Human and mouse homologs of the Saccharomyces cerevisiaeRAD54 DNA repair gene: evidence for functional conservation

Kanaar, Roland; Troelstra, Christine; Swagemakers, Sigrid; Essers, Jeroen; Smit, Bep; Franssen, Jan Huib; Pastink, Albert; Bezzubova, Olga; Buerstedde, Jean Marie; Clever, Beate; Heyer, Wolf Dietrich; Hoeijmakers, Jan

doi:10.1016/s0960-9822(02)00606-1

Cited by 144 publications

(107 citation statements)

References 49 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…19 Vertebrate homologues of these genes have been recently identified. [20][21][22] The high degree of sequence conservation from yeast to man and the similarity of in vitro biochemical function suggested that the vertebrate homologues were also involved in homologous recombination. RAD51, a eukaryotic homologue of E. coli recA, seems to play a central role in the process with its DNA pairing and strand exchange activities.…”

Section: Correspondence: Acg Porter This Paper Is Dedicated To the Mementioning

confidence: 99%

Gene targeting is enhanced in human cells overexpressing hRAD51

Yáñez

Porter

1999

Gene Ther

View full text Add to dashboard Cite

The ideal therapy for single gene disorders would be repair approach, we have overexpressed the gene encoding of the mutated disease genes. Homologous recombination hRAD51, a protein with homologous DNA pairing and is one of several cellular mechanisms for the repair of DNA strand exchange activities, in human cells and measured damage. Recombination between exogenous DNA and its effect on gene targeting. We report a two-to three-fold homologous chromosomal loci (gene targeting) can be increase in gene targeting, and enhanced resistance to used to repair an endogenous gene, but the low efficiency ionising radiation in hRAD51-overexpressing cells with no of this process is a serious barrier to its therapeutic potenobvious detrimental effects. These observations provide tial. Recent progress in the isolation and characterisation valuable genetic evidence for the involvement of hRAD51 of mammalian genes and proteins involved in DNA recomin both gene targeting and DNA repair in human cells. Our bination has raised the possibility that the cellular biochemdata also establish overexpression of recombination genes istry of recombination can be manipulated to improve the as a viable approach to improving gene targeting efficiency of gene targeting. As an initial test of this efficiencies.

show abstract

Section: Correspondence: Acg Porter This Paper Is Dedicated To the Mementioning

confidence: 99%

Gene targeting is enhanced in human cells overexpressing hRAD51

Yáñez

Porter

1999

Gene Ther

View full text Add to dashboard Cite

show abstract

“…Hybridization was performed overnight at 658C in 0.5 M NaPO 4 (pH7.2), 1 mM EDTA, 7% SDS and 1% bovine serum albumin, followed by washing three times for 30 min at 658C in 40 mM NaPO 4 (pH 7.2), 1 mM EDTA and 1% SDS. Nucleotide sequence was determined using Sequenase sequencing kit (Amersham) Figure 2 Multiple sequence alignment comparing the predicted helicase motifs of RAD54B to those of hRAD54, S. pombe RAD54 (SpRAD54) and ScRAD54 (Kanaar et al, 1996). Amino acid positions are shown on the left.…”

mentioning

confidence: 99%

“…Furthermore, this sequence is well conserved in the SNF2 superfamily regardless of species and subfamily. Asn at codon 593 is conserved in SNF2 (Laurent et al, 1991), MOT1 (Davis et al, 1992), BRM (Tamkun et al, 1992), ERCC6 (Troelstra et al, 1992), STH1 (Clark et al, 1992), FUN30 (Laurent et al, 1992) and RAD54 (Kanaar et al, 1996) throughout evolution from yeast to human. Asp at codon 418 is also at a conserved position ( Figure 6b).…”

