Skip to main content
NCBI home page
Nucleic Acids Research logoLink to Nucleic Acids Research
. 1998 Sep 1;26(17):3908–3914. doi: 10.1093/nar/26.17.3908

The products of the yeast MMS2 and two human homologs (hMMS2 and CROC-1) define a structurally and functionally conserved Ubc-like protein family.

W Xiao 1, S L Lin 1, S Broomfield 1, B L Chow 1, Y F Wei 1
PMCID: PMC147796  PMID: 9705497

Abstract

Eukaryotic genes encoding ubiquitin-congugating enzyme (Ubc)-like proteins have been isolated from both human and yeast cells. The CROC-1 gene was isolated by its ability to transactivate c- fos expression in cell culture through a tandem repeat enhancer sequence. The yeast MMS2 gene was cloned by its ability to complement the methyl methanesulfonate sensitivity of the mms2-1 mutant and was later shown to be involved in DNA post-replication repair. We report here the identification of a human MMS2 ( hMMS2 ) cDNA encoding a novel human Ubc-like protein. hMMS2 and CROC-1 share >90% amino acid sequence identity, but their DNA probes hybridize to distinct transcripts. hMMS2 and CROC-1 also share approximately 50% identity and 75% similarity with the entire length of yeast Mms2. Unlike CROC-1 , whose transcript appears to be elevated in all tumor cell lines examined, the hMMS2 transcript is only elevated in some tumor cell lines. Collectively, these results indicate that eukaryotic cells may contain a highly conserved family of Ubc-like proteins that play roles in diverse cellular processes, ranging from DNA repair to signal transduction and cell differentiation. The hMMS2 and CROC-1 genes are able to functionally complement the yeast mms2 defects with regard to sensitivity to DNA damaging agents and spontaneous mutagenesis. Conversely, both MMS2 and hMMS2 were able to transactivate a c- fos - CAT reporter gene in Rat-1 cells in a transient co-transfection assay. We propose that either these proteins function in a common cellular process, such as DNA repair, or they exert their diverse biological roles through a similar biochemical interaction relative to ubiquitination.

Full Text

The Full Text of this article is available as a PDF (238.5 KB).

