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. 1990 Jul;10(7):3524–3534. doi: 10.1128/mcb.10.7.3524

Differential distribution of factors involved in pre-mRNA processing in the yeast cell nucleus.

J A Potashkin 1, R J Derby 1, D L Spector 1
PMCID: PMC360787  PMID: 2192258

Abstract

The yeast cell nucleus has previously been shown to be divided into two regions by a variety of microscopic approaches. We used antibodies specific for the 2,2,7-trimethylguanosine cap structure of small nuclear ribonucleic acids (snRNAs) and for a protein component of small nuclear ribonucleoprotein particles to identify the distribution of small nuclear ribonucleoprotein particles within the yeast cell nucleus. These studies were performed with the fission yeast Schizosaccharomyces pombe and the budding yeast Saccharomyces cerevisiae. By using immunofluorescence microscopy and immunoelectron microscopy, most of the abundant snRNAs were localized to the portion of the nucleus which has heretofore been referred to as the nucleolus. This distribution of snRNAs is different from that found in mammalian cells and suggests that the nucleolar portion of the yeast nucleus contains functional domains in addition to those associated with RNA polymerase I activity.

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Selected References

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  1. Aris J. P., Blobel G. Identification and characterization of a yeast nucleolar protein that is similar to a rat liver nucleolar protein. J Cell Biol. 1988 Jul;107(1):17–31. doi: 10.1083/jcb.107.1.17. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bourgeois C. A., Hernandez-Verdun D., Hubert J., Bouteille M. Silver staining of NORs in electron microscopy. Exp Cell Res. 1979 Oct 15;123(2):449–452. doi: 10.1016/0014-4827(79)90498-1. [DOI] [PubMed] [Google Scholar]
  3. Brennwald P., Porter G., Wise J. A. U2 small nuclear RNA is remarkably conserved between Schizosaccharomyces pombe and mammals. Mol Cell Biol. 1988 Dec;8(12):5575–5580. doi: 10.1128/mcb.8.12.5575. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Busch H., Reddy R., Rothblum L., Choi Y. C. SnRNAs, SnRNPs, and RNA processing. Annu Rev Biochem. 1982;51:617–654. doi: 10.1146/annurev.bi.51.070182.003153. [DOI] [PubMed] [Google Scholar]
  5. Chang T. H., Clark M. W., Lustig A. J., Cusick M. E., Abelson J. RNA11 protein is associated with the yeast spliceosome and is localized in the periphery of the cell nucleus. Mol Cell Biol. 1988 Jun;8(6):2379–2393. doi: 10.1128/mcb.8.6.2379. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Daskal Y., Smetana K., Busch H. Evidence from studies on segregated nucleoli that nucleolar silver staining proteins C23 and B23 are in the fibrillar component. Exp Cell Res. 1980 Jun;127(2):285–291. doi: 10.1016/0014-4827(80)90434-6. [DOI] [PubMed] [Google Scholar]
  7. Dimova R. N., Markov D. V., Gajdardjieva K. C., Dabeva M. D., Hadjiolov A. A. Electron microscopic localization of silver staining NOR-proteins in rat liver nucleoli upon D-galactosamine block of transcription. Eur J Cell Biol. 1982 Oct;28(2):272–277. [PubMed] [Google Scholar]
  8. Epstein P., Reddy R., Busch H. Multiple states of U3 RNA in Novikoff hepatoma nucleoli. Biochemistry. 1984 Nov 6;23(23):5421–5425. doi: 10.1021/bi00318a007. [DOI] [PubMed] [Google Scholar]
  9. Fakan S., Hernandez-Verdun D. The nucleolus and the nucleolar organizer regions. Biol Cell. 1986;56(3):189–205. doi: 10.1111/j.1768-322x.1986.tb00452.x. [DOI] [PubMed] [Google Scholar]