mentioning

confidence: 99%

Mutations of a novel human RAD54 homologue, RAD54B, in primary cancer

et al. 1999

View full text Add to dashboard Cite

Association of breast tumor susceptibility gene products BRCA1 and BRCA2 with the RAD51 recombination protein suggested that cancer could arise through defects in recombination. The identi®cation of NBS1, responsible for Nijmegen breakage syndrome, from the MRE11/RAD50 recombination protein complex also supports this hypothesis. However, our mutation analysis revealed that known members of the RAD52 epistasis group are rarely mutated in human primary cancer. Here we describe the isolation of a novel member of the SNF2 superfamily, characterized with sequence motifs similar to those in DNA and RNA helicases. The gene, designated RAD54B, is signi®cantly homologous to the RAD54 recombination gene. The expression of RAD54B was high in testis and spleen, which are active in meiotic and mitotic recombination. These ®ndings suggest that RAD54B may play an active role in recombination processes in concert with other members of the RAD52 epistasis group. RAD54B maps to human chromosome 8q21.3-q22 in a region associated with cancer-related chromosomal abnormalities. Homozygous mutations at highly conserved positions of RAD54B were observed in human primary lymphoma and colon cancer. These ®ndings suggest that some cancers arise through alterations of the RAD54B function.

show abstract

“…Two Rad54 homologous proteins have been found in different eukaryotic species (29). The human hRad54 protein has 68% similarity with ScRad54 protein.…”

mentioning

confidence: 99%

Effects of Tumor-associated Mutations on Rad54 Functions

Smirnova

Komen

Sung

et al. 2004

Journal of Biological Chemistry

View full text Add to dashboard Cite

Yeast RAD54 gene, a member of the RAD52 epistasis group, plays an important role in homologous recombination and DNA double strand break repair. Rad54 belongs to the Snf2/Swi2 protein family, and it possesses a robust DNA-dependent ATPase activity, uses free energy from ATP hydrolysis to supercoil DNA, and cooperates with the Rad51 recombinase in DNA joint formation. There are two RAD54-homologous genes in human cells, hRAD54 and RAD54B. Mutations in these human genes have been found in tumors. These tumor-associated mutations map to conserved regions of the hRad54 and hRad54B proteins. Here we introduced the equivalent mutations into the Saccharomyces cerevisiae RAD54 gene in an effort to examine the functional consequences of these gene changes. One mutant, rad54 G484R, showed sensitivity to DNA-damaging agents and reduced homologous recombination rates, indicating a loss of function. Even though the purified rad54 G484R mutant protein retained the ability to bind DNA and interact with Rad51, it was nearly devoid of ATPase activity and was similarly defective in DNA supercoiling and D-loop formation. Two other mutants, rad54 N616S and rad54 D442Y, were not sensitive to genotoxic agents and behaved like the wild type allele in homologous recombination assays. Consistent with the mild phenotype associated with the rad54 N616S allele, its encoded protein was similar to wild type Rad54 protein in biochemical attributes. Because dysfunctional homologous recombination gives rise to genome instability, our results are consistent with the premise that tumor-associated mutations in hRad54 and Rad54B could contribute to the tumor phenotype or enhance the genome instability seen in tumor cells.In Saccharomyces cerevisiae, homologous recombination (HR) 1 represents an important means for the repair of DNA double strand breaks and other types of DNA damage and also for restarting stalled replication forks (1-5). HR depends on the use of a homologous DNA molecule as a template to eliminate double strand breaks and other lesions mostly with high fidelity. Impaired HR results in genetic instability and sensitivity to genotoxic agents (6) and can lead to cancer in humans (7). Double strand break repair by HR is dependent on genes of the RAD52 epistasis group, RAD50, RAD51, RAD54, RAD55, RAD57, RAD59, RDH54/TID1, MRE11, and XRS2, in which structure and function have been highly conserved (6). Before repair can occur, the ends of double strand breaks are processed by nucleolytic resection of the 5Ј ends of the breaks to yield ssDNA, which is then bound by Rad51 to form a helical nucleoprotein filament often referred to as the presynaptic filament. Nucleation of Rad51 onto the ssDNA is facilitated by recombination mediators, including Rad52, the Rad55-Rad57 complex, and Rad54 (6,8). Once assembled, the presynaptic filament mediates a search for homology in the homologous dsDNA partner and forms a DNA joint molecule (called D-loop) with the latter. The length of the nascent DNA joint molecule is extended by DNA branch migration (9...

show abstract

Human and mouse homologs of the Saccharomyces cerevisiaeRAD54 DNA repair gene: evidence for functional conservation

Cited by 144 publications

References 49 publications

Gene targeting is enhanced in human cells overexpressing hRAD51

Gene targeting is enhanced in human cells overexpressing hRAD51

Mutations of a novel human RAD54 homologue, RAD54B, in primary cancer

Effects of Tumor-associated Mutations on Rad54 Functions

Contact Info

Product

Resources

About