Selected References

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

  1. Bailly V., Lamb J., Sung P., Prakash S., Prakash L. Specific complex formation between yeast RAD6 and RAD18 proteins: a potential mechanism for targeting RAD6 ubiquitin-conjugating activity to DNA damage sites. Genes Dev. 1994 Apr 1;8(7):811–820. doi: 10.1101/gad.8.7.811. [DOI] [PubMed] [Google Scholar]
  2. Banerjee A., Deshaies R. J., Chau V. Characterization of a dominant negative mutant of the cell cycle ubiquitin-conjugating enzyme Cdc34. J Biol Chem. 1995 Nov 3;270(44):26209–26215. doi: 10.1074/jbc.270.44.26209. [DOI] [PubMed] [Google Scholar]
  3. Bartel B., Wünning I., Varshavsky A. The recognition component of the N-end rule pathway. EMBO J. 1990 Oct;9(10):3179–3189. doi: 10.1002/j.1460-2075.1990.tb07516.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Brinkmann U., Gallo M., Polymeropoulos M. H., Pastan I. The human CAS (cellular apoptosis susceptibility) gene mapping on chromosome 20q13 is amplified in BT474 breast cancer cells and part of aberrant chromosomes in breast and colon cancer cell lines. Genome Res. 1996 Mar;6(3):187–194. doi: 10.1101/gr.6.3.187. [DOI] [PubMed] [Google Scholar]
  5. Broomfield S., Chow B. L., Xiao W. MMS2, encoding a ubiquitin-conjugating-enzyme-like protein, is a member of the yeast error-free postreplication repair pathway. Proc Natl Acad Sci U S A. 1998 May 12;95(10):5678–5683. doi: 10.1073/pnas.95.10.5678. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Chen P., Johnson P., Sommer T., Jentsch S., Hochstrasser M. Multiple ubiquitin-conjugating enzymes participate in the in vivo degradation of the yeast MAT alpha 2 repressor. Cell. 1993 Jul 30;74(2):357–369. doi: 10.1016/0092-8674(93)90426-q. [DOI] [PubMed] [Google Scholar]
  7. Cohen-Jonathan E., Toulas C., Monteil S., Couderc B., Maret A., Bard J. J., Prats H., Daly-Schveitzer N., Favre G. Radioresistance induced by the high molecular forms of the basic fibroblast growth factor is associated with an increased G2 delay and a hyperphosphorylation of p34CDC2 in HeLa cells. Cancer Res. 1997 Apr 1;57(7):1364–1370. [PubMed] [Google Scholar]
  8. Cook W. J., Jeffrey L. C., Sullivan M. L., Vierstra R. D. Three-dimensional structure of a ubiquitin-conjugating enzyme (E2). J Biol Chem. 1992 Jul 25;267(21):15116–15121. doi: 10.2210/pdb1aak/pdb. [DOI] [PubMed] [Google Scholar]
  9. Cook W. J., Jeffrey L. C., Xu Y., Chau V. Tertiary structures of class I ubiquitin-conjugating enzymes are highly conserved: crystal structure of yeast Ubc4. Biochemistry. 1993 Dec 21;32(50):13809–13817. doi: 10.1021/bi00213a009. [DOI] [PubMed] [Google Scholar]
  10. Cook W. J., Martin P. D., Edwards B. F., Yamazaki R. K., Chau V. Crystal structure of a class I ubiquitin conjugating enzyme (Ubc7) from Saccharomyces cerevisiae at 2.9 angstroms resolution. Biochemistry. 1997 Feb 18;36(7):1621–1627. doi: 10.1021/bi962639e. [DOI] [PubMed] [Google Scholar]
  11. Dohmen R. J., Madura K., Bartel B., Varshavsky A. The N-end rule is mediated by the UBC2(RAD6) ubiquitin-conjugating enzyme. Proc Natl Acad Sci U S A. 1991 Aug 15;88(16):7351–7355. doi: 10.1073/pnas.88.16.7351. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Edelman A. M., Blumenthal D. K., Krebs E. G. Protein serine/threonine kinases. Annu Rev Biochem. 1987;56:567–613. doi: 10.1146/annurev.bi.56.070187.003031. [DOI] [PubMed] [Google Scholar]
  13. El-Rifai W., Harper J. C., Cummings O. W., Hyytinen E. R., Frierson H. F., Jr, Knuutila S., Powell S. M. Consistent genetic alterations in xenografts of proximal stomach and gastro-esophageal junction adenocarcinomas. Cancer Res. 1998 Jan 1;58(1):34–37. [PubMed] [Google Scholar]
  14. Feldman R. M., Correll C. C., Kaplan K. B., Deshaies R. J. A complex of Cdc4p, Skp1p, and Cdc53p/cullin catalyzes ubiquitination of the phosphorylated CDK inhibitor Sic1p. Cell. 1997 Oct 17;91(2):221–230. doi: 10.1016/s0092-8674(00)80404-3. [DOI] [PubMed] [Google Scholar]
  15. Finley D., Bartel B., Varshavsky A. The tails of ubiquitin precursors are ribosomal proteins whose fusion to ubiquitin facilitates ribosome biogenesis. Nature. 1989 Mar 30;338(6214):394–401. doi: 10.1038/338394a0. [DOI] [PubMed] [Google Scholar]