  10. Fu X. D., Maniatis T. Factor required for mammalian spliceosome assembly is localized to discrete regions in the nucleus. Nature. 1990 Feb 1;343(6257):437–441. doi: 10.1038/343437a0. [DOI] [PubMed] [Google Scholar]
  11. Goessens G. Nucleolar structure. Int Rev Cytol. 1984;87:107–158. doi: 10.1016/s0074-7696(08)62441-9. [DOI] [PubMed] [Google Scholar]
  12. Gordon C. N. Chromatin behaviour during the mitotic cell cycle of Saccharomyces cerevisiae. J Cell Sci. 1977 Apr;24:81–93. doi: 10.1242/jcs.24.1.81. [DOI] [PubMed] [Google Scholar]
  13. Hansmann I., Gebauer J., Bihl L., Grimm T. Onset of nucleolus organizer activity in early mouse embryogenesis and evidence for its regulation. Exp Cell Res. 1978 Jul;114(2):263–268. doi: 10.1016/0014-4827(78)90482-2. [DOI] [PubMed] [Google Scholar]
  14. Hernandez-Verdun D., Derenzini M., Bouteille M. The morphological relationship in electron microscopy between NOR-silver proteins and intranucleolar chromatin. Chromosoma. 1982;85(4):461–473. doi: 10.1007/BF00327343. [DOI] [PubMed] [Google Scholar]
  15. Hernandez-Verdun D., Hubert J., Bourgeois C. A., Bouteille M. Ultrastructural localization of Ag-NOR stained proteins in the nucleolus during the cell cycle and in other nucleolar structures. Chromosoma. 1980;79(3):349–362. doi: 10.1007/BF00327325. [DOI] [PubMed] [Google Scholar]
  16. Howell W. M., Denton T. E., Diamond J. R. Differential staining of the satellite regions of human acrocentric chromosomes. Experientia. 1975 Feb 15;31(2):260–262. doi: 10.1007/BF01990741. [DOI] [PubMed] [Google Scholar]
  17. Hubbell H. R., Rothblum L. I., Hsu T. C. Identification of a silver binding protein associated with the cytological silver staining of actively transcribing nucleolar regions. Cell Biol Int Rep. 1979 Oct;3(7):615–622. doi: 10.1016/0309-1651(79)90060-2. [DOI] [PubMed] [Google Scholar]
  18. Hubert J., Bureau J., Bouteille M. Anchorage of the nucleolus in the pore complex-lamina by a DNA-bearing structure masked in situ in rat liver nuclei. Biol Cell. 1984;52(2):91–101. doi: 10.1111/j.1768-322x.1985.tb00326.x. [DOI] [PubMed] [Google Scholar]
  19. Krainer A. R. Pre-mRNA splicing by complementation with purified human U1, U2, U4/U6 and U5 snRNPs. Nucleic Acids Res. 1988 Oct 25;16(20):9415–9429. doi: 10.1093/nar/16.20.9415. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Kubota T., Akatsuka T., Kanai Y. A monoclonal anti-double stranded DNA antibody from an autoimmune MRL/Mp-lpr/lpr mouse: specificity and idiotype in serum immunoglobulins. Immunol Lett. 1986 Nov 17;14(1):53–58. doi: 10.1016/0165-2478(86)90020-9. [DOI] [PubMed] [Google Scholar]
  21. Last R. L., Woolford J. L., Jr Identification and nuclear localization of yeast pre-messenger RNA processing components: RNA2 and RNA3 proteins. J Cell Biol. 1986 Dec;103(6 Pt 1):2103–2112. doi: 10.1083/jcb.103.6.2103. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Lerner E. A., Lerner M. R., Janeway C. A., Jr, Steitz J. A. Monoclonal antibodies to nucleic acid-containing cellular constituents: probes for molecular biology and autoimmune disease. Proc Natl Acad Sci U S A. 1981 May;78(5):2737–2741. doi: 10.1073/pnas.78.5.2737. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Lischwe M. A., Ochs R. L., Reddy R., Cook R. G., Yeoman L. C., Tan E. M., Reichlin M., Busch H. Purification and partial characterization of a nucleolar scleroderma antigen (Mr = 34,000; pI, 8.5) rich in NG,NG-dimethylarginine. J Biol Chem. 1985 Nov 15;260(26):14304–14310. [PubMed] [Google Scholar]
