Cloning and Expression of cDNA Encoding a Human Ubiquitin-Conjugating Enzyme Similar to the Drosophila bendless Gene Product

Yamaguchi, Tomoko; Kim, Nam-Soon; Sekine, Shingo; Seino, Hiroaki; Osaka, Fumio; Yamao, Fumiaki; Kato, Souichiro

doi:10.1093/oxfordjournals.jbchem.a021440

Cited by 38 publications

(18 citation statements)

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“…Over the last few years, a number of mammalian homologs of yeast RDB genes that exhibit a high degree of sequence and functional conservation have been identified, suggesting that RDB mechanisms might be conserved throughout eukaryotic evolution (21,30,33,42,44,47,68,72,(78)(79)(80). However, some remarkable functional differences between yeast and mammalian proteins (8,17,73,76) and the identification of additional Y family DNA polymerases in mammalian species (for a review, see reference 19) forbid simple extrapolation from yeast to mammals.…”

Section: Discussionmentioning

confidence: 99%

The Ubiquitin-Conjugating DNA Repair Enzyme HR6A Is a Maternal Factor Essential for Early Embryonic Development in Mice

Roest

Baarends

Wit³

et al. 2004

Molecular and Cellular Biology

108

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The Saccharomyces cerevisiae RAD6 protein is required for a surprising diversity of cellular processes, including sporulation and replicational damage bypass of DNA lesions. In mammals, two RAD6-related genes, HR6A and HR6B, encode highly homologous proteins. Here, we describe the phenotype of cells and mice deficient for the mHR6A gene. Just like mHR6B knockout mouse embryonic fibroblasts, mHR6A-deficient cells appear to have normal DNA damage resistance properties, but mHR6A knockout male and female mice display a small decrease in body weight. The necessity for at least one functional mHR6A (X-chromosomal) or mHR6B (autosomal) allele in all somatic cell types is supported by the fact that neither animals lacking both proteins nor females with only one intact mHR6A allele are viable. In striking contrast to mHR6B knockout males, which show a severe spermatogenic defect, mHR6A knockout males are normally fertile. However, mHR6A knockout females fail to produce offspring despite a normal ovarian histology and ovulation. The absence of mHR6A in oocytes prevents development beyond the embryonic two-cell stage but does not result in an aberrant methylation pattern of histone H3 at this early stage of mouse embryonic development. These observations support redundant but dose-dependent roles for HR6A and HR6B in somatic cell types and germ line cells in mammals.Repair of damaged DNA requires a number of complementary, partially overlapping mechanisms, including mismatch repair, nucleotide and base excision repair, and repair pathways that deal with double-strand breaks. S-phase arrest caused by unrepaired DNA damage is prevented by a mechanism known as postreplication repair, which we prefer to rename replicational damage bypass (RDB). This bypass mechanism allows replication across lesions instead of repairing lesions (20,26).Based on the mutant phenotype, RAD6 is one of the key players in RDB in yeast. rad6 mutants are extremely sensitive to a wide range of DNA-damaging agents and show an increased spontaneous mutation frequency and a loss of DNA damage-induced mutagenesis (41,46). In addition, RAD6 is also involved in a number of other cellular processes in yeast, including the degradation of proteins via the N-end rule, retrotransposition, gene silencing, meiosis, and sporulation (14, 15, 28, 52). RAD6 was identified as a ubiquitin-conjugating enzyme (29), and all of the functions of RAD6 in yeast depend on its ability to transfer ubiquitin molecules to a protein substrate (66). The ubiquitin-conjugating pathway requires the consecutive activity of ubiquitin-activating (E1), -conjugating (E2), and -ligating (E3) enzymes and results in mono-or polyubiquitination of substrate proteins. The ubiquitination serves as a signal for modification, activation, or proteolytic degradation via the 26S proteasome. The identification of 11 ubiquitinconjugating enzymes in yeast and at least 20 genes encoding such proteins in the human genome, as well as the existence of a vast number of E3s, suggests that high substrate specific...

