Abstract
Using monoclonal antibodies as probes, we have characterized three antigens with respect to localization in the nucleolus, molecular weight and solubility. Two proteins, of 110,000 and 94,000 apparent molecular weight, were found associated with the ribonucleoprotein fibers. A third protein, with a molecular weight of 40,000, was accumulated at the nucleolar periphery, was present in the nucleoplasm, and may be involved in pre-ribosome maturation and transport.
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- Attardi G., Amaldi F. Structure and synthesis of ribosomal RNA. Annu Rev Biochem. 1970;39:183–226. doi: 10.1146/annurev.bi.39.070170.001151. [DOI] [PubMed] [Google Scholar]
- Blumenthal A. B., Dieden J. D., Kapp L. N., Sedat J. W. Rapid isolation of metaphase chromosomes containing high molecular weight DNA. J Cell Biol. 1979 Apr;81(1):255–259. doi: 10.1083/jcb.81.1.255. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bowen B., Steinberg J., Laemmli U. K., Weintraub H. The detection of DNA-binding proteins by protein blotting. Nucleic Acids Res. 1980 Jan 11;8(1):1–20. doi: 10.1093/nar/8.1.1. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- Johnson G. D., Nogueira Araujo G. M. A simple method of reducing the fading of immunofluorescence during microscopy. J Immunol Methods. 1981;43(3):349–350. doi: 10.1016/0022-1759(81)90183-6. [DOI] [PubMed] [Google Scholar]
- Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
- Laskey R. A., Mills A. D. Enhanced autoradiographic detection of 32P and 125I using intensifying screens and hypersensitized film. FEBS Lett. 1977 Oct 15;82(2):314–316. doi: 10.1016/0014-5793(77)80609-1. [DOI] [PubMed] [Google Scholar]
- Lebkowski J. S., Laemmli U. K. Evidence for two levels of DNA folding in histone-depleted HeLa interphase nuclei. J Mol Biol. 1982 Apr 5;156(2):309–324. doi: 10.1016/0022-2836(82)90331-x. [DOI] [PubMed] [Google Scholar]
- Lebkowski J. S., Laemmli U. K. Non-histone proteins and long-range organization of HeLa interphase DNA. J Mol Biol. 1982 Apr 5;156(2):325–344. doi: 10.1016/0022-2836(82)90332-1. [DOI] [PubMed] [Google Scholar]
- Lewis C. D., Laemmli U. K. Higher order metaphase chromosome structure: evidence for metalloprotein interactions. Cell. 1982 May;29(1):171–181. doi: 10.1016/0092-8674(82)90101-5. [DOI] [PubMed] [Google Scholar]
- Lischwe M. A., Richards R. L., Busch R. K., Busch H. Localization of phosphoprotein C23 to nucleolar structures and to the nucleolus organizer regions. Exp Cell Res. 1981 Nov;136(1):101–109. doi: 10.1016/0014-4827(81)90041-0. [DOI] [PubMed] [Google Scholar]
- Mamrack M. D., Olson M. O., Busch H. Amino acid sequence and sites of phosphorylation in a highly acidic region of nucleolar nonhistone protein C23. Biochemistry. 1979 Jul 24;18(15):3381–3386. doi: 10.1021/bi00582a026. [DOI] [PubMed] [Google Scholar]
- Michalik J., Yeoman L. C., Busch H. Nucleolar localization of protein B23 (37/5.1) by immunocytochemical techniques. Life Sci. 1981 Mar 23;28(12):1371–1379. doi: 10.1016/0024-3205(81)90411-2. [DOI] [PubMed] [Google Scholar]
- Miller O. L., Jr The nucleolus, chromosomes, and visualization of genetic activity. J Cell Biol. 1981 Dec;91(3 Pt 2):15s–27s. doi: 10.1083/jcb.91.3.15s. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Olson M. O., Busch H. Nucleolar proteins. Methods Cell Biol. 1978;17:163–210. [PubMed] [Google Scholar]
- Olson M. O., Ezrailson E. G., Guetzow K., Busch H. Localization and phosphorylation of nuclear, nucleolar and extranucleolar non-histone proteins of Novikoff hepatoma ascites cells. J Mol Biol. 1975 Oct 5;97(4):611–619. doi: 10.1016/s0022-2836(75)80062-3. [DOI] [PubMed] [Google Scholar]
- Olson M. O., Guetzow K., Busch H. Localization of phosphoprotein C23 in nucleoli by immunological methods. Exp Cell Res. 1981 Oct;135(2):259–265. doi: 10.1016/0014-4827(81)90161-0. [DOI] [PubMed] [Google Scholar]
- Olson M. O., Rivers Z. M., Thompson B. A., Kao W. Y., Case S. T. Interaction of nucleolar phosphoprotein C23 with cloned segments of rat ribosomal deoxyribonucleic acid. Biochemistry. 1983 Jul 5;22(14):3345–3351. doi: 10.1021/bi00283a007. [DOI] [PubMed] [Google Scholar]
- Olson M. O., Thompson B. A. Distribution of proteins among chromatin components of nucleoli. Biochemistry. 1983 Jun 21;22(13):3187–3193. doi: 10.1021/bi00282a023. [DOI] [PubMed] [Google Scholar]
- Orrick L. R., Olson M. O., Busch H. Comparison of nucleolar proteins of normal rat liver and Novikoff hepatoma ascites cells by two-dimensional polyacrylamide gel electrophoresis. Proc Natl Acad Sci U S A. 1973 May;70(5):1316–1320. doi: 10.1073/pnas.70.5.1316. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Renart J., Reiser J., Stark G. R. Transfer of proteins from gels to diazobenzyloxymethyl-paper and detection with antisera: a method for studying antibody specificity and antigen structure. Proc Natl Acad Sci U S A. 1979 Jul;76(7):3116–3120. doi: 10.1073/pnas.76.7.3116. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Rothblum L. I., Mamrack P. M., Kunkle H. M., Olson M. O., Busch H. Fractionation of nucleoli. Enzymatic and two-dimensional polyacrylamide gel electrophoretic analysis. Biochemistry. 1977 Oct 18;16(21):4716–4721. doi: 10.1021/bi00640a028. [DOI] [PubMed] [Google Scholar]
- Stähli C., Staehelin T., Miggiano V., Schmidt J., Häring P. High frequencies of antigen-specific hybridomas: dependence on immunization parameters and prediction by spleen cell analysis. J Immunol Methods. 1980;32(3):297–304. doi: 10.1016/0022-1759(80)90194-5. [DOI] [PubMed] [Google Scholar]
- Stähli C., Staehelin T., Miggiano V. Spleen cell analysis and optimal immunization for high-frequency production of specific hybridomas. Methods Enzymol. 1983;92:26–36. doi: 10.1016/0076-6879(83)92006-2. [DOI] [PubMed] [Google Scholar]
- Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Tsutsui K., Tsutsui K., Oda T. Isolation and characterization of a high-molecular-weight acid-soluble nuclear protein from mouse ascites-sarcoma cells. Eur J Biochem. 1980 Jul;108(2):497–505. doi: 10.1111/j.1432-1033.1980.tb04747.x. [DOI] [PubMed] [Google Scholar]