Characterization of mRAD18Sc, a Mouse Homolog of the Yeast Postreplication Repair Gene RAD18

Laan, Roald van der; Roest, Henk P.; Hoogerbrugge, Jos W.; Smit, E. M. E.; Slater, Rosalyn; Baarends, Willy M.; Hoeijmakers, Jan H.J.; Grootegoed, J. Anton

doi:10.1006/geno.2000.6220

Cited by 38 publications

(29 citation statements)

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“…Our laboratory has identified two mouse homologs of the ubiquitinconjugating enzyme RAD6, HR6a/Ube2a and HR6b/Ube2b (Roest et al, 1996;Kwon et al, 2001), and recently we also cloned Rad18 Sc , the mouse homolog of the ubiquitin ligase RAD18 (van der Laan et al, 2000). The Rad18 Sc gene is ubiquitously expressed in mouse tissues, but the highest level of mRNA expression is found in testis (van der Laan et al, 2000). To obtain insight into possible spermatogenic aspects of RDB and about Rad18 Sc functions in particular, we have analyzed the expression of Rad18 Sc during spermatogenesis using different mouse models.…”

Section: Introductionmentioning

confidence: 99%

“…High expression of Rad18 Sc mRNA has been detected in primary spermatocytes, in the prophase of the first meiotic division (van der Laan et al, 2000). Following DNA replication in preleptotene spermatocytes, which is the spermatogenic final round of DNA replication, the relatively long meiotic prophase I can be subdivided into four phases: leptotene, zygotene, pachytene and diplotene.…”

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Ubiquitin ligase Rad18Sc localizes to the XY body and to other chromosomal regions that are unpaired and transcriptionally silenced during male meiotic prophase

Laan

Uringa

Wassenaar

et al. 2004

Journal of Cell Science

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Section: Introductionmentioning

confidence: 99%

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Ubiquitin ligase Rad18Sc localizes to the XY body and to other chromosomal regions that are unpaired and transcriptionally silenced during male meiotic prophase

Laan

Uringa

Wassenaar

et al. 2004

Journal of Cell Science

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“…The human homolog of this E3 enzyme, hRAD18Sc, has also been shown to interact with HR6A and HR6B (55). We have recently identified the gene encoding the mouse homolog mRAD18Sc, and the expression of this gene was shown to be highly elevated in the testis, in particular in spermatocytes (49).…”

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Loss of HR6B Ubiquitin-Conjugating Activity Results in Damaged Synaptonemal Complex Structure and Increased Crossing-Over Frequency during the Male Meiotic Prophase

Baarends¹,

Wassenaar²,

Hoogerbrugge³

et al. 2003

Molecular and Cellular Biology

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The ubiquitin-conjugating enzymes HR6A and HR6B are the two mammalian homologs of Saccharomyces cerevisiae RAD6. In yeast, RAD6 plays an important role in postreplication DNA repair and in sporulation. HR6B knockout mice are viable, but spermatogenesis is markedly affected during postmeiotic steps, leading to male infertility. In the present study, increased apoptosis of HR6B knockout primary spermatocytes was detected during the first wave of spermatogenesis, indicating that HR6B performs a primary role during the meiotic prophase. Detailed analysis of HR6B knockout pachytene nuclei showed major changes in the synaptonemal complexes. These complexes were found to be longer. In addition, we often found depletion of synaptonemal complex proteins from near telomeric regions in the HR6B knockout pachytene nuclei. Finally, we detected an increased number of foci containing the mismatch DNA repair protein MLH1 in these nuclei, reflecting a remarkable and consistent increase (20 to 25%) in crossing-over frequency. The present findings reveal a specific requirement for the ubiquitin-conjugating activity of HR6B in relation to dynamic aspects of the synaptonemal complex and meiotic recombination in spermatocytes.Ubiquitin is present in all cells, and the ubiquitin system is involved in different essential cellular processes such as cell division, responses to stress, and apoptosis. Protein ubiquitination occurs through the activities of ubiquitin activatingenzymes (E1), ubiquitin-conjugating enzymes (E2), and ubiquitin-ligating enzymes (E3) (52). Polyubiquitination usually targets a substrate for degradation by a multisubunit structure called the proteasome. Ubiquitination, and in particular monoubiquitination, may also serve other purposes, such as activation or inactivation of transcription factors (12), internalization of transmembrane receptors (22, 52), and alteration of chromatin structure through stable ubiquitination of histones (13, 36). The genome encodes only very few E1 enzymes (1 or 2), and the diverse functions of the ubiquitin system are brought about by 10 to 20 different E2s and an even greater variation of E3 enzymes (52). The ubiquitin-conjugating E2 enzyme HR6B is essential for male fertility in the mouse (41).HR6B is one of the two mammalian homologs of the Saccharomyces cerevisiae E2 enzyme named RAD6/UBC2 (24). The other mammalian RAD6 homolog, HR6A, shows 96% amino acid identity to HR6B. Between mouse and human enzymes, the identity is 100%. The mouse and human HR6B genes are autosomal, whereas HR6A is located on the X chromosome in both species (24,41). RAD6 in yeast is essential for sporulation but is also involved in many other processes, as illustrated by the pleiotropic phenotype of RAD6-null mutants (27). These mutants are defective in a specific DNA damage response pathway named postreplication repair. Furthermore, there are defects in gene silencing and changes in mitotic homologous recombination frequency (10,23,44). The precise mechanism of the role of RAD6 in yeast meiosis and sporulat...