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Section: Discussionmentioning

confidence: 99%

The Ubiquitin-Conjugating DNA Repair Enzyme HR6A Is a Maternal Factor Essential for Early Embryonic Development in Mice

Roest

Baarends

Wit³

et al. 2004

Molecular and Cellular Biology

108

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“…Cleavage results in the separation of GST from Mms2 or Ubc13, both of which contain Gly-Pro-Leu-Gly-Ser appended to the N terminus of the first codon. The DNA sequences were derived from plasmid-borne cloned versions for Mms2 (33) and Ubc13 (34), respectively. The DNA sequences of the human UBC13 and MMS2 coding regions were verified by sequencing each recombinant plasmid.…”

Section: Methodsmentioning

confidence: 99%

“…The full-length human MMS2 cDNA is able to complement the yeast mms2 mutant, suggesting that human Mms2 is indeed a functional homolog of yeast Mms2. In addition, a human UBC13 cDNA was isolated as a homolog of the Drosophila melanogaster bendless gene (33). Human UBC13 cDNA is also able to complement the yeast Ubc13 mutant.…”

mentioning

confidence: 99%

Noncovalent Interaction between Ubiquitin and the Human DNA Repair Protein Mms2 Is Required for Ubc13-mediated Polyubiquitination

McKenna¹,

Spyracopoulos²,

Moraes³

et al. 2001

Journal of Biological Chemistry

125

160

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Ubiquitin-conjugating enzyme variants share significant sequence similarity with typical E2 (ubiquitin-conjugating) enzymes of the protein ubiquitination pathway but lack their characteristic active site cysteine residue. The MMS2 gene of Saccharomyces cerevisiae encodes one such ubiquitin-conjugating enzyme variant that is involved in the error-free DNA postreplicative repair pathway through its association with Ubc13, an E2. The Mms2-Ubc13 heterodimer is capable of linking ubiquitin molecules to one another through an isopeptide bond between the C terminus and Lys-63. Using highly purified components, we show here that the human forms of Mms2 and Ubc13 associate into a heterodimer that is stable over a range of conditions. The ubiquitin-thiol ester form of the heterodimer can be produced by the direct activation of its Ubc13 subunit with E1 (ubiquitin-activating enzyme) or by the association of Mms2 with the Ubc13-ubiquitin thiol ester. The activated heterodimer is capable of transferring its covalently bound ubiquitin to Lys-63 of an untethered ubiquitin molecule, resulting in diubiquitin as the predominant species. In 1 H 15 N HSQC ( 1 H 15 N heteronuclear single quantum coherence) NMR experiments, we have mapped the surface determinants of tethered and untethered ubiquitin that interact with Mms2 and Ubc13 in both their monomeric and dimeric forms. These results have identified a surface of untethered ubiquitin that interacts with Mms2 in the monomeric and heterodimeric form. Furthermore, the C-terminal tail of ubiquitin does not participate in this interaction. These results suggest that the role of Mms2 is to correctly orient either a target-bound or untethered ubiquitin molecule such that its Lys-63 is placed proximally to the C terminus of the ubiquitin molecule that is linked to the active site of Ubc13.

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“…Two Unigene clusters lie within this interval, one at the centromeric end of BAC 52 and the other between Nmu and Clock. Finally, pseudogenes for platelet-activating factor acetylhydrolase Ib alpha subunit (Pafah1b1-ps2), adenopolyposis coli (APC), and ubiquitin-conjugating enzyme homolog (bendless) exist in the 204-kb region (Joslyn et al 1991;Su et al 1992;Yamaguchi et al 1996;Peterfy et al 1998). The Pafah1b1-ps2 and bendless pseudogenes are full-length, while the APC pseudogene represents 1240 bases near the 3Ј end of the functional APC gene.…”

Section: Shotgun Sequencing and Finishing Of Two Bacterial Artificialmentioning

confidence: 99%

The Mouse Clock Locus: Sequence and Comparative Analysis of 204 Kb from Mouse Chromosome 5

Wilsbacher

Sangoram²,

Antoch³

et al. 2000

Genome Research

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Cloning and Expression of cDNA Encoding a Human Ubiquitin-Conjugating Enzyme Similar to the Drosophila bendless Gene Product

Cited by 38 publications

References 0 publications

The Ubiquitin-Conjugating DNA Repair Enzyme HR6A Is a Maternal Factor Essential for Early Embryonic Development in Mice

The Ubiquitin-Conjugating DNA Repair Enzyme HR6A Is a Maternal Factor Essential for Early Embryonic Development in Mice

Noncovalent Interaction between Ubiquitin and the Human DNA Repair Protein Mms2 Is Required for Ubc13-mediated Polyubiquitination

The Mouse Clock Locus: Sequence and Comparative Analysis of 204 Kb from Mouse Chromosome 5

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