Skip to main content
Biochemical Journal logoLink to Biochemical Journal
. 1999 Jan 1;337(Pt 1):29–35.

A conserved motif in the yeast nucleolar protein Nop2p contains an essential cysteine residue.

M King 1, D Ton 1, K L Redman 1
PMCID: PMC1219932  PMID: 9854021

Abstract

Nop2p is an essential nucleolar protein in Saccharomyces cerevisiae that is involved in large ribosomal subunit assembly. It has substantial homology with human p120, the proliferation-associated nucleolar antigen that is overexpressed in many human cancers. A motif containing an invariant Pro-Cys dipeptide is found in Nop2p, p120 and the bacterial Fmu proteins. A total of nine conserved residues, including Pro423 and Cys424, were individually altered in Nop2p by site-directed mutagenesis. Nop2p function was abolished by conversion of Cys424 into either alanine or serine. All of the other Nop2p mutations tested sustained yeast viability, including glycine replacement of Pro423 and the conversion of a second conserved cysteine into alanine. The crucial role of Cys424 in Nop2p is intriguing, due to the critical roles that cysteine residues adjacent to a proline have in a number of nucleotide-modifying enzymes.

Full Text

The Full Text of this article is available as a PDF (177.2 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Busch H. The final common pathway of cancer. Cancer Res. 1990 Aug 15;50(16):4830–4838. [PubMed] [Google Scholar]
  2. Busch R. K., Perlaky L., Valdez B. C., Henning D., Busch H. Apoptosis in human tumor cells following treatment with p120 antisense oligodeoxynucleotide ISIS 3466. Cancer Lett. 1994 Nov 11;86(2):151–157. doi: 10.1016/0304-3835(94)90073-6. [DOI] [PubMed] [Google Scholar]
  3. Chen L., MacMillan A. M., Chang W., Ezaz-Nikpay K., Lane W. S., Verdine G. L. Direct identification of the active-site nucleophile in a DNA (cytosine-5)-methyltransferase. Biochemistry. 1991 Nov 19;30(46):11018–11025. doi: 10.1021/bi00110a002. [DOI] [PubMed] [Google Scholar]
  4. Elble R. A simple and efficient procedure for transformation of yeasts. Biotechniques. 1992 Jul;13(1):18–20. [PubMed] [Google Scholar]
  5. Fonagy A., Swiderski C., Dunn M., Freeman J. W. Antisense-mediated specific inhibition of P120 protein expression prevents G1- to S-phase transition. Cancer Res. 1992 Oct 1;52(19):5250–5256. [PubMed] [Google Scholar]
  6. Fonagy A., Swiderski C., Ostrovsky A. M., Bolton W. E., Freeman J. W. Effect of nucleolar P120 expression level on the proliferation capacity of breast cancer cells. Cancer Res. 1994 Apr 1;54(7):1859–1864. [PubMed] [Google Scholar]
  7. Freeman J. W., Busch R. K., Gyorkey F., Gyorkey P., Ross B. E., Busch H. Identification and characterization of a human proliferation-associated nucleolar antigen with a molecular weight of 120,000 expressed in early G1 phase. Cancer Res. 1988 Mar 1;48(5):1244–1251. [PubMed] [Google Scholar]
  8. Freeman J. W., McGrath P., Bondada V., Selliah N., Ownby H., Maloney T., Busch R. K., Busch H. Prognostic significance of proliferation associated nucleolar antigen P120 in human breast carcinoma. Cancer Res. 1991 Apr 15;51(8):1973–1978. [PubMed] [Google Scholar]
  9. Hong B., Brockenbrough J. S., Wu P., Aris J. P. Nop2p is required for pre-rRNA processing and 60S ribosome subunit synthesis in yeast. Mol Cell Biol. 1997 Jan;17(1):378–388. doi: 10.1128/mcb.17.1.378. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Hough R. F., Bass B. L. Analysis of Xenopus dsRNA adenosine deaminase cDNAs reveals similarities to DNA methyltransferases. RNA. 1997 Apr;3(4):356–370. [PMC free article] [PubMed] [Google Scholar]
  11. Koonin E. V. Prediction of an rRNA methyltransferase domain in human tumor-specific nucleolar protein P120. Nucleic Acids Res. 1994 Jul 11;22(13):2476–2478. doi: 10.1093/nar/22.13.2476. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Kossykh V. G., Schlagman S. L., Hattman S. Studies on the function of conserved sequence motifs in the T4 Dam-[N6-adenine] and EcoRII [C5-cytosine] DNA methyltransferases. Gene. 1995 May 19;157(1-2):125–126. doi: 10.1016/0378-1119(94)00636-7. [DOI] [PubMed] [Google Scholar]
