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. 2002 Jul;161(3):1029–1042. doi: 10.1093/genetics/161.3.1029

The Saccharomyces cerevisiae RNase mitochondrial RNA processing is critical for cell cycle progression at the end of mitosis.

Ti Cai 1, Jason Aulds 1, Tina Gill 1, Michael Cerio 1, Mark E Schmitt 1
PMCID: PMC1462176  PMID: 12136008

Abstract

We have identified a cell cycle delay in Saccharomyces cerevisiae RNase MRP mutants. Mutants delay with large budded cells, dumbbell-shaped nuclei, and extended spindles characteristic of "exit from mitosis" mutants. In accord with this, a RNase MRP mutation can be suppressed by overexpressing the polo-like kinase CDC5 or by deleting the B-type cyclin CLB1, without restoring the MRP-dependent rRNA-processing step. In addition, we identified a series of genetic interactions between RNase MRP mutations and mutations in CDC5, CDC14, CDC15, CLB2, and CLB5. As in most "exit from mitosis" mutants, levels of the Clb2 cyclin were increased. The buildup of Clb2 protein is not the result of a defect in the release of the Cdc14 phosphatase from the nucleolus, but rather the result of an increase in CLB2 mRNA levels. These results indicate a clear role of RNase MRP in cell cycle progression at the end of mitosis. Conservation of this function in humans may explain many of the pleiotropic phenotypes of cartilage hair hypoplasia.