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“…Genetic studies of Saccharomyces cerevisiae suggest that all known components of TLS belong to the epistasis group of the RAD6-RAD18 genes (21,22), which are conserved from yeast to mammalian cells (23)(24)(25)(26)(27). The Rad6 protein is one of the ubiquitin-conjugating enzymes (E2s) and forms a tight complex with the Rad18 protein, which may have E3 ubiquitin ligase activity (28,29 lymphocyte line DT40 (30).…”

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Involvement of Vertebrate Polκ in Rad18-independent Postreplication Repair of UV Damage

Okada¹,

Sonoda²,

Yamashita³

et al. 2002

Journal of Biological Chemistry

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DNA damage, which is left unrepaired by excision repair pathways, often blocks replication, leading to lesions such as breaks and gaps on the sister chromatids. These lesions may be processed by either homologous recombination (HR) repair or translesion DNA synthesis (TLS). Vertebrate Pol belongs to the DNA polymerase Y family, as do most TLS polymerases. However, the role for Pol in vertebrate cells is unclear because of the lack of reverse genetic studies. Here, we generated cells deficient in Pol (pol cells) from the chicken B lymphocyte line DT40. Although purified Pol is unable to bypass ultraviolet (UV) damage, pol cells exhibited increased UV sensitivity, and the phenotype was suppressed by expression of human and chicken Pol, suggesting that Pol is involved in TLS of UV photoproduct. Defects in both Pol and Rad18, which regulates TLS in yeast, in DT40 showed an additive effect on UV sensitivity. Interestingly, the level of sister chromatid exchange, which reflects HR-mediated repair, was elevated in normally cycling pol cells. This implies functional redundancy between HR and Pol in maintaining chromosomal DNA. In conclusion, vertebrate Pol is involved in Rad18-independent TLS of UV damage and plays a role in maintaining genomic stability.A wide range of potential insult to the genomic DNA is contributed not only by the environment, but also by cellular activities. DNA damages, which are left unrepaired by excision repair pathways, may arrest DNA replication, leading to breaks and gaps on the sister chromatids. These lesions are processed by two major postreplication repair pathways: homologous recombination (HR) 1 repair and translesion DNA synthesis (TLS) (1). TLS fills a daughter strand gap that encompasses a DNA damage on the template strand by employing a number of specialized DNA polymerases (Pols) (reviewed in Refs. 2-4). On the other hand, HR functions in processing both gaps and breaks by facilitating recombination between damaged sister chromatids with the other intact ones. Thus, daughter-strand gaps can be processed by both TLS and HR in postreplicational repair.Human cells contain four Y family DNA polymerases, namely Pol (Rad30A), Pol (Rad30B), Pol (DinB1), and Rev1 (5-7). A number of biochemical studies have demonstrated that Y family DNA polymerases can bypass specific lesions on the template strand (8 -12), whereas in vivo function of these polymerases are as yet unclear except for Pol. Pol is mutated in a variant form of xeroderma pigmentosum (XP-V) (13, 14), which is characterized by predisposition to skin cancer and elevated UV sensitivity.

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Characterization of mRAD18Sc, a Mouse Homolog of the Yeast Postreplication Repair Gene RAD18

Cited by 38 publications

References 46 publications

Ubiquitin ligase Rad18Sc localizes to the XY body and to other chromosomal regions that are unpaired and transcriptionally silenced during male meiotic prophase

Ubiquitin ligase Rad18Sc localizes to the XY body and to other chromosomal regions that are unpaired and transcriptionally silenced during male meiotic prophase

Loss of HR6B Ubiquitin-Conjugating Activity Results in Damaged Synaptonemal Complex Structure and Increased Crossing-Over Frequency during the Male Meiotic Prophase

Involvement of Vertebrate Polκ in Rad18-independent Postreplication Repair of UV Damage

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