  24. McCully E. K., Robinow C. F. Mitosis in the fission yeast Schizosaccharomyces pombe: a comparative study with light and electron microscopy. J Cell Sci. 1971 Sep;9(2):475–507. doi: 10.1242/jcs.9.2.475. [DOI] [PubMed] [Google Scholar]
  25. Molenaar I., Sillevis Smitt W. W., Rozijn T. H., Tonino G. J. Biochemical and electron microscopic study of isolated yeast nuclei. Exp Cell Res. 1970 May;60(2):148–156. doi: 10.1016/0014-4827(70)90500-8. [DOI] [PubMed] [Google Scholar]
  26. Nyman U., Hallman H., Hadlaczky G., Pettersson I., Sharp G., Ringertz N. R. Intranuclear localization of snRNP antigens. J Cell Biol. 1986 Jan;102(1):137–144. doi: 10.1083/jcb.102.1.137. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Ochs R. L., Busch H. Further evidence that phosphoprotein C23 (110 kD/pI 5.1) is the nucleolar silver staining protein. Exp Cell Res. 1984 May;152(1):260–265. doi: 10.1016/0014-4827(84)90251-9. [DOI] [PubMed] [Google Scholar]
  28. Ochs R. L., Lischwe M. A., Shen E., Carroll R. E., Busch H. Nucleologenesis: composition and fate of prenucleolar bodies. Chromosoma. 1985;92(5):330–336. doi: 10.1007/BF00327463. [DOI] [PubMed] [Google Scholar]
  29. Ochs R. L., Lischwe M. A., Spohn W. H., Busch H. Fibrillarin: a new protein of the nucleolus identified by autoimmune sera. Biol Cell. 1985;54(2):123–133. doi: 10.1111/j.1768-322x.1985.tb00387.x. [DOI] [PubMed] [Google Scholar]
  30. Parker K. A., Steitz J. A. Structural analysis of the human U3 ribonucleoprotein particle reveal a conserved sequence available for base pairing with pre-rRNA. Mol Cell Biol. 1987 Aug;7(8):2899–2913. doi: 10.1128/mcb.7.8.2899. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Prestayko A. W., Tonato M., Busch H. Low molecular weight RNA associated with 28 s nucleolar RNA. J Mol Biol. 1970 Feb 14;47(3):505–515. doi: 10.1016/0022-2836(70)90318-9. [DOI] [PubMed] [Google Scholar]
  32. REYNOLDS E. S. The use of lead citrate at high pH as an electron-opaque stain in electron microscopy. J Cell Biol. 1963 Apr;17:208–212. doi: 10.1083/jcb.17.1.208. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Raman R., Sperling K. Patterns of silver staining on NORs of prematurely condensed muntjac chromosomes following RNA inhibition. Exp Cell Res. 1981 Oct;135(2):373–378. doi: 10.1016/0014-4827(81)90173-7. [DOI] [PubMed] [Google Scholar]
  34. Reimer G., Pollard K. M., Penning C. A., Ochs R. L., Lischwe M. A., Busch H., Tan E. M. Monoclonal autoantibody from a (New Zealand black x New Zealand white)F1 mouse and some human scleroderma sera target an Mr 34,000 nucleolar protein of the U3 RNP particle. Arthritis Rheum. 1987 Jul;30(7):793–800. doi: 10.1002/art.1780300709. [DOI] [PubMed] [Google Scholar]
  35. Reuter R., Appel B., Bringmann P., Rinke J., Lührmann R. 5'-Terminal caps of snRNAs are reactive with antibodies specific for 2,2,7-trimethylguanosine in whole cells and nuclear matrices. Double-label immunofluorescent studies with anti-m3G antibodies and with anti-RNP and anti-Sm autoantibodies. Exp Cell Res. 1984 Oct;154(2):548–560. doi: 10.1016/0014-4827(84)90179-4. [DOI] [PubMed] [Google Scholar]
  36. Riedel N., Wise J. A., Swerdlow H., Mak A., Guthrie C. Small nuclear RNAs from Saccharomyces cerevisiae: unexpected diversity in abundance, size, and molecular complexity. Proc Natl Acad Sci U S A. 1986 Nov;83(21):8097–8101. doi: 10.1073/pnas.83.21.8097. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Robinow C. F., Marak J. A fiber apparatus in the nucleus of the yeast cell. J Cell Biol. 1966 Apr;29(1):129–151. doi: 10.1083/jcb.29.1.129. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Schwarzacher H. G., Mikelsaar A. V., Schnedl W. The nature of the Ag-staining of nucleolus organizer regions. Electron- and light-microscopic studies on human cells in interphase, mitosis, and meiosis. Cytogenet Cell Genet. 1978;20(1-6):24–39. doi: 10.1159/000130837. [DOI] [PubMed] [Google Scholar]
