UNIT 2: DNA Damage and DNA Repair

Exam Focus: Distinguish between the types of damage (e.g., base alteration vs. strand break). Focus on the core excision repair pathways: **Base Excision Repair (BER)** and **Nucleotide Excision Repair (NER)**, outlining their step-by-step mechanism.

Table of Contents

  1. DNA Damage
  2. DNA Repair

1. DNA Damage

DNA damage refers to changes in the DNA structure, often caused by natural cellular processes or external mutagens.

Deamination

The loss of an amino group from a base. For example, deamination of cytosine produces uracil, which is highly mutagenic if not repaired before replication.

Depurination

The cleavage of the glycosidic bond between the base and the sugar, resulting in the loss of a purine base (A or G) from the DNA backbone.

Altered bases

Chemical modification of a base, often caused by alkylating agents, radiation, or oxidizing agents, which can lead to mispairing during replication.

Single strand breaks

Disruption in the sugar-phosphate backbone of one DNA strand. These are generally easy to repair using the intact complementary strand as a template.

Double-strand breaks (DSBs)

Breaks in the sugar-phosphate backbone of both DNA strands, a highly dangerous form of damage that can lead to chromosomal rearrangements or cell death.

Cross-linking

Covalent bonding between bases within the same strand (intrastrand) or between opposing strands (interstrand), preventing strand separation for replication or transcription. A common example is the formation of **thymine dimers** caused by UV radiation.

DNA methylaton

The addition of a methyl group, usually to cytosine bases in CpG dinucleotides. While a key process in gene regulation, it can lead to spontaneous deamination to thymine, causing a G-T mismatch.

2. DNA Repair

Cells possess multiple enzymatic pathways to detect and repair damaged DNA, ensuring genomic stability.

Photoreactivation

A direct repair mechanism (mostly in prokaryotes) that reverses **thymine dimers** caused by UV light. The enzyme **photolyase** uses energy from visible light to break the cross-links.

Base excision repair (BER)

Repairs minor damage involving a single, non-helix-distorting base lesion (e.g., altered bases like uracil from deamination).

  1. **Glycosylase** recognizes and removes the damaged base, leaving an **AP (apurinic/apyrimidinic) site**.
  2. **AP Endonuclease** cleaves the phosphodiester backbone at the AP site.
  3. **DNA Polymerase** inserts the correct nucleotide.
  4. **DNA Ligase** seals the nick.

Nucleotide excision repair (NER)

Repairs bulky, helix-distorting lesions (e.g., thymine dimers, chemically modified bases).

  1. Damage is recognized by specialized proteins.
  2. A segment of the damaged strand, containing the lesion, is excised (removed).
  3. **DNA Polymerase** uses the complementary strand as a template to synthesize the replacement patch.
  4. **DNA Ligase** seals the gap.
[Image of Nucleotide Excision Repair mechanism]

Mismatch repair

Corrects errors (mismatched bases or small loops) incorporated during DNA replication that were missed by the polymerase proofreading function. It identifies and excises the mismatched segment from the newly synthesized strand using methylation markers on the parental strand for strand discrimination.

Translational synthesis (Translesion Synthesis)

A **bypass mechanism** used when the replication fork encounters major damage that cannot be excised quickly. A special DNA polymerase (translesion polymerase) bypasses the lesion, often introducing errors, but allowing replication to continue (error-prone).

Recombinational repair

Uses homologous DNA sequences to accurately fill gaps or repair breaks in the DNA strand.

Homologous recombination (HR)

A highly accurate mechanism for repairing **Double-Strand Breaks (DSBs)** by using the sister chromatid or homologous chromosome as a perfect template.

Non-homologous end-joining (NHEJ)

An error-prone pathway for repairing DSBs by simply trimming the broken ends and ligating them together. It does not require a homologous template and is prone to loss of genetic material.