Mutation of cysteine-88 in the Saccharomyces cerevisiae RAD6 protein abolishes its ubiquitin-conjugating activity and its various biological functions - PubMed (original) (raw)
Mutation of cysteine-88 in the Saccharomyces cerevisiae RAD6 protein abolishes its ubiquitin-conjugating activity and its various biological functions
P Sung et al. Proc Natl Acad Sci U S A. 1990 Apr.
Abstract
The RAD6 gene of Saccharomyces cerevisiae is required for DNA repair, DNA damage-induced mutagenesis, and sporulation. RAD6 protein is a ubiquitin-conjugating enzyme (E2) that has been shown to attach multiple molecules of ubiquitin to histones H2A and H2B. We have now examined whether the E2 activity of RAD6 is involved in its various biological functions. Since the formation of a thioester adduct between E2 and ubiquitin is necessary for E2 activity, the single cysteine residue (Cys-88) present in RAD6 was changed to alanine or valine. The mutant proteins were overproduced in yeast cells and purified to near homogeneity. We show that the rad6 Ala-88 and rad6 Val-88 mutant proteins lack the capacity for thioester formation with ubiquitin and, as a consequence, are totally devoid of any E2 activity. The rad6 Ala-88 and rad6 Val-88 mutations confer a defect in DNA repair, mutagenesis, and sporulation equivalent to that in the rad6 null allele. We suggest that the biological functions of RAD6 require its E2 activity.
Similar articles
- Stable ester conjugate between the Saccharomyces cerevisiae RAD6 protein and ubiquitin has no biological activity.
Sung P, Prakash S, Prakash L. Sung P, et al. J Mol Biol. 1991 Oct 5;221(3):745-9. doi: 10.1016/0022-2836(91)80169-u. J Mol Biol. 1991. PMID: 1658333 - A ubiquitin carrier protein from wheat germ is structurally and functionally similar to the yeast DNA repair enzyme encoded by RAD6.
Sullivan ML, Vierstra RD. Sullivan ML, et al. Proc Natl Acad Sci U S A. 1989 Dec;86(24):9861-5. doi: 10.1073/pnas.86.24.9861. Proc Natl Acad Sci U S A. 1989. PMID: 2557633 Free PMC article. - The RAD6 protein of Saccharomyces cerevisiae polyubiquitinates histones, and its acidic domain mediates this activity.
Sung P, Prakash S, Prakash L. Sung P, et al. Genes Dev. 1988 Nov;2(11):1476-85. doi: 10.1101/gad.2.11.1476. Genes Dev. 1988. PMID: 2850263 - The structure and function of RAD6 and RAD18 DNA repair genes of Saccharomyces cerevisiae.
Prakash L. Prakash L. Genome. 1989;31(2):597-600. doi: 10.1139/g89-111. Genome. 1989. PMID: 2698834 Review. - The RAD6 gene of yeast: a link between DNA repair, chromosome structure and protein degradation?
Siede W. Siede W. Radiat Environ Biophys. 1988;27(4):277-86. doi: 10.1007/BF01209756. Radiat Environ Biophys. 1988. PMID: 2852379 Review.
Cited by
- Ubiquitination Occurs in the Mitochondrial Matrix by Eclipsed Targeted Components of the Ubiquitination Machinery.
Zhang Y, Karmon O, Das K, Wiener R, Lehming N, Pines O. Zhang Y, et al. Cells. 2022 Dec 17;11(24):4109. doi: 10.3390/cells11244109. Cells. 2022. PMID: 36552873 Free PMC article. - RAD6 inhibition enhances paclitaxel sensitivity of triple negative breast cancer cells by aggravating mitotic spindle damage.
Haynes BM, Cunningham K, Shekhar MPV. Haynes BM, et al. BMC Cancer. 2022 Oct 18;22(1):1073. doi: 10.1186/s12885-022-10119-z. BMC Cancer. 2022. PMID: 36258187 Free PMC article. - Itch regulation of innate and adaptive immune responses in mice and humans.
Field NS, Moser EK, Oliver PM. Field NS, et al. J Leukoc Biol. 2020 Jul;108(1):353-362. doi: 10.1002/JLB.3MIR0320-272R. Epub 2020 Apr 30. J Leukoc Biol. 2020. PMID: 32356405 Free PMC article. Review. - RAD6B is a major mediator of triple negative breast cancer cisplatin resistance: Regulation of translesion synthesis/Fanconi anemia crosstalk and BRCA1 independence.
Haynes B, Gajan A, Nangia-Makker P, Shekhar MP. Haynes B, et al. Biochim Biophys Acta Mol Basis Dis. 2020 Jan 1;1866(1):165561. doi: 10.1016/j.bbadis.2019.165561. Epub 2019 Oct 19. Biochim Biophys Acta Mol Basis Dis. 2020. PMID: 31639439 Free PMC article. - Endoplasmic reticulum-localized UBC34 interaction with lignin repressors MYB221 and MYB156 regulates the transactivity of the transcription factors in Populus tomentosa.
Zheng L, Chen Y, Ding D, Zhou Y, Ding L, Wei J, Wang H. Zheng L, et al. BMC Plant Biol. 2019 Mar 12;19(1):97. doi: 10.1186/s12870-019-1697-y. BMC Plant Biol. 2019. PMID: 30866808 Free PMC article.
References
- Nature. 1970 Aug 15;227(5259):680-5 - PubMed
- Mutat Res. 1974 Sep;24(3):281-92 - PubMed
- Cell. 1982 May;29(1):11-22 - PubMed
- Genetics. 1974 Dec;78(4):1101-18 - PubMed
- Proc Natl Acad Sci U S A. 1987 Dec;84(24):8951-5 - PubMed
Publication types
MeSH terms
Substances
LinkOut - more resources
Full Text Sources
Molecular Biology Databases