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Selected References

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  1. Cai T., Reilly T. R., Cerio M., Schmitt M. E. Mutagenesis of SNM1, which encodes a protein component of the yeast RNase MRP, reveals a role for this ribonucleoprotein endoribonuclease in plasmid segregation. Mol Cell Biol. 1999 Nov;19(11):7857–7869. doi: 10.1128/mcb.19.11.7857. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Cai T., Schmitt M. E. Characterization of ribonuclease MRP function. Methods Enzymol. 2001;342:135–142. doi: 10.1016/s0076-6879(01)42541-9. [DOI] [PubMed] [Google Scholar]
  3. Carlson M., Botstein D. Two differentially regulated mRNAs with different 5' ends encode secreted with intracellular forms of yeast invertase. Cell. 1982 Jan;28(1):145–154. doi: 10.1016/0092-8674(82)90384-1. [DOI] [PubMed] [Google Scholar]
  4. Chamberlain J. R., Lee Y., Lane W. S., Engelke D. R. Purification and characterization of the nuclear RNase P holoenzyme complex reveals extensive subunit overlap with RNase MRP. Genes Dev. 1998 Jun 1;12(11):1678–1690. doi: 10.1101/gad.12.11.1678. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Chang D. D., Clayton D. A. A novel endoribonuclease cleaves at a priming site of mouse mitochondrial DNA replication. EMBO J. 1987 Feb;6(2):409–417. doi: 10.1002/j.1460-2075.1987.tb04770.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Charles J. F., Jaspersen S. L., Tinker-Kulberg R. L., Hwang L., Szidon A., Morgan D. O. The Polo-related kinase Cdc5 activates and is destroyed by the mitotic cyclin destruction machinery in S. cerevisiae. Curr Biol. 1998 Apr 23;8(9):497–507. doi: 10.1016/s0960-9822(98)70201-5. [DOI] [PubMed] [Google Scholar]
  7. Chu S., Archer R. H., Zengel J. M., Lindahl L. The RNA of RNase MRP is required for normal processing of ribosomal RNA. Proc Natl Acad Sci U S A. 1994 Jan 18;91(2):659–663. doi: 10.1073/pnas.91.2.659. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Chu S., Zengel J. M., Lindahl L. A novel protein shared by RNase MRP and RNase P. RNA. 1997 Apr;3(4):382–391. [PMC free article] [PubMed] [Google Scholar]
  9. Clayton D. A. A big development for a small RNA. Nature. 2001 Mar 1;410(6824):29–31. doi: 10.1038/35065191. [DOI] [PubMed] [Google Scholar]
  10. Cross Frederick R., Archambault Vincent, Miller Mary, Klovstad Martha. Testing a mathematical model of the yeast cell cycle. Mol Biol Cell. 2002 Jan;13(1):52–70. doi: 10.1091/mbc.01-05-0265. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Dichtl B., Tollervey D. Pop3p is essential for the activity of the RNase MRP and RNase P ribonucleoproteins in vivo. EMBO J. 1997 Jan 15;16(2):417–429. doi: 10.1093/emboj/16.2.417. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Fitch I., Dahmann C., Surana U., Amon A., Nasmyth K., Goetsch L., Byers B., Futcher B. Characterization of four B-type cyclin genes of the budding yeast Saccharomyces cerevisiae. Mol Biol Cell. 1992 Jul;3(7):805–818. doi: 10.1091/mbc.3.7.805. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Henry Y., Wood H., Morrissey J. P., Petfalski E., Kearsey S., Tollervey D. The 5' end of yeast 5.8S rRNA is generated by exonucleases from an upstream cleavage site. EMBO J. 1994 May 15;13(10):2452–2463. doi: 10.1002/j.1460-2075.1994.tb06530.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Jaspersen S. L., Charles J. F., Tinker-Kulberg R. L., Morgan D. O. A late mitotic regulatory network controlling cyclin destruction in Saccharomyces cerevisiae. Mol Biol Cell. 1998 Oct;9(10):2803–2817. doi: 10.1091/mbc.9.10.2803. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Juvonen E., Mäkitie O., Mäkipernaa A., Ruutu T., Kaitila I., Rajantie J. Defective in-vitro colony formation of haematopoietic progenitors in patients with cartilage-hair hypoplasia and history of anaemia. Eur J Pediatr. 1995 Jan;154(1):30–34. doi: 10.1007/BF01972969. [DOI] [PubMed] [Google Scholar]
  16. Longtine M. S., McKenzie A., 3rd, Demarini D. J., Shah N. G., Wach A., Brachat A., Philippsen P., Pringle J. R. Additional modules for versatile and economical PCR-based gene deletion and modification in Saccharomyces cerevisiae. Yeast. 1998 Jul;14(10):953–961. doi: 10.1002/(SICI)1097-0061(199807)14:10<953::AID-YEA293>3.0.CO;2-U. [DOI] [PubMed] [Google Scholar]
  17. Lygerou Z., Mitchell P., Petfalski E., Séraphin B., Tollervey D. The POP1 gene encodes a protein component common to the RNase MRP and RNase P ribonucleoproteins. Genes Dev. 1994 Jun 15;8(12):1423–1433. doi: 10.1101/gad.8.12.1423. [DOI] [PubMed] [Google Scholar]
  18. Lygerou Z., Pluk H., van Venrooij W. J., Séraphin B. hPop1: an autoantigenic protein subunit shared by the human RNase P and RNase MRP ribonucleoproteins. EMBO J. 1996 Nov 1;15(21):5936–5948. [PMC free article] [PubMed] [Google Scholar]
  19. Mäkitie O., Kaitila I., Savilahti E. Susceptibility to infections and in vitro immune functions in cartilage-hair hypoplasia. Eur J Pediatr. 1998 Oct;157(10):816–820. doi: 10.1007/s004310050943. [DOI] [PubMed] [Google Scholar]
