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DNA RepairCornerstone Guide

Homologous Recombination Repair

Homologous recombination (HR) repairs DNA double-strand breaks primarily in S/G2, when a sister chromatid is available as a template. It is generally higher fidelity than end-joining pathways, although no repair process is literally error-free. BRCA1, PALB2, BRCA2 and RAD51 act at different steps. Loss of HR function can leave genomic scars and may create PARP-inhibitor vulnerability, but 'BRCAness', HRD assay results and treatment benefit are related rather than interchangeable concepts.

Quick Answer

HR begins with DSB detection by MRN, which recruits ATM and promotes 5′→3′ DNA end resection to generate 3′ single-stranded DNA (ssDNA) overhangs. RPA covers ssDNA to prevent secondary structure formation. BRCA1 (recruited via ubiquitin/RAP80 cascade) bridges to PALB2, which recruits BRCA2. BRCA2 displaces RPA and loads RAD51 monomers onto ssDNA, forming a nucleoprotein filament. The RAD51-ssDNA filament performs strand invasion of the homologous duplex sister chromatid, using it as a template for accurate DNA synthesis before resolution and ligation.

Mechanism Overview

HR begins with DSB detection by MRN, which recruits ATM and promotes 5′→3′ DNA end resection to generate 3′ single-stranded DNA (ssDNA) overhangs. RPA covers ssDNA to prevent secondary structure formation. BRCA1 (recruited via ubiquitin/RAP80 cascade) bridges to PALB2, which recruits BRCA2. BRCA2 displaces RPA and loads RAD51 monomers onto ssDNA, forming a nucleoprotein filament. The RAD51-ssDNA filament performs strand invasion of the homologous duplex sister chromatid, using it as a template for accurate DNA synthesis before resolution and ligation.

Homologous Recombination Repair

DNA Double-Strand BreakMRN · CtIPend resectionRPA · BRCA1ssDNA coatingPALB2 · BRCA2RAD51 loaderRAD51strand invasionTemplate-directed repairPARP inhibitorsolaparib · niraparib
kinase / signallingoncogenetumour suppressorcellular outputinhibits

Step-by-Step Pathway

1
DSB Detection and End Resection

MRN complex detects DSB ends and recruits ATM. ATM phosphorylates CtIP, which together with MRE11 nuclease and BLM/Exo1 helicase/nucleases catalyses 5′→3′ DNA end resection, generating 3′ ssDNA overhangs of hundreds to thousands of nucleotides.

2
RPA Coating and Cell Cycle Restriction

Replication protein A (RPA) coats 3′ ssDNA overhangs, preventing secondary structure and activating ATR kinase. HR is restricted to S/G2 phase by CDK1/2-mediated phosphorylation of CtIP, ensuring the sister chromatid template is present before HR is initiated.

3
BRCA1 and PALB2 Bridge Assembly

BRCA1 (recruited via RNF168 ubiquitin cascade and RAP80) directly binds PALB2 via its coiled-coil domain. PALB2 acts as a molecular bridge to BRCA2, co-localising BRCA2 with the ssDNA substrate. This three-component assembly (BRCA1–PALB2–BRCA2) is the central scaffold of the HR repair complex.

4
RAD51 Filament Loading by BRCA2

BRCA2 (with DSS1) displaces RPA from ssDNA and loads RAD51 monomers in an ATP-dependent reaction, forming a right-handed helical RAD51-ssDNA nucleoprotein filament. This filament has the capacity to search homologous duplex DNA sequences over megabase distances.

5
Strand Invasion and D-Loop Formation

The RAD51-ssDNA filament performs strand invasion of the intact sister chromatid at the homologous sequence, forming a displacement loop (D-loop). The 3′ ssDNA end within the D-loop serves as primer for DNA synthesis using the sister chromatid as error-free template.

6
DNA Synthesis, Resolution, and Ligation

DNA Pol δ/ε extends the invaded strand using the sister chromatid template. After second end capture (double-strand break repair pathway) or dissolution (synthesis-dependent strand annealing, SDSA), Holiday junctions are resolved by GEN1/MUS81-EME1 or dissolved by BLM-TOP3α-RMI1/2, restoring the original sequence with high fidelity.

Disease Relevance

Pathogenic germline variants in BRCA1, BRCA2 and several other repair genes can create inherited cancer predisposition, but risk estimates differ by gene, variant, age, sex and family history. Tumours can also acquire somatic changes or epigenetic silencing that impair HR. A genomic-scar result describes patterns accumulated over the tumour's history; it is not identical to a BRCA variant, does not establish inheritance and may not prove that HR is currently inactive.

Therapeutic Implications

PARP inhibition can exploit vulnerabilities associated with BRCA1/2 loss and other homologous-recombination defects through catalytic inhibition, PARP trapping and replication-associated damage. Regulatory indications differ by tumour, biomarker, germline or somatic status, prior therapy, maintenance setting and combination. An HRD score or ATM alteration alone should not be interpreted as proof of equivalent benefit across these settings.

Common Questions

What is the difference between HR and NHEJ for DSB repair?

Homologous recombination (HR) usually uses a sister chromatid as a template and is most available in S/G2. Non-homologous end joining (NHEJ) ligates DNA ends without a homologous template and operates more broadly across the cell cycle. HR is generally higher fidelity, while end joining can alter sequence at the junction; neither pathway is accurately described as infallible in every context.

What is PARP trapping and why does it differ from catalytic PARP inhibition?

Catalytic PARP inhibition simply reduces PARP enzymatic activity (NAD+ consumption). PARP trapping — the more cytotoxic mechanism of current PARP inhibitors — involves the inhibitor stabilising the PARP1-DNA complex after nick detection, preventing PARP release and creating a physical 'roadblock' on DNA that is lethal when encountered by a replication fork. Talazoparib is the most potent PARP trapper; niraparib and olaparib have intermediate trapping; veliparib has poor trapping despite catalytic inhibition.

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References

  1. 1Moynahan ME, Jasin M (2010). Mitotic homologous recombination maintains genomic stability and suppresses tumorigenesis. Nat Rev Mol Cell Biol. PubMed 20571552
  2. 2Lord CJ, Ashworth A (2017). PARP inhibitors: Synthetic lethality in the clinic. Science. PubMed 28302823
  3. 3Roy R, Chun J, Powell SN (2012). BRCA1 and BRCA2: different roles in a common pathway of genome protection. Nat Rev Cancer. PubMed 22193176
  4. 4Kuchenbaecker KB, et al. (2017). Risks of Breast, Ovarian, and Contralateral Breast Cancer for BRCA1 and BRCA2 Mutation Carriers. JAMA. PubMed 28291904