bian Y, Zhao Q, Chen S, Mao J, Song C Front Plant Sci. 2017; 8:810.

PMID: 28588593 PMC: 5441111. DOI: 10.3389/fpls.2017.00810.

Prp4 Kinase Grants the License to Splice: Control of Weak Splice Sites during Spliceosome Activation.

Eckert D, Andree N, Razanau A, Zock-Emmenthal S, Lutzelberger M, Plath S PLoS Genet. 2016; 12(1):e1005768.

PMID: 26730850 PMC: 4701394. DOI: 10.1371/journal.pgen.1005768.

Structural and functional characterization of the N terminus of Schizosaccharomyces pombe Cwf10.

Livesay S, Collier S, Bitton D, Bahler J, Ohi M Eukaryot Cell. 2013; 12(11):1472-89.

PMID: 24014766 PMC: 3837936. DOI: 10.1128/EC.00140-13.

Splicing functions and global dependency on fission yeast slu7 reveal diversity in spliceosome assembly.

Banerjee S, Khandelia P, Melangath G, Bashir S, Nagampalli V, Vijayraghavan U Mol Cell Biol. 2013; 33(16):3125-36.

PMID: 23754748 PMC: 3753915. DOI: 10.1128/MCB.00007-13.

References

Liu S, Li P, Dybkov O, Nottrott S, Hartmuth K, Luhrmann R . Binding of the human Prp31 Nop domain to a composite RNA-protein platform in U4 snRNP. Science. 2007; 316(5821):115-20. DOI: 10.1126/science.1137924. View

Ohi M, Ren L, Wall J, Gould K, Walz T . Structural characterization of the fission yeast U5.U2/U6 spliceosome complex. Proc Natl Acad Sci U S A. 2007; 104(9):3195-200. PMC: 1805518. DOI: 10.1073/pnas.0611591104. View

Deckert J, Hartmuth K, Boehringer D, Behzadnia N, Will C, Kastner B . Protein composition and electron microscopy structure of affinity-purified human spliceosomal B complexes isolated under physiological conditions. Mol Cell Biol. 2006; 26(14):5528-43. PMC: 1592722. DOI: 10.1128/MCB.00582-06. View

Sato M, Dhut S, Toda T . New drug-resistant cassettes for gene disruption and epitope tagging in Schizosaccharomyces pombe. Yeast. 2005; 22(7):583-91. DOI: 10.1002/yea.1233. View

Makarov E, Makarova O, Achsel T, Luhrmann R . The human homologue of the yeast splicing factor prp6p contains multiple TPR elements and is stably associated with the U5 snRNP via protein-protein interactions. J Mol Biol. 2000; 298(4):567-75. DOI: 10.1006/jmbi.2000.3685. View

Wahl M, Will C, Luhrmann R . The spliceosome: design principles of a dynamic RNP machine. Cell. 2009; 136(4):701-18. DOI: 10.1016/j.cell.2009.02.009. View

Gross T, Lutzelberger M, Weigmann H, Klingenhoff A, Shenoy S, Kaufer N . Functional analysis of the fission yeast Prp4 protein kinase involved in pre-mRNA splicing and isolation of a putative mammalian homologue. Nucleic Acids Res. 1997; 25(5):1028-35. PMC: 146536. DOI: 10.1093/nar/25.5.1028. View

Bon E, Casaregola S, Blandin G, Llorente B, Neuveglise C, Munsterkotter M . Molecular evolution of eukaryotic genomes: hemiascomycetous yeast spliceosomal introns. Nucleic Acids Res. 2003; 31(4):1121-35. PMC: 150231. DOI: 10.1093/nar/gkg213. View

Listerman I, Sapra A, Neugebauer K . Cotranscriptional coupling of splicing factor recruitment and precursor messenger RNA splicing in mammalian cells. Nat Struct Mol Biol. 2006; 13(9):815-22. DOI: 10.1038/nsmb1135. View

10.

Brow D . Allosteric cascade of spliceosome activation. Annu Rev Genet. 2002; 36:333-60. DOI: 10.1146/annurev.genet.36.043002.091635. View

11.

Maundrell K . Thiamine-repressible expression vectors pREP and pRIP for fission yeast. Gene. 1993; 123(1):127-30. DOI: 10.1016/0378-1119(93)90551-d. View

12.

Rigaut G, Shevchenko A, Rutz B, Wilm M, Mann M, Seraphin B . A generic protein purification method for protein complex characterization and proteome exploration. Nat Biotechnol. 1999; 17(10):1030-2. DOI: 10.1038/13732. View

13.

Potashkin J, Kim D, Fons M, Humphrey T, Frendewey D . Cell-division-cycle defects associated with fission yeast pre-mRNA splicing mutants. Curr Genet. 1998; 34(3):153-63. DOI: 10.1007/s002940050381. View

14.

Dellaire G, Makarov E, Cowger J, Longman D, Sutherland H, Luhrmann R . Mammalian PRP4 kinase copurifies and interacts with components of both the U5 snRNP and the N-CoR deacetylase complexes. Mol Cell Biol. 2002; 22(14):5141-56. PMC: 139773. DOI: 10.1128/MCB.22.14.5141-5156.2002. View

15.

Ohi M, Vander Kooi C, Rosenberg J, Ren L, Hirsch J, Chazin W . Structural and functional analysis of essential pre-mRNA splicing factor Prp19p. Mol Cell Biol. 2004; 25(1):451-60. PMC: 538785. DOI: 10.1128/MCB.25.1.451-460.2005. View

16.

Urushiyama S, Tani T, Ohshima Y . The prp1+ gene required for pre-mRNA splicing in Schizosaccharomyces pombe encodes a protein that contains TPR motifs and is similar to Prp6p of budding yeast. Genetics. 1997; 147(1):101-15. PMC: 1208094. DOI: 10.1093/genetics/147.1.101. View

17.

Bahler J, Wu J, Longtine M, Shah N, McKenzie 3rd A, Steever A . Heterologous modules for efficient and versatile PCR-based gene targeting in Schizosaccharomyces pombe. Yeast. 1998; 14(10):943-51. DOI: 10.1002/(SICI)1097-0061(199807)14:10<943::AID-YEA292>3.0.CO;2-Y. View

18.

Tardiff D, Lacadie S, Rosbash M . A genome-wide analysis indicates that yeast pre-mRNA splicing is predominantly posttranscriptional. Mol Cell. 2006; 24(6):917-29. PMC: 1828117. DOI: 10.1016/j.molcel.2006.12.002. View

19.

Newo A, Lutzelberger M, Bottner C, Wehland J, Wissing J, Jansch L . Proteomic analysis of the U1 snRNP of Schizosaccharomyces pombe reveals three essential organism-specific proteins. Nucleic Acids Res. 2007; 35(5):1391-401. PMC: 1865046. DOI: 10.1093/nar/gkl1144. View

20.

Keeney J, Boeke J . Efficient targeted integration at leu1-32 and ura4-294 in Schizosaccharomyces pombe. Genetics. 1994; 136(3):849-56. PMC: 1205890. DOI: 10.1093/genetics/136.3.849. View