Abstract
We have identified two classical extragenic suppressors, spp41 and spp42, of the temperature sensitive (ts) allele prp4-73. The prp4(+) gene of Schizosaccharomyces pombe encodes a protein kinase. Mutations in both suppressor genes suppress the growth and the pre-mRNA splicing defect of prp4-73(ts) at the restrictive temperature (36 degrees ). spp41 and spp42 are synthetically lethal with each other in the presence of prp4-73(ts), indicating a functional relationship between spp41 and spp42. The suppressor genes were mapped on the left arm of chromosome I proximal to the his6 gene. Based on our mapping data we isolated spp42 by screening PCR fragments for functional complementation of the prp4-73(ts) mutant at the restrictive temperature. spp42 encodes a large protein (p275), which is the homologue of Prp8p. This protein has been shown in budding yeast and mammalian cells to be a bona fide pre-mRNA splicing factor. Taken together with other recent genetic and biochemical data, our results suggest that Prp4 kinase plays an important role in the formation of catalytic spliceosomes.
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Selected References
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- Abovich N., Legrain P., Rosbash M. The yeast PRP6 gene encodes a U4/U6 small nuclear ribonucleoprotein particle (snRNP) protein, and the PRP9 gene encodes a protein required for U2 snRNP binding. Mol Cell Biol. 1990 Dec;10(12):6417–6425. doi: 10.1128/mcb.10.12.6417. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Alahari S. K., Schmidt H., Käufer N. F. The fission yeast prp4+ gene involved in pre-mRNA splicing codes for a predicted serine/threonine kinase and is essential for growth. Nucleic Acids Res. 1993 Aug 25;21(17):4079–4083. doi: 10.1093/nar/21.17.4079. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Barbet N., Muriel W. J., Carr A. M. Versatile shuttle vectors and genomic libraries for use with Schizosaccharomyces pombe. Gene. 1992 May 1;114(1):59–66. doi: 10.1016/0378-1119(92)90707-v. [DOI] [PubMed] [Google Scholar]
- Duncan P. I., Stojdl D. F., Marius R. M., Scheit K. H., Bell J. C. The Clk2 and Clk3 dual-specificity protein kinases regulate the intranuclear distribution of SR proteins and influence pre-mRNA splicing. Exp Cell Res. 1998 Jun 15;241(2):300–308. doi: 10.1006/excr.1998.4083. [DOI] [PubMed] [Google Scholar]
- Gatermann K. B., Hoffmann A., Rosenberg G. H., Käufer N. F. Introduction of functional artificial introns into the naturally intronless ura4 gene of Schizosaccharomyces pombe. Mol Cell Biol. 1989 Apr;9(4):1526–1535. doi: 10.1128/mcb.9.4.1526. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gross T., Lützelberger M., Weigmann H., Klingenhoff A., Shenoy S., Käufer N. F. 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 Mar 1;25(5):1028–1035. doi: 10.1093/nar/25.5.1028. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gross T., Richert K., Mierke C., Lützelberger M., Käufer N. F. Identification and characterization of srp1, a gene of fission yeast encoding a RNA binding domain and a RS domain typical of SR splicing factors. Nucleic Acids Res. 1998 Jan 15;26(2):505–511. doi: 10.1093/nar/26.2.505. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gui J. F., Lane W. S., Fu X. D. A serine kinase regulates intracellular localization of splicing factors in the cell cycle. Nature. 1994 Jun 23;369(6482):678–682. doi: 10.1038/369678a0. [DOI] [PubMed] [Google Scholar]
- Guthrie C. Messenger RNA splicing in yeast: clues to why the spliceosome is a ribonucleoprotein. Science. 1991 Jul 12;253(5016):157–163. doi: 10.1126/science.1853200. [DOI] [PubMed] [Google Scholar]
- Habara Y., Urushiyama S., Tani T., Ohshima Y. The fission yeast prp10(+) gene involved in pre-mRNA splicing encodes a homologue of highly conserved splicing factor, SAP155. Nucleic Acids Res. 1998 Dec 15;26(24):5662–5669. doi: 10.1093/nar/26.24.5662. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hodges P. E., Jackson S. P., Brown J. D., Beggs J. D. Extraordinary sequence conservation of the PRP8 splicing factor. Yeast. 1995 Apr 15;11(4):337–342. doi: 10.1002/yea.320110406. [DOI] [PubMed] [Google Scholar]
- Käufer N. F., Simanis V., Nurse P. Fission yeast Schizosaccharomyces pombe correctly excises a mammalian RNA transcript intervening sequence. Nature. 1985 Nov 7;318(6041):78–80. doi: 10.1038/318078a0. [DOI] [PubMed] [Google Scholar]