  16. Finley D., Chau V. Ubiquitination. Annu Rev Cell Biol. 1991;7:25–69. doi: 10.1146/annurev.cb.07.110191.000325. [DOI] [PubMed] [Google Scholar]
  17. Finley D., Ozkaynak E., Varshavsky A. The yeast polyubiquitin gene is essential for resistance to high temperatures, starvation, and other stresses. Cell. 1987 Mar 27;48(6):1035–1046. doi: 10.1016/0092-8674(87)90711-2. [DOI] [PubMed] [Google Scholar]
  18. Fritsche J., Rehli M., Krause S. W., Andreesen R., Kreutz M. Molecular cloning of a 1alpha,25-dihydroxyvitamin D3-inducible transcript (DDVit 1) in human blood monocytes. Biochem Biophys Res Commun. 1997 Jun 18;235(2):407–412. doi: 10.1006/bbrc.1997.6798. [DOI] [PubMed] [Google Scholar]
  19. Glass D. B., el-Maghrabi M. R., Pilkis S. J. Synthetic peptides corresponding to the site phosphorylated in 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase as substrates of cyclic nucleotide-dependent protein kinases. J Biol Chem. 1986 Feb 25;261(6):2987–2993. [PubMed] [Google Scholar]
  20. Goebl M. G., Yochem J., Jentsch S., McGrath J. P., Varshavsky A., Byers B. The yeast cell cycle gene CDC34 encodes a ubiquitin-conjugating enzyme. Science. 1988 Sep 9;241(4871):1331–1335. doi: 10.1126/science.2842867. [DOI] [PubMed] [Google Scholar]
  21. Gutman A., Wasylyk C., Wasylyk B. Cell-specific regulation of oncogene-responsive sequences of the c-fos promoter. Mol Cell Biol. 1991 Oct;11(10):5381–5387. doi: 10.1128/mcb.11.10.5381. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Hochstrasser M. There's the rub: a novel ubiquitin-like modification linked to cell cycle regulation. Genes Dev. 1998 Apr 1;12(7):901–907. doi: 10.1101/gad.12.7.901. [DOI] [PubMed] [Google Scholar]
  23. Jentsch S., McGrath J. P., Varshavsky A. The yeast DNA repair gene RAD6 encodes a ubiquitin-conjugating enzyme. Nature. 1987 Sep 10;329(6135):131–134. doi: 10.1038/329131a0. [DOI] [PubMed] [Google Scholar]
  24. Jentsch S. The ubiquitin-conjugation system. Annu Rev Genet. 1992;26:179–207. doi: 10.1146/annurev.ge.26.120192.001143. [DOI] [PubMed] [Google Scholar]
  25. Johnson E. S., Blobel G. Ubc9p is the conjugating enzyme for the ubiquitin-like protein Smt3p. J Biol Chem. 1997 Oct 24;272(43):26799–26802. doi: 10.1074/jbc.272.43.26799. [DOI] [PubMed] [Google Scholar]
  26. Kaina B., Haas S., Kappes H. A general role for c-Fos in cellular protection against DNA-damaging carcinogens and cytostatic drugs. Cancer Res. 1997 Jul 1;57(13):2721–2731. [PubMed] [Google Scholar]
  27. Kallioniemi A., Kallioniemi O. P., Piper J., Tanner M., Stokke T., Chen L., Smith H. S., Pinkel D., Gray J. W., Waldman F. M. Detection and mapping of amplified DNA sequences in breast cancer by comparative genomic hybridization. Proc Natl Acad Sci U S A. 1994 Mar 15;91(6):2156–2160. doi: 10.1073/pnas.91.6.2156. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Kishimoto A., Nishiyama K., Nakanishi H., Uratsuji Y., Nomura H., Takeyama Y., Nishizuka Y. Studies on the phosphorylation of myelin basic protein by protein kinase C and adenosine 3':5'-monophosphate-dependent protein kinase. J Biol Chem. 1985 Oct 15;260(23):12492–12499. [PubMed] [Google Scholar]
  29. Koonin E. V., Abagyan R. A. TSG101 may be the prototype of a class of dominant negative ubiquitin regulators. Nat Genet. 1997 Aug;16(4):330–331. doi: 10.1038/ng0897-330. [DOI] [PubMed] [Google Scholar]
  30. Koonin E. V., Bork P., Sander C. Yeast chromosome III: new gene functions. EMBO J. 1994 Feb 1;13(3):493–503. doi: 10.1002/j.1460-2075.1994.tb06287.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Lammer D., Mathias N., Laplaza J. M., Jiang W., Liu Y., Callis J., Goebl M., Estelle M. Modification of yeast Cdc53p by the ubiquitin-related protein rub1p affects function of the SCFCdc4 complex. Genes Dev. 1998 Apr 1;12(7):914–926. doi: 10.1101/gad.12.7.914. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Lee M. P., Feinberg A. P. Aberrant splicing but not mutations of TSG101 in human breast cancer. Cancer Res. 1997 Aug 1;57(15):3131–3134. [PubMed] [Google Scholar]
  33. Li L., Cohen S. N. Tsg101: a novel tumor susceptibility gene isolated by controlled homozygous functional knockout of allelic loci in mammalian cells. Cell. 1996 May 3;85(3):319–329. doi: 10.1016/s0092-8674(00)81111-3. [DOI] [PubMed] [Google Scholar]
