Ectopic recombination within homologous immunoglobulin mu gene constant regions in a mouse hybridoma cell line.

Baker, Mark D.; Read, Leah R.

doi:10.1128/mcb.12.10.4422

Cited by 15 publications

(17 citation statements)

References 69 publications

(67 reference statements)

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“…Yet nonhomologous joining of different chromosomes has been documented in translocations, suggesting that DNA sequences at ectopic sites can interact with each other (e.g., Wang et al 1997). In addition, at least in some instances in the mouse, ectopic recombination can occur at a detectable frequency (Baker and Read 1992;Murti et al 1994).…”

mentioning

confidence: 99%

Double-strand break repair by interchromosomal recombination: suppression of chromosomal translocations

Richardson¹,

Moynahan²,

Jasin³

1998

Genes Dev.

399

310

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To directly determine whether recombinational repair of double-strand breaks (DSBs) can occur between heterologous chromosomes and lead to chromosomal rearrangements in mammalian cells, we employed an ES cell system to analyze recombination between repeats on heterologous chromosomes. We found that recombination is induced at least 1000-fold following the introduction of a DSB in one repeat. Most (98%) recombinants repaired the DSB by gene conversion in which a small amount of sequence information was transferred from the unbroken chromosome onto the broken chromosome. The remaining recombinants transferred a larger amount of information, but still no chromosomal aberrations were apparent. Thus, mammalian cells are capable of searching genome-wide for sequences that are suitable for DSB repair. The lack of crossover events that would have led to translocations supports a model in which recombination is coupled to replication.

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mentioning

confidence: 99%

Double-strand break repair by interchromosomal recombination: suppression of chromosomal translocations

Richardson¹,

Moynahan²,

Jasin³

1998

Genes Dev.

399

310

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“…The transformants were screened by Southern blot analysis to identify those bearing the ectopic wild-type donor C region in low copy numbers at (usually) a single independent chromosomal position as described in the Table 1 footnotes. In the transformants, homologous recombination between the ectopic donor wild-type EcoRI C region fragment and the recipient mutant chromosomal gene was detected by the generation of TNP-specific PFC as detailed previously (4,5). The results suggested a noticeable trend toward poorer correction of the larger chromosomal deletions.…”

Section: Resultsmentioning

confidence: 79%

“…We have previously described the system for detecting ectopic homologous recombination in murine hybridomas (5). In brief, it is based upon the ability to detect normal cytolytic, TNP-specific IgM production following homologous recombination between an ectopic wildtype C region donor sequence and the recipient mutant C region of the haploid chromosomal immunoglobulin gene.…”

Section: Resultsmentioning

confidence: 99%

“…Recombinant cells were detected as PFC in a sensitive TNP-specific plaque assay (4,5). Molecular characterization of several recombinants in previous studies suggested that gene conversion was an important mechanism of recombination between dispersed C sequences in the hybridomas (5).…”

Section: Discussionmentioning

confidence: 99%

“…Our laboratory has been investigating homologous recombination in mammalian somatic (murine hybridoma) cells (3)(4)(5)(6)(7). Our assay detects homologous recombination between a donor wild-type immunoglobulin gene constant (C) region and the mutant C region of the haploid, chromosomal immunoglobulin gene in hybridomas which serves as the recipient sequence for recombination.…”

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confidence: 99%

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Requirements for Ectopic Homologous Recombination in Mammalian Somatic Cells

Baker

Read

Beatty

et al. 1996

Molecular and Cellular Biology

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Ectopic recombination occurs between DNA sequences that are not in equivalent positions on homologous chromosomes and has beneficial as well as potentially deleterious consequences for the eukaryotic genome. In the present study, we have examined ectopic recombination in mammalian somatic (murine hybridoma) cells in which a deletion in the gene constant (C) region of the endogenous chromosomal immunoglobulin gene is corrected by using as a donor an ectopic wild-type C region. Ectopic recombination restores normal immunoglobulin M production in hybridomas. We show that (i) chromosomal gene deletions of 600 bp and 4 kb are corrected less efficiently than a deletion of only 2 bp, (ii) the minimum amount of homology required to mediate ectopic recombination is between 1.9 and 4.3 kb, (iii) the frequency of ectopic recombination does not depend on donor copy number, and (iv) the frequency of ectopic recombination in hybridoma lines in which the donor and recipient C regions are physically connected to each other on the same chromosome can be as much as 4 orders of magnitude higher than it is for the same sequences located on homologous or nonhomologous chromosomes. The results are discussed in terms of a model for ectopic recombination in mammalian somatic cells in which the scanning mechanism that is used to locate a homologous partner operates preferentially in cis.Homologous recombination is the process by which genetic information is exchanged between DNA duplexes and can occur by either gene conversion or crossing over. Gene conversion has the property of transferring genetic information in a nonreciprocal manner, while single reciprocal crossover leads to changes in the linkage relationship between genes or groups of genes. Homologous recombination generates new gene combinations that provide the basis for species diversification. It is thought to have played a fundamental role in genome organization and in the maintenance of homogeneity among repeated sequences and members of multigene families (8,15,16,25,56,59). As well, it can correct errors of replication and other forms of DNA damage (24, 27). However, recombination can also lead to altered gene function, altered expression of genes, or the loss of genes. These genetic alterations are implicated in the development of several human cancers (32, 46) and other abnormalities (49, 78).Our laboratory has been investigating homologous recombination in mammalian somatic (murine hybridoma) cells (3-7). Our assay detects homologous recombination between a donor wild-type immunoglobulin gene constant (C) region and the mutant C region of the haploid, chromosomal immunoglobulin gene in hybridomas which serves as the recipient sequence for recombination. Homologous recombination corrects the mutation in the recipient chromosomal gene and restores trinitrophenyl (TNP)-specific immunoglobulin M (IgM) production in mutant cells. Hybridomas making normal TNP-specific IgM are detected as plaque-forming cells (PFC) in a sensitive, TNP-specific plaque assay (4).In a previous...

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Interchromosomal recombination is suppressed in mammalian somatic cells.

et al. 1995

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Homologous recombination occurs intrachromosomally as well as interchromosomally, both in mitotic (somatic) cells as well as meiotically in the germline. These different processes can serve very different purposes in maintaining the integrity of the organism and in enhancing diversity in the species. As shown here, comparison of the frequencies of intra‐ and interchromosomal recombination in meiotic and mitotic cells of both mouse and yeast argues that interchromosomal recombination is particularly low in mitotic cells of metazoan organisms. This result in turn suggests that the recombination machinery of metazoa might be organized to avoid the deleterious effects of homozygotization in somatic cells while still deriving the benefits of species diversification and of DNA repair.

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Ectopic recombination within homologous immunoglobulin mu gene constant regions in a mouse hybridoma cell line.

Cited by 15 publications

References 69 publications

Double-strand break repair by interchromosomal recombination: suppression of chromosomal translocations

Double-strand break repair by interchromosomal recombination: suppression of chromosomal translocations

Requirements for Ectopic Homologous Recombination in Mammalian Somatic Cells

Interchromosomal recombination is suppressed in mammalian somatic cells.

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