- Lundgren K., Allan S., Urushiyama S., Tani T., Ohshima Y., Frendewey D., Beach D. A connection between pre-mRNA splicing and the cell cycle in fission yeast: cdc28+ is allelic with prp8+ and encodes an RNA-dependent ATPase/helicase. Mol Biol Cell. 1996 Jul;7(7):1083–1094. doi: 10.1091/mbc.7.7.1083. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lücke S., Klöckner T., Palfi Z., Boshart M., Bindereif A. Trans mRNA splicing in trypanosomes: cloning and analysis of a PRP8-homologous gene from Trypanosoma brucei provides evidence for a U5-analogous RNP. EMBO J. 1997 Jul 16;16(14):4433–4440. doi: 10.1093/emboj/16.14.4433. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Manley J. L., Tacke R. SR proteins and splicing control. Genes Dev. 1996 Jul 1;10(13):1569–1579. doi: 10.1101/gad.10.13.1569. [DOI] [PubMed] [Google Scholar]
- Maundrell K. Thiamine-repressible expression vectors pREP and pRIP for fission yeast. Gene. 1993 Jan 15;123(1):127–130. doi: 10.1016/0378-1119(93)90551-d. [DOI] [PubMed] [Google Scholar]
- Moreno S., Klar A., Nurse P. Molecular genetic analysis of fission yeast Schizosaccharomyces pombe. Methods Enzymol. 1991;194:795–823. doi: 10.1016/0076-6879(91)94059-l. [DOI] [PubMed] [Google Scholar]
- Nischt R., Thüroff E., Küfer N. F. Molecular cloning of a ribosomal protein gene from the fission yeast Schizosaccharomyces pombe. Curr Genet. 1986;10(5):365–370. doi: 10.1007/BF00418408. [DOI] [PubMed] [Google Scholar]
- Ohkura H., Kinoshita N., Miyatani S., Toda T., Yanagida M. The fission yeast dis2+ gene required for chromosome disjoining encodes one of two putative type 1 protein phosphatases. Cell. 1989 Jun 16;57(6):997–1007. doi: 10.1016/0092-8674(89)90338-3. [DOI] [PubMed] [Google Scholar]
- Potashkin J., Li R., Frendewey D. Pre-mRNA splicing mutants of Schizosaccharomyces pombe. EMBO J. 1989 Feb;8(2):551–559. doi: 10.1002/j.1460-2075.1989.tb03409.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Potashkin J., Naik K., Wentz-Hunter K. U2AF homolog required for splicing in vivo. Science. 1993 Oct 22;262(5133):573–575. doi: 10.1126/science.8211184. [DOI] [PubMed] [Google Scholar]
- Prabhala G., Rosenberg G. H., Käufer N. F. Architectural features of pre-mRNA introns in the fission yeast Schizosaccharomyces pombe. Yeast. 1992 Mar;8(3):171–182. doi: 10.1002/yea.320080303. [DOI] [PubMed] [Google Scholar]
- Reyes J. L., Gustafson E. H., Luo H. R., Moore M. J., Konarska M. M. The C-terminal region of hPrp8 interacts with the conserved GU dinucleotide at the 5' splice site. RNA. 1999 Feb;5(2):167–179. doi: 10.1017/s1355838299981785. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Rosenberg G. H., Alahari S. K., Käufer N. F. prp4 from Schizosaccharomyces pombe, a mutant deficient in pre-mRNA splicing isolated using genes containing artificial introns. Mol Gen Genet. 1991 Apr;226(1-2):305–309. doi: 10.1007/BF00273617. [DOI] [PubMed] [Google Scholar]
- Ruby S. W., Abelson J. Pre-mRNA splicing in yeast. Trends Genet. 1991 Mar;7(3):79–85. doi: 10.1016/0168-9525(91)90276-V. [DOI] [PubMed] [Google Scholar]
- Schmidt H. Effective long range mapping in Schizosaccharomyces pombe with the help of swi5. Curr Genet. 1993 Sep;24(3):271–273. doi: 10.1007/BF00351803. [DOI] [PubMed] [Google Scholar]
- Sha M., Levy T., Kois P., Konarska M. M. Probing of the spliceosome with site-specifically derivatized 5' splice site RNA oligonucleotides. RNA. 1998 Sep;4(9):1069–1082. doi: 10.1017/s1355838298980682. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Staley J. P., Guthrie C. Mechanical devices of the spliceosome: motors, clocks, springs, and things. Cell. 1998 Feb 6;92(3):315–326. doi: 10.1016/s0092-8674(00)80925-3. [DOI] [PubMed] [Google Scholar]
- Teigelkamp S., Newman A. J., Beggs J. D. Extensive interactions of PRP8 protein with the 5' and 3' splice sites during splicing suggest a role in stabilization of exon alignment by U5 snRNA. EMBO J. 1995 Jun 1;14(11):2602–2612. doi: 10.1002/j.1460-2075.1995.tb07258.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Umen J. G., Guthrie C. Mutagenesis of the yeast gene PRP8 reveals domains governing the specificity and fidelity of 3' splice site selection. Genetics. 1996 Jun;143(2):723–739. doi: 10.1093/genetics/143.2.723. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Urushiyama S., Tani T., Ohshima Y. Isolation of novel pre-mRNA splicing mutants of Schizosaccharomyces pombe. Mol Gen Genet. 1996 Nov 27;253(1-2):118–127. doi: 10.1007/s004380050304. [DOI] [PubMed] [Google Scholar]
- 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 Sep;147(1):101–115. doi: 10.1093/genetics/147.1.101. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Wang J., Manley J. L. Regulation of pre-mRNA splicing in metazoa. Curr Opin Genet Dev. 1997 Apr;7(2):205–211. doi: 10.1016/s0959-437x(97)80130-x. [DOI] [PubMed] [Google Scholar]
- Will C. L., Lührmann R. Protein functions in pre-mRNA splicing. Curr Opin Cell Biol. 1997 Jun;9(3):320–328. doi: 10.1016/s0955-0674(97)80003-8. [DOI] [PubMed] [Google Scholar]