DNA Replication and Repair Mechanisms

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DNA Replication Essentials - The Blueprint Copiers

  • Semiconservative: Each new DNA: one parental, one new strand. Ensures faithful copying.
  • Bidirectional: From origin (Ori), two replication forks proceed in opposite directions.
  • Direction: Synthesis strictly $5' \to 3'$. Leading strand continuous; lagging discontinuous (Okazaki fragments).
  • Core Sequence:
  • Key Enzymes:
    • Helicase: Unwinds DNA (ATP-dependent). 📌 "Heli-case cuts helix."
    • Topoisomerases: Relieve supercoils (e.g., DNA gyrase).
    • Primase: Synthesizes RNA primers for DNA Pol.
    • DNA Polymerase: Synthesizes DNA; proofreads ($3' \to 5'$ exonuclease). (Prok: Pol III; Euk: Pol $\delta, \epsilon$).
    • Ligase: Joins DNA fragments. DNA Replication Fork with Enzymes and Okazaki Fragments

⭐ DNA replication in eukaryotes initiates at multiple origins of replication (ARS - autonomously replicating sequences) per chromosome, unlike prokaryotes with a single origin (OriC).

Prokaryotic vs. Eukaryotic Replication - Cellular Copying Quirks

  • Core Contrast: Prokaryotic replication is simpler, faster, with a single origin; Eukaryotic is complex, slower, featuring multiple origins and telomere maintenance.
FeatureProkaryotesEukaryotes
OriginSingle (oriC)Multiple (ARS in yeast)
DNA PolPol I, II, III (main: Pol III)Pol $\alpha, \delta, \epsilon$ (main: Pol $\delta, \epsilon$)
RateFaster: ~1000 nt/secSlower: ~50-100 nt/sec
Okazaki FragsLonger: ~1000-2000 ntShorter: ~100-200 nt
GenomeCircular, no histonesLinear, histones (chromatin)
TelomeresNoYes (Telomerase)
InitiationDnaA @ oriCOrigin Recognition Complex (ORC) @ origins

⭐ Eukaryotic replication is tightly coupled to the cell cycle, primarily occurring during the S phase, unlike prokaryotes where it can occur continuously.

DNA Damage & Repair Pathways - Code Correction Crew

  • Damage Sources:
    • Endogenous: Depurination, Deamination (C→U), Oxidation (8-oxoG).
    • Exogenous: UV (pyrimidine dimers), Alkylating agents, X-rays (DSBs).
  • Key Repair Systems:
    • Direct Reversal: MGMT (O6-methylguanine).
    • Base Excision Repair (BER): For single base damage (uracil, 8-oxoG).
      • Key Enzymes: Glycosylase, AP Endo, Pol, Ligase. 📌 Go Eat Pizza Later.
    • Nucleotide Excision Repair (NER): For bulky lesions (pyrimidine dimers).
      • Defect → Xeroderma Pigmentosum (XP).
    • Mismatch Repair (MMR): For replication errors. MSH, MLH proteins.
      • Defect → Lynch Syndrome (HNPCC).
    • Double-Strand Break (DSB) Repair:
      • Homologous Recombination (HR): Error-free. S/G2 phase. Proteins: BRCA1/2, RAD51.
      • Non-Homologous End Joining (NHEJ): Error-prone. G1 phase. Proteins: Ku70/80.
    • Translesion Synthesis (TLS): Error-prone bypass polymerases.

⭐ Xeroderma Pigmentosum (XP) results from NER defects, causing extreme UV sensitivity and high skin cancer risk.

DNA damage types and repair pathways

Clinical Correlates & Inhibitors - Healing Code Glitches

  • Repair Defect Syndromes:
    • Xeroderma Pigmentosum (XP): NER defect; UV hypersensitivity, ↑ skin Ca.
    • Lynch Syndrome (HNPCC): MMR (MSH2, MLH1) defect; ↑ colorectal, endometrial Ca.
    • Ataxia-Telangiectasia (AT): ATM gene (NHEJ) defect; ataxia, telangiectasias, immunodeficiency.
    • Fanconi Anemia: DNA crosslink repair defect; pancytopenia, congenital anomalies.
  • Key Replication Inhibitors:
    • Antimetabolites: Cytarabine (DNA Pol), Methotrexate (DHFR), 5-FU (Thymidylate Synthase).
    • Topoisomerase Inhibitors:
      • Type I: Irinotecan, Topotecan.
      • Type II: Etoposide, Doxorubicin.

⭐ Quinolone antibiotics (e.g., Ciprofloxacin) target bacterial DNA gyrase (Type II topoisomerase).

High‑Yield Points - ⚡ Biggest Takeaways

  • DNA replication: semiconservative, bidirectional, S phase. Key enzymes: Pol III/δ/ε, helicase, ligase.
  • Okazaki fragments form on the lagging strand, joined by DNA ligase.
  • Telomerase (reverse transcriptase) maintains eukaryotic telomere length, preventing degradation.
  • Mismatch Repair (MMR) corrects post-replication errors; defects cause Lynch syndrome.
  • Nucleotide Excision Repair (NER) removes UV-induced pyrimidine dimers; defects cause Xeroderma Pigmentosum.
  • Base Excision Repair (BER) corrects single damaged bases like deamination.

Practice Questions: DNA Replication and Repair Mechanisms

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Which of the following is true about ataxia telangiectasia?

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Flashcards: DNA Replication and Repair Mechanisms

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In Nonhomologous End Joining (NHEJ), dsDNA breaks are recognized by _____, recruiting the MRN complex which then recruits the Artemis protein and various others to help repair the damage

TAP TO REVEAL ANSWER

In Nonhomologous End Joining (NHEJ), dsDNA breaks are recognized by _____, recruiting the MRN complex which then recruits the Artemis protein and various others to help repair the damage

ATM

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