In Situ Visualization of DNA Double-Strand Break Repair in Human Fibroblasts

Nelms, Benjamin E.; Maser, Richard S.; Mackay, J. F. G.; Lagally, M. G.; Petrini, John H.J.

doi:10.1126/science.280.5363.590

Cited by 457 publications

(297 citation statements)

References 42 publications

(8 reference statements)

Supporting

Mentioning

276

Contrasting

Unclassified

Order By: Relevance

“…In previous studies, it was observed that XP-V cells with impaired p53 employed an MRNdependent recombination pathway in response to UV damage [17,25,27,28], implying DSBs are generated during replication arrest [49,[52][53][54]. We observed that p53 suppression leads to a similar increase in the frequency of normal and XP-V cells with Mre11 foci, even without UV irradiation.…”

Section: Discussionsupporting

confidence: 63%

p53 suppression overwhelms DNA polymerase η deficiency in determining the cellular UV DNA damage response

et al. 2007

View full text Add to dashboard Cite

Xeroderma pigmentosum variant (XP-V) cells lack the damage-specific DNA polymerase η and have normal excision repair but show defective DNA replication after UV irradiation. Previous studies using cells transformed with SV40 or HPV16 (E6/E7) suggested that the S-phase response to UV damage is altered in XP-V cells with non-functional p53. To investigate the role of p53 directly we targeted p53 in normal and XP-V fibroblasts using short hairpin RNA. The shRNA reduced expression of p53, and the downstream cell cycle effector p21, in control and UV irradiated cells. Cells accumulated in late S phase after UV, but after down-regulation of p53 they accumulated earlier in S. Cells in which p53 was inhibited showed ongoing genomic instability at the replication fork. Cells exhibited high levels of UV induced S-phase γH2Ax phosphorylation representative of exposed single strand regions of DNA and foci of Mre11/Rad50/Nbs1 representative of double strand breaks. Cells also showed increased variability of genomic copy numbers after long-term inhibition of p53. Inhibition of p53 expression dominated the DNA damage response. Comparison with earlier results indicates that in virally transformed cells cellular targets other than p53 play important roles in the UV DNA damage response.

show abstract

Section: Discussionsupporting

confidence: 63%

p53 suppression overwhelms DNA polymerase η deficiency in determining the cellular UV DNA damage response

et al. 2007

View full text Add to dashboard Cite

show abstract

“…These processes may be conserved, as broken DSB ends can exhibit limited or pronounced mobility in mammals [63][64][65][66] . Thus, it will be important to determine whether motor complexes contribute to DSB repair in different organisms including human.…”

Section: Discussionmentioning

confidence: 99%

Perinuclear tethers license telomeric DSBs for a broad kinesin- and NPC-dependent DNA repair process

et al. 2015

View full text Add to dashboard Cite

DNA double-strand breaks (DSBs) are often targeted to nuclear pore complexes (NPCs) for repair. How targeting is achieved and the DNA repair pathways involved in this process remain unclear. Here, we show that the kinesin-14 motor protein complex (Cik1-Kar3) cooperates with chromatin remodellers to mediate interactions between subtelomeric DSBs and the Nup84 nuclear pore complex to ensure cell survival via break-induced replication (BIR), an error-prone DNA repair process. Insertion of a DNA zip code near the subtelomeric DSB site artificially targets it to NPCs hyperactivating this repair mechanism. Kinesin-14 and Nup84 mediate BIR-dependent repair at non-telomeric DSBs whereas perinuclear telomere tethers are only required for telomeric BIR. Furthermore, kinesin-14 plays a critical role in telomerase-independent telomere maintenance. Thus, we uncover roles for kinesin and NPCs in DNA repair by BIR and reveal that perinuclear telomere anchors license subtelomeric DSBs for this error-prone DNA repair mechanism.

show abstract

“…The gene product defective in NBS, p95 or nibrin, is a component of the large multiprotein complex containing hRad50 and hMre11, nuclear proteins implicated in the NHEJ recombinational DNA repair pathway (Varon et al, 1998;Carney et al, 1998;Dolganov et al, 1996;Featherstone and Jackson, 1998). In response to IR, hMre11 and hRad50 form nuclear foci in close proximity to DSBs and the formation of these foci is markedly reduced in both NBS and AT cells (Maser et al, 1997;Dolganov et al, 1996;Carney et al, 1998;Nelms et al, 1998). Interestingly, AT cells form unusually high numbers of foci containing the homologous recombinational repair protein, hRad51 (Maser et al, 1997).…”

Section: At Phenotypementioning

confidence: 99%