  13. Lai F., Drakas R., Nishikura K. Mutagenic analysis of double-stranded RNA adenosine deaminase, a candidate enzyme for RNA editing of glutamate-gated ion channel transcripts. J Biol Chem. 1995 Jul 21;270(29):17098–17105. doi: 10.1074/jbc.270.29.17098. [DOI] [PubMed] [Google Scholar]
  14. Malone T., Blumenthal R. M., Cheng X. Structure-guided analysis reveals nine sequence motifs conserved among DNA amino-methyltransferases, and suggests a catalytic mechanism for these enzymes. J Mol Biol. 1995 Nov 3;253(4):618–632. doi: 10.1006/jmbi.1995.0577. [DOI] [PubMed] [Google Scholar]
  15. McGrath P. C., Holley D. T., Hamby L. S., Mattingly C. A., Freeman J. W. Prospective study correlating P120 antigen expression with established prognostic factors in breast cancer. Surg Oncol. 1994 Apr;3(2):69–77. doi: 10.1016/0960-7404(94)90002-7. [DOI] [PubMed] [Google Scholar]
  16. McGrath P. C., Holley D. T., Hamby L. S., Powell D. E., Mattingly C., Freeman J. W. Proliferation-associated nucleolar antigen P120: a prognostic marker in node-negative breast cancer. Surgery. 1994 Oct;116(4):616–21. [PubMed] [Google Scholar]
  17. Navaratnam N., Bhattacharya S., Fujino T., Patel D., Jarmuz A. L., Scott J. Evolutionary origins of apoB mRNA editing: catalysis by a cytidine deaminase that has acquired a novel RNA-binding motif at its active site. Cell. 1995 Apr 21;81(2):187–195. doi: 10.1016/0092-8674(95)90328-3. [DOI] [PubMed] [Google Scholar]
  18. Pósfai J., Bhagwat A. S., Pósfai G., Roberts R. J. Predictive motifs derived from cytosine methyltransferases. Nucleic Acids Res. 1989 Apr 11;17(7):2421–2435. doi: 10.1093/nar/17.7.2421. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Redman K. L., Rechsteiner M. Extended reading frame of a ubiquitin gene encodes a stable, conserved, basic protein. J Biol Chem. 1988 Apr 5;263(10):4926–4931. [PubMed] [Google Scholar]
  20. Redman K. L. The smaller protein formed as a ubiquitin fusion in Drosophila is processed from ubiquitin and found on the 60S ribosomal subunit. Insect Biochem Mol Biol. 1994 Feb;24(2):191–201. doi: 10.1016/0965-1748(94)90085-x. [DOI] [PubMed] [Google Scholar]
  21. Ren Y., Busch R., Durban E., Taylor C., Gustafson W. C., Valdez B., Li Y. P., Smetana K., Busch H. Overexpression of human nucleolar proteins in insect cells: characterization of nucleolar protein p120. Protein Expr Purif. 1996 Mar;7(2):212–219. doi: 10.1006/prep.1996.0029. [DOI] [PubMed] [Google Scholar]
  22. Santi D. V., McHenry C. S., Raines R. T., Ivanetich K. M. Kinetics and thermodynamics of the interaction of 5-fluoro-2'-deoxyuridylate with thymidylate synthase. Biochemistry. 1987 Dec 29;26(26):8606–8613. doi: 10.1021/bi00400a017. [DOI] [PubMed] [Google Scholar]
  23. Schuler G. D., Altschul S. F., Lipman D. J. A workbench for multiple alignment construction and analysis. Proteins. 1991;9(3):180–190. doi: 10.1002/prot.340090304. [DOI] [PubMed] [Google Scholar]
  24. 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]
  25. Sikorski R. S., Hieter P. A system of shuttle vectors and yeast host strains designed for efficient manipulation of DNA in Saccharomyces cerevisiae. Genetics. 1989 May;122(1):19–27. doi: 10.1093/genetics/122.1.19. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Ueki T., Nakayama Y., Sugao Y., Kohno K., Matsuo K., Sugimoto Y., Yamada Y., Kuwano M., Tsuneyoshi M. Significance of the expression of proliferation-associated nucleolar antigen p120 in human colorectal tumors. Hum Pathol. 1997 Jan;28(1):74–79. doi: 10.1016/s0046-8177(97)90282-3. [DOI] [PubMed] [Google Scholar]
  27. Valdez B. C., Perlaky L., Saijo Y., Henning D., Zhu C., Busch R. K., Zhang W. W., Busch H. A region of antisense RNA from human p120 cDNA with high homology to mouse p120 cDNA inhibits NIH 3T3 proliferation. Cancer Res. 1992 Oct 15;52(20):5681–5686. [PubMed] [Google Scholar]
  28. de Beus E., Brockenbrough J. S., Hong B., Aris J. P. Yeast NOP2 encodes an essential nucleolar protein with homology to a human proliferation marker. J Cell Biol. 1994 Dec;127(6 Pt 2):1799–1813. doi: 10.1083/jcb.127.6.1799. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

RESOURCES