  39. Sharp G. C., Irvin W. S., Tan E. M., Gould R. G., Holman H. R. Mixed connective tissue disease--an apparently distinct rheumatic disease syndrome associated with a specific antibody to an extractable nuclear antigen (ENA). Am J Med. 1972 Feb;52(2):148–159. doi: 10.1016/0002-9343(72)90064-2. [DOI] [PubMed] [Google Scholar]
  40. Smith H. C., Spector D. L., Woodcock C. L., Ochs R. L., Bhorjee J. Alterations in chromatin conformation are accompanied by reorganization of nonchromatin domains that contain U-snRNP protein p28 and nuclear protein p107. J Cell Biol. 1985 Aug;101(2):560–567. doi: 10.1083/jcb.101.2.560. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Smitt W. W., Vermeulen C. A., Vlak J. M., Rozijn T. H., Molenaar I. Electron microscopic autoradiographic study of RNA synthesis in yeast nucleus. Exp Cell Res. 1972 Jan;70(1):140–144. doi: 10.1016/0014-4827(72)90191-7. [DOI] [PubMed] [Google Scholar]
  42. Smitt W. W., Vlak J. M., Molenaar I., Rozijn T. H. Nucleolar function of the dense crescent in the yeast nucleus. A biochemical and ultrastructural study. Exp Cell Res. 1973 Aug;80(2):313–321. doi: 10.1016/0014-4827(73)90302-9. [DOI] [PubMed] [Google Scholar]
  43. Spector D. L. Colocalization of U1 and U2 small nuclear RNPs by immunocytochemistry. Biol Cell. 1984;51(1):109–112. doi: 10.1111/j.1768-322x.1984.tb00289.x. [DOI] [PubMed] [Google Scholar]
  44. Spector D. L. Higher order nuclear organization: three-dimensional distribution of small nuclear ribonucleoprotein particles. Proc Natl Acad Sci U S A. 1990 Jan;87(1):147–151. doi: 10.1073/pnas.87.1.147. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Spector D. L., Schrier W. H., Busch H. Immunoelectron microscopic localization of snRNPs. Biol Cell. 1983;49(1):1–10. doi: 10.1111/j.1768-322x.1984.tb00215.x. [DOI] [PubMed] [Google Scholar]
  46. Tanaka K., Kanbe T. Mitosis in the fission yeast Schizosaccharomyces pombe as revealed by freeze-substitution electron microscopy. J Cell Sci. 1986 Feb;80:253–268. doi: 10.1242/jcs.80.1.253. [DOI] [PubMed] [Google Scholar]
  47. Tyc K., Steitz J. A. U3, U8 and U13 comprise a new class of mammalian snRNPs localized in the cell nucleolus. EMBO J. 1989 Oct;8(10):3113–3119. doi: 10.1002/j.1460-2075.1989.tb08463.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Verheijen R., Kuijpers H., Vooijs P., Van Venrooij W., Ramaekers F. Distribution of the 70K U1 RNA-associated protein during interphase and mitosis. Correlation with other U RNP particles and proteins of the nuclear matrix. J Cell Sci. 1986 Dec;86:173–190. doi: 10.1242/jcs.86.1.173. [DOI] [PubMed] [Google Scholar]
  49. Williams M. A., Kleinschmidt J. A., Krohne G., Franke W. W. Argyrophilic nuclear and nucleolar proteins of Xenopus laevis oocytes identified by gel electrophoresis. Exp Cell Res. 1982 Feb;137(2):341–351. doi: 10.1016/0014-4827(82)90035-0. [DOI] [PubMed] [Google Scholar]
  50. Wise J. A., Tollervey D., Maloney D., Swerdlow H., Dunn E. J., Guthrie C. Yeast contains small nuclear RNAs encoded by single copy genes. Cell. 1983 Dec;35(3 Pt 2):743–751. doi: 10.1016/0092-8674(83)90107-1. [DOI] [PubMed] [Google Scholar]

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