Abstract
The ubiquitin-activating enzyme E1 catalyzes the first step in ubiquitin conjugation. We have cloned and sequenced the cDNA for human E1. This clone predicts a protein of 110,450 Da. Cys-194 lies within a region of identity to active-site Cys-88 of the ubiquitin carrier protein E2, suggesting a potential role for this region in enzymatic function of this protein. In addition, Cys-454 lies within a region of identity to the thiol ester consensus sequence of several proteins involved in thioester formation. Tissue distribution reveals a single 3.5-kilobase E1 message ubiquitous among tissues and cell lines.
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Selected References
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- Aebersold R. H., Leavitt J., Saavedra R. A., Hood L. E., Kent S. B. Internal amino acid sequence analysis of proteins separated by one- or two-dimensional gel electrophoresis after in situ protease digestion on nitrocellulose. Proc Natl Acad Sci U S A. 1987 Oct;84(20):6970–6974. doi: 10.1073/pnas.84.20.6970. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ball E., Karlik C. C., Beall C. J., Saville D. L., Sparrow J. C., Bullard B., Fyrberg E. A. Arthrin, a myofibrillar protein of insect flight muscle, is an actin-ubiquitin conjugate. Cell. 1987 Oct 23;51(2):221–228. doi: 10.1016/0092-8674(87)90149-8. [DOI] [PubMed] [Google Scholar]
- Ciechanover A., Elias S., Heller H., Hershko A. "Covalent affinity" purification of ubiquitin-activating enzyme. J Biol Chem. 1982 Mar 10;257(5):2537–2542. [PubMed] [Google Scholar]
- Ciechanover A., Finley D., Varshavsky A. Ubiquitin dependence of selective protein degradation demonstrated in the mammalian cell cycle mutant ts85. Cell. 1984 May;37(1):57–66. doi: 10.1016/0092-8674(84)90300-3. [DOI] [PubMed] [Google Scholar]
- Cornish-Bowden A. Critical values for testing the significance of amino acid composition indexes. Anal Biochem. 1980 Jul 1;105(2):233–238. doi: 10.1016/0003-2697(80)90450-9. [DOI] [PubMed] [Google Scholar]
- Feinberg A. P., Vogelstein B. A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem. 1983 Jul 1;132(1):6–13. doi: 10.1016/0003-2697(83)90418-9. [DOI] [PubMed] [Google Scholar]
- Finley D., Ciechanover A., Varshavsky A. Thermolability of ubiquitin-activating enzyme from the mammalian cell cycle mutant ts85. Cell. 1984 May;37(1):43–55. doi: 10.1016/0092-8674(84)90299-x. [DOI] [PubMed] [Google Scholar]
- Fleming R. E., Gitlin J. D. Primary structure of rat ceruloplasmin and analysis of tissue-specific gene expression during development. J Biol Chem. 1990 May 5;265(13):7701–7707. [PubMed] [Google Scholar]
- Haas A. L., Rose I. A. The mechanism of ubiquitin activating enzyme. A kinetic and equilibrium analysis. J Biol Chem. 1982 Sep 10;257(17):10329–10337. [PubMed] [Google Scholar]
- Hatfield P. M., Callis J., Vierstra R. D. Cloning of ubiquitin activating enzyme from wheat and expression of a functional protein in Escherichia coli. J Biol Chem. 1990 Sep 15;265(26):15813–15817. [PubMed] [Google Scholar]
- Hostetter M. K., Gordon D. L. Biochemistry of C3 and related thiolester proteins in infection and inflammation. Rev Infect Dis. 1987 Jan-Feb;9(1):97–109. doi: 10.1093/clinids/9.1.97. [DOI] [PubMed] [Google Scholar]
- Hunkapiller M. W., Hewick R. M., Dreyer W. J., Hood L. E. High-sensitivity sequencing with a gas-phase sequenator. Methods Enzymol. 1983;91:399–413. doi: 10.1016/s0076-6879(83)91038-8. [DOI] [PubMed] [Google Scholar]
- Hunt L. T., Dayhoff M. O. Amino-terminal sequence identity of ubiquitin and the nonhistone component of nuclear protein A24. Biochem Biophys Res Commun. 1977 Jan 24;74(2):650–655. doi: 10.1016/0006-291x(77)90352-7. [DOI] [PubMed] [Google Scholar]
- Kulka R. G., Raboy B., Schuster R., Parag H. A., Diamond G., Ciechanover A., Marcus M. A Chinese hamster cell cycle mutant arrested at G2 phase has a temperature-sensitive ubiquitin-activating enzyme, E1. J Biol Chem. 1988 Oct 25;263(30):15726–15731. [PubMed] [Google Scholar]
- MacDonald R. J., Swift G. H., Przybyla A. E., Chirgwin J. M. Isolation of RNA using guanidinium salts. Methods Enzymol. 1987;152:219–227. doi: 10.1016/0076-6879(87)52023-7. [DOI] [PubMed] [Google Scholar]
- Mayer A., Gropper R., Schwartz A. L., Ciechanover A. Purification, characterization, and rapid inactivation of thermolabile ubiquitin-activating enzyme from the mammalian cell cycle mutant ts85. J Biol Chem. 1989 Feb 5;264(4):2060–2068. [PubMed] [Google Scholar]
- Miller H. Practical aspects of preparing phage and plasmid DNA: growth, maintenance, and storage of bacteria and bacteriophage. Methods Enzymol. 1987;152:145–170. doi: 10.1016/0076-6879(87)52016-x. [DOI] [PubMed] [Google Scholar]
- 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]
- Spencer S. A., Hammonds R. G., Henzel W. J., Rodriguez H., Waters M. J., Wood W. I. Rabbit liver growth hormone receptor and serum binding protein. Purification, characterization, and sequence. J Biol Chem. 1988 Jun 5;263(16):7862–7867. [PubMed] [Google Scholar]
- 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]
- Yarden Y., Escobedo J. A., Kuang W. J., Yang-Feng T. L., Daniel T. O., Tremble P. M., Chen E. Y., Ando M. E., Harkins R. N., Francke U. Structure of the receptor for platelet-derived growth factor helps define a family of closely related growth factor receptors. Nature. 1986 Sep 18;323(6085):226–232. doi: 10.1038/323226a0. [DOI] [PubMed] [Google Scholar]