Abstract

Class switch recombination (CSR) is a region-specific, transcriptionally regulated, nonhomologous recombinational process that is initiated by activation-induced cytidine deaminase (AID). The initial lesions in the switch (S) regions are processed and resolved, leading to a recombination of the two S regions involved. The mechanism involved in the repair and ligation of the broken DNA ends is however still unclear. Here, we describe that switching is less efficient in cells from patients with Mre11 deficiency (Ataxia-Telangiectasia-like disorder, ATLD) and, more importantly, that the switch recombination junctions resulting from the in vivo switching events are aberrant. There was a trend toward an increased usage of microhomology (> or =4 bp) at the switch junctions in both ATLD and Nijmegen breakage syndrome (NBS) patients. However, the DNA ends were not joined as "perfectly" as those from Ataxia-Telangiectasia (A-T) patients and 1-2 bp mutations or insertions were often observed. In switch junctions from ATLD patients, there were fewer base substitutions due to transitions and, most strikingly, the substitutions that occurred most often in controls, C --> T transitions, never occurred at, or close to, the junctions derived from the ATLD patients. In switch junctions from NBS patients, all base substitutions were observed at the G/C nucleotides, and transitions were preferred. These data suggest that the Mre11-Rad50-Nbs1 complex (Mre11 complex) is involved in the nonhomologous end joining pathway in CSR and that Mre11, Nbs1, and protein mutated in ataxia-telangiectasia (ATM) might have both common and independent roles in this process.

Highlights

  • The repair of DNA double-strand breaks (DSB)1 is crucial for the maintenance of genome stability and defects in the cellular response to DSBs have been linked to a number of inherited human cancer-prone syndromes [1, 2]

  • We studied somatic hypermutation (SHM)-like mutations in the S␮ regions in both Ataxia-Telangiectasialike disorder (ATLD) and Nijmegen breakage syndrome (NBS) patients

  • The Mre11 complex is one of the most critical biochemical components involved in DNA DSBs and repair [26, 27, 48]

Read more

Summary

No of S fragments

11.9 Ϯ 2.3 a Mutations in the Mre gene in these patients have been described previously [30, 38]. b Ho, homozygous. c He, heterozygous. Biochemical studies have provided a functional link between the Mre complex and ATM [31,32,33,34,35], suggesting that these factors may be required in a common pathway(s) in DNA repair. The frequency and pattern of mutations in the V region is largely normal, suggesting a role for ATM in CSR, but not in SHM [19, 20]. We studied SHM-like mutations in the S␮ regions in both ATLD and NBS patients. These data may help delineate the DSB repair pathways in CSR and SHM

EXPERIMENTAL PROCEDURES
RESULTS
Perfectly matched short homology
DISCUSSION
TCA G

Talk to us

Join us for a 30 min session where you can share your feedback and ask us any queries you have

Schedule a call

Disclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.