  34. Li L., Li X., Francke U., Cohen S. N. The TSG101 tumor susceptibility gene is located in chromosome 11 band p15 and is mutated in human breast cancer. Cell. 1997 Jan 10;88(1):143–154. doi: 10.1016/s0092-8674(00)81866-8. [DOI] [PubMed] [Google Scholar]
  35. Liakopoulos D., Doenges G., Matuschewski K., Jentsch S. A novel protein modification pathway related to the ubiquitin system. EMBO J. 1998 Apr 15;17(8):2208–2214. doi: 10.1093/emboj/17.8.2208. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Lisztwan J., Marti A., Sutterlüty H., Gstaiger M., Wirbelauer C., Krek W. Association of human CUL-1 and ubiquitin-conjugating enzyme CDC34 with the F-box protein p45(SKP2): evidence for evolutionary conservation in the subunit composition of the CDC34-SCF pathway. EMBO J. 1998 Jan 15;17(2):368–383. doi: 10.1093/emboj/17.2.368. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Müller S., Matunis M. J., Dejean A. Conjugation with the ubiquitin-related modifier SUMO-1 regulates the partitioning of PML within the nucleus. EMBO J. 1998 Jan 2;17(1):61–70. doi: 10.1093/emboj/17.1.61. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Peng C. Y., Graves P. R., Thoma R. S., Wu Z., Shaw A. S., Piwnica-Worms H. Mitotic and G2 checkpoint control: regulation of 14-3-3 protein binding by phosphorylation of Cdc25C on serine-216. Science. 1997 Sep 5;277(5331):1501–1505. doi: 10.1126/science.277.5331.1501. [DOI] [PubMed] [Google Scholar]
  39. Plon S. E., Leppig K. A., Do H. N., Groudine M. Cloning of the human homolog of the CDC34 cell cycle gene by complementation in yeast. Proc Natl Acad Sci U S A. 1993 Nov 15;90(22):10484–10488. doi: 10.1073/pnas.90.22.10484. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Ponting C. P., Cai Y. D., Bork P. The breast cancer gene product TSG101: a regulator of ubiquitination? J Mol Med (Berl) 1997 Jul;75(7):467–469. [PubMed] [Google Scholar]
  41. Prakash L., Prakash S. Isolation and characterization of MMS-sensitive mutants of Saccharomyces cerevisiae. Genetics. 1977 May;86(1):33–55. doi: 10.1093/genetics/86.1.33. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Ptak C., Prendergast J. A., Hodgins R., Kay C. M., Chau V., Ellison M. J. Functional and physical characterization of the cell cycle ubiquitin-conjugating enzyme CDC34 (UBC3). Identification of a functional determinant within the tail that facilitates CDC34 self-association. J Biol Chem. 1994 Oct 21;269(42):26539–26545. [PubMed] [Google Scholar]
  43. Rothofsky M. L., Lin S. L. CROC-1 encodes a protein which mediates transcriptional activation of the human FOS promoter. Gene. 1997 Aug 22;195(2):141–149. doi: 10.1016/s0378-1119(97)00097-8. [DOI] [PubMed] [Google Scholar]
  44. Rothstein R. J. One-step gene disruption in yeast. Methods Enzymol. 1983;101:202–211. doi: 10.1016/0076-6879(83)01015-0. [DOI] [PubMed] [Google Scholar]
  45. Saitoh H., Pu R. T., Dasso M. SUMO-1: wrestling with a new ubiquitin-related modifier. Trends Biochem Sci. 1997 Oct;22(10):374–376. doi: 10.1016/s0968-0004(97)01102-x. [DOI] [PubMed] [Google Scholar]
  46. Sanchez Y., Wong C., Thoma R. S., Richman R., Wu Z., Piwnica-Worms H., Elledge S. J. Conservation of the Chk1 checkpoint pathway in mammals: linkage of DNA damage to Cdk regulation through Cdc25. Science. 1997 Sep 5;277(5331):1497–1501. doi: 10.1126/science.277.5331.1497. [DOI] [PubMed] [Google Scholar]
  47. Sancho E., Vilá M. R., Sánchez-Pulido L., Lozano J. J., Paciucci R., Nadal M., Fox M., Harvey C., Bercovich B., Loukili N. Role of UEV-1, an inactive variant of the E2 ubiquitin-conjugating enzymes, in in vitro differentiation and cell cycle behavior of HT-29-M6 intestinal mucosecretory cells. Mol Cell Biol. 1998 Jan;18(1):576–589. doi: 10.1128/mcb.18.1.576. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  49. Savelieva E., Belair C. D., Newton M. A., DeVries S., Gray J. W., Waldman F., Reznikoff C. A. 20q gain associates with immortalization: 20q13.2 amplification correlates with genome instability in human papillomavirus 16 E7 transformed human uroepithelial cells. Oncogene. 1997 Feb 6;14(5):551–560. doi: 10.1038/sj.onc.1200868. [DOI] [PubMed] [Google Scholar]