  20. Mäkitie O., Sulisalo T., de la Chapelle A., Kaitila I. Cartilage-hair hypoplasia. J Med Genet. 1995 Jan;32(1):39–43. doi: 10.1136/jmg.32.1.39. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Paluh J. L., Clayton D. A. A functional dominant mutation in Schizosaccharomyces pombe RNase MRP RNA affects nuclear RNA processing and requires the mitochondrial-associated nuclear mutation ptp1-1 for viability. EMBO J. 1996 Sep 2;15(17):4723–4733. [PMC free article] [PubMed] [Google Scholar]
  22. Pringle J. R., Adams A. E., Drubin D. G., Haarer B. K. Immunofluorescence methods for yeast. Methods Enzymol. 1991;194:565–602. doi: 10.1016/0076-6879(91)94043-c. [DOI] [PubMed] [Google Scholar]
  23. Reimer G., Raska I., Scheer U., Tan E. M. Immunolocalization of 7-2-ribonucleoprotein in the granular component of the nucleolus. Exp Cell Res. 1988 May;176(1):117–128. doi: 10.1016/0014-4827(88)90126-7. [DOI] [PubMed] [Google Scholar]
  24. Ridanpä M., van Eenennaam H., Pelin K., Chadwick R., Johnson C., Yuan B., vanVenrooij W., Pruijn G., Salmela R., Rockas S. Mutations in the RNA component of RNase MRP cause a pleiotropic human disease, cartilage-hair hypoplasia. Cell. 2001 Jan 26;104(2):195–203. doi: 10.1016/s0092-8674(01)00205-7. [DOI] [PubMed] [Google Scholar]
  25. Schmitt M. E., Brown T. A., Trumpower B. L. A rapid and simple method for preparation of RNA from Saccharomyces cerevisiae. Nucleic Acids Res. 1990 May 25;18(10):3091–3092. doi: 10.1093/nar/18.10.3091. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Schmitt M. E., Clayton D. A. Characterization of a unique protein component of yeast RNase MRP: an RNA-binding protein with a zinc-cluster domain. Genes Dev. 1994 Nov 1;8(21):2617–2628. doi: 10.1101/gad.8.21.2617. [DOI] [PubMed] [Google Scholar]
  27. Schmitt M. E., Clayton D. A. Nuclear RNase MRP is required for correct processing of pre-5.8S rRNA in Saccharomyces cerevisiae. Mol Cell Biol. 1993 Dec;13(12):7935–7941. doi: 10.1128/mcb.13.12.7935. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Schmitt M. E., Clayton D. A. Yeast site-specific ribonucleoprotein endoribonuclease MRP contains an RNA component homologous to mammalian RNase MRP RNA and essential for cell viability. Genes Dev. 1992 Oct;6(10):1975–1985. doi: 10.1101/gad.6.10.1975. [DOI] [PubMed] [Google Scholar]
  29. Shadel G. S., Buckenmeyer G. A., Clayton D. A., Schmitt M. E. Mutational analysis of the RNA component of Saccharomyces cerevisiae RNase MRP reveals distinct nuclear phenotypes. Gene. 2000 Mar 7;245(1):175–184. doi: 10.1016/s0378-1119(00)00013-5. [DOI] [PubMed] [Google Scholar]
  30. Shirayama M., Zachariae W., Ciosk R., Nasmyth K. The Polo-like kinase Cdc5p and the WD-repeat protein Cdc20p/fizzy are regulators and substrates of the anaphase promoting complex in Saccharomyces cerevisiae. EMBO J. 1998 Mar 2;17(5):1336–1349. doi: 10.1093/emboj/17.5.1336. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Shortle D., Haber J. E., Botstein D. Lethal disruption of the yeast actin gene by integrative DNA transformation. Science. 1982 Jul 23;217(4557):371–373. doi: 10.1126/science.7046050. [DOI] [PubMed] [Google Scholar]
  32. Shou W., Seol J. H., Shevchenko A., Baskerville C., Moazed D., Chen Z. W., Jang J., Shevchenko A., Charbonneau H., Deshaies R. J. Exit from mitosis is triggered by Tem1-dependent release of the protein phosphatase Cdc14 from nucleolar RENT complex. Cell. 1999 Apr 16;97(2):233–244. doi: 10.1016/s0092-8674(00)80733-3. [DOI] [PubMed] [Google Scholar]
  33. Shu Y., Yang H., Hallberg E., Hallberg R. Molecular genetic analysis of Rts1p, a B' regulatory subunit of Saccharomyces cerevisiae protein phosphatase 2A. Mol Cell Biol. 1997 Jun;17(6):3242–3253. doi: 10.1128/mcb.17.6.3242. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Sikorski R. S., Boeke J. D. In vitro mutagenesis and plasmid shuffling: from cloned gene to mutant yeast. Methods Enzymol. 1991;194:302–318. doi: 10.1016/0076-6879(91)94023-6. [DOI] [PubMed] [Google Scholar]
  35. Spellman P. T., Sherlock G., Zhang M. Q., Iyer V. R., Anders K., Eisen M. B., Brown P. O., Botstein D., Futcher B. Comprehensive identification of cell cycle-regulated genes of the yeast Saccharomyces cerevisiae by microarray hybridization. Mol Biol Cell. 1998 Dec;9(12):3273–3297. doi: 10.1091/mbc.9.12.3273. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Stolc V., Altman S. Rpp1, an essential protein subunit of nuclear RNase P required for processing of precursor tRNA and 35S precursor rRNA in Saccharomyces cerevisiae. Genes Dev. 1997 Sep 15;11(18):2414–2425. doi: 10.1101/gad.11.18.2414. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Surana U., Amon A., Dowzer C., McGrew J., Byers B., Nasmyth K. Destruction of the CDC28/CLB mitotic kinase is not required for the metaphase to anaphase transition in budding yeast. EMBO J. 1993 May;12(5):1969–1978. doi: 10.1002/j.1460-2075.1993.tb05846.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Visintin R., Craig K., Hwang E. S., Prinz S., Tyers M., Amon A. The phosphatase Cdc14 triggers mitotic exit by reversal of Cdk-dependent phosphorylation. Mol Cell. 1998 Dec;2(6):709–718. doi: 10.1016/s1097-2765(00)80286-5. [DOI] [PubMed] [Google Scholar]
  39. Visintin R., Hwang E. S., Amon A. Cfi1 prevents premature exit from mitosis by anchoring Cdc14 phosphatase in the nucleolus. Nature. 1999 Apr 29;398(6730):818–823. doi: 10.1038/19775. [DOI] [PubMed] [Google Scholar]

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