DNA Structure and Replication

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DNA Structure - Helix Blueprint

  • DNA: Deoxyribonucleotide polymer; genetic blueprint.
  • Components: Deoxyribose, phosphate, N-bases (A,T,G,C).
    • Purines: Adenine (A), Guanine (G) (📌 Pure As Gold).
    • Pyrimidines: Cytosine (C), Thymine (T) (📌 CUT Py).
  • Nucleoside: Base + Sugar. Nucleotide: Base + Sugar + Phosphate.
  • 5'-3' Phosphodiester bonds form backbone.
  • Watson-Crick B-DNA: Right-handed double helix.
    • Antiparallel strands.
    • Complementary base pairing (Chargaff's rules):
      • A=T (2 H-bonds)
      • G≡C (3 H-bonds)
      • Purines (A+G) = Pyrimidines (T+C)
  • Dimensions (B-DNA):
    • ~10.5 base pairs/turn.
    • Pitch: 3.4 nm/turn.
    • Diameter: 2 nm.
  • Major & minor grooves: Sites for protein binding. B-DNA double helix with major and minor grooves

⭐ B-DNA: right-handed helix; 10.5 bp/turn, pitch 3.4 nm. Z-DNA: left-handed.

DNA Packaging - Chromatin Coils

  • DNA (~2m) compacts into nucleus (~10µm) through hierarchical coiling.
  • Nucleosome ("beads-on-a-string"): Fundamental unit.
    • DNA (146 bp) wraps ~1.65 times around a histone octamer (2 each of H2A, H2B, H3, H4).
    • H1 histone (linker histone) binds to linker DNA and the nucleosome, aiding compaction.
    • Forms a 10-11 nm fiber.
  • Solenoid:
    • Helical coiling of 6 nucleosomes per turn.
    • Forms a 30 nm fiber.
  • Higher-order packaging: 30 nm fiber forms loops (300 nm), then scaffolds (700 nm), ultimately condensing into a metaphase chromosome (1400 nm). DNA packaging from double helix to duplicated chromosome

⭐ Histones are rich in basic amino acids (Lysine and Arginine), giving them a net positive charge that facilitates binding to the negatively charged DNA backbone.

DNA Replication - Copy Machine

DNA replication is a semi-conservative process, ensuring each daughter DNA molecule has one parental and one newly synthesized strand. It occurs primarily during the S-phase of the cell cycle.

  • Key Steps & Enzymes:
    • Initiation:
      • Origin of Replication (ORI): Specific DNA sequences where replication begins.
      • Helicase: Unwinds DNA double helix at replication fork (ATP-dependent).
      • Single-Strand Binding Proteins (SSBPs): Stabilize unwound single-stranded DNA (ssDNA).
      • Topoisomerases: Relieve supercoiling ahead of fork (e.g., DNA gyrase in prokaryotes).
    • Elongation:
      • Primase: Synthesizes short RNA primers.
      • DNA Polymerase III: Main prokaryotic synthesizing enzyme (5'→3' activity), requires RNA primer.
        • Leading strand: Continuous synthesis towards the fork.
        • Lagging strand: Discontinuous synthesis (Okazaki fragments) away from the fork.
      • DNA Polymerase I: Prokaryotic; removes RNA primers (5'→3' exonuclease activity), fills gaps with DNA.
    • Termination:
      • DNA Ligase: Joins Okazaki fragments on lagging strand, seals nicks in the phosphodiester backbone.

DNA Replication Fork Diagram

⭐ DNA replication is typically bidirectional from the origin, creating two replication forks. Prokaryotes usually have a single ORI, while eukaryotes possess multiple ORIs to ensure timely replication of larger genomes.

Replication Fidelity & Telomeres - Proof & Protect

  • Replication Fidelity (Error Prevention):
    • DNA Pol III (prok.) & Pol δ/ε (euk.) 3'→5' exonuclease activity.
    • Mismatch Repair (MMR): Corrects residual errors. Proteins: MutS/L/H (prok.), MSH/MLH/PMS (euk.).
    • Achieves error rate of $1 \times 10^{-9}$ to $1 \times 10^{-10}$.
  • Telomeres (Chromosome End Caps):
    • Repetitive DNA (human: TTAGGG) at chromosome ends; prevent gene erosion.
    • Counteract "end replication problem" (incomplete lagging strand synthesis).
    • Telomere shortening during DNA replication
  • Telomerase (Maintains Telomeres):
    • Ribonucleoprotein enzyme (reverse transcriptase activity).
    • Contains RNA template (TERC) & catalytic subunit (TERT).
    • Adds telomeric repeats. High in germ, stem, cancer cells. Low in somatic cells (→ Hayflick limit).
    • ⭐ > Telomerase inhibition is a therapeutic target in cancer treatment.

High‑Yield Points - ⚡ Biggest Takeaways

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Practice Questions: DNA Structure and Replication

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Which of the following molecular interactions are found in the structure of DNA?

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Flashcards: DNA Structure and Replication

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DNA & RNA are made of _____ connected via a phosphodiester bond

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DNA & RNA are made of _____ connected via a phosphodiester bond

nucleotide monophosphates

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