  50. Schwab M., Lutum A. S., Seufert W. Yeast Hct1 is a regulator of Clb2 cyclin proteolysis. Cell. 1997 Aug 22;90(4):683–693. doi: 10.1016/s0092-8674(00)80529-2. [DOI] [PubMed] [Google Scholar]
  51. Schwarz S. E., Matuschewski K., Liakopoulos D., Scheffner M., Jentsch S. The ubiquitin-like proteins SMT3 and SUMO-1 are conjugated by the UBC9 E2 enzyme. Proc Natl Acad Sci U S A. 1998 Jan 20;95(2):560–564. doi: 10.1073/pnas.95.2.560. [DOI] [PMC free article] [PubMed] [Google Scholar]
  52. Seufert W., Jentsch S. Ubiquitin-conjugating enzymes UBC4 and UBC5 mediate selective degradation of short-lived and abnormal proteins. EMBO J. 1990 Feb;9(2):543–550. doi: 10.1002/j.1460-2075.1990.tb08141.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  53. Skowyra D., Craig K. L., Tyers M., Elledge S. J., Harper J. W. F-box proteins are receptors that recruit phosphorylated substrates to the SCF ubiquitin-ligase complex. Cell. 1997 Oct 17;91(2):209–219. doi: 10.1016/s0092-8674(00)80403-1. [DOI] [PubMed] [Google Scholar]
  54. Sung P., Berleth E., Pickart C., Prakash S., Prakash L. Yeast RAD6 encoded ubiquitin conjugating enzyme mediates protein degradation dependent on the N-end-recognizing E3 enzyme. EMBO J. 1991 Aug;10(8):2187–2193. doi: 10.1002/j.1460-2075.1991.tb07754.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  55. Sung P., Prakash S., Prakash L. Mutation of cysteine-88 in the Saccharomyces cerevisiae RAD6 protein abolishes its ubiquitin-conjugating activity and its various biological functions. Proc Natl Acad Sci U S A. 1990 Apr;87(7):2695–2699. doi: 10.1073/pnas.87.7.2695. [DOI] [PMC free article] [PubMed] [Google Scholar]
  56. Tanner M. M., Tirkkonen M., Kallioniemi A., Collins C., Stokke T., Karhu R., Kowbel D., Shadravan F., Hintz M., Kuo W. L. Increased copy number at 20q13 in breast cancer: defining the critical region and exclusion of candidate genes. Cancer Res. 1994 Aug 15;54(16):4257–4260. [PubMed] [Google Scholar]
  57. Tanner M. M., Tirkkonen M., Kallioniemi A., Holli K., Collins C., Kowbel D., Gray J. W., Kallioniemi O. P., Isola J. Amplification of chromosomal region 20q13 in invasive breast cancer: prognostic implications. Clin Cancer Res. 1995 Dec;1(12):1455–1461. [PubMed] [Google Scholar]
  58. Varshavsky A. The N-end rule: functions, mysteries, uses. Proc Natl Acad Sci U S A. 1996 Oct 29;93(22):12142–12149. doi: 10.1073/pnas.93.22.12142. [DOI] [PMC free article] [PubMed] [Google Scholar]
  59. Varshavsky A. The ubiquitin system. Trends Biochem Sci. 1997 Oct;22(10):383–387. doi: 10.1016/s0968-0004(97)01122-5. [DOI] [PubMed] [Google Scholar]
  60. Visintin R., Prinz S., Amon A. CDC20 and CDH1: a family of substrate-specific activators of APC-dependent proteolysis. Science. 1997 Oct 17;278(5337):460–463. doi: 10.1126/science.278.5337.460. [DOI] [PubMed] [Google Scholar]
  61. Von Borstel R. C. Measuring spontaneous mutation rates in yeast. Methods Cell Biol. 1978;20:1–24. doi: 10.1016/s0091-679x(08)62005-1. [DOI] [PubMed] [Google Scholar]
  62. Watkins J. F., Sung P., Prakash S., Prakash L. The extremely conserved amino terminus of RAD6 ubiquitin-conjugating enzyme is essential for amino-end rule-dependent protein degradation. Genes Dev. 1993 Feb;7(2):250–261. doi: 10.1101/gad.7.2.250. [DOI] [PubMed] [Google Scholar]
  63. Williamson M. S., Game J. C., Fogel S. Meiotic gene conversion mutants in Saccharomyces cerevisiae. I. Isolation and characterization of pms1-1 and pms1-2. Genetics. 1985 Aug;110(4):609–646. doi: 10.1093/genetics/110.4.609. [DOI] [PMC free article] [PubMed] [Google Scholar]
  64. Xiao W., Chow B. L., Rathgeber L. The repair of DNA methylation damage in Saccharomyces cerevisiae. Curr Genet. 1996 Dec;30(6):461–468. doi: 10.1007/s002940050157. [DOI] [PubMed] [Google Scholar]
  65. Yeager T. R., DeVries S., Jarrard D. F., Kao C., Nakada S. Y., Moon T. D., Bruskewitz R., Stadler W. M., Meisner L. F., Gilchrist K. W. Overcoming cellular senescence in human cancer pathogenesis. Genes Dev. 1998 Jan 15;12(2):163–174. doi: 10.1101/gad.12.2.163. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Nucleic Acids Research are provided here courtesy of Oxford University Press

RESOURCES