Notes on DNA Structure, Packaging, and Replication
Acknowledgement
- Acknowledge the Traditional Owners of the land: Wurundjeri People of the Kulin Nation.
- Respect to Elders: past, present, and emerging.
- Recognition of connection to Wurundjeri Country and commitment to respect Indigenous heritage.
Topic Learning Objectives
- Basic Properties of DNA:
- Understand nucleotides, complementary base pairing, double-strand helix structure.
- DNA Packaging and Regulation:
- Explore how DNA is organized within the cell.
- DNA Replication Process:
- Describe the enzymes involved in DNA replication and their functions.
- Leading vs Lagging Strand Replication:
- Compare and contrast replication on leading and lagging strands.
Introduction to DNA, Chromosomes, and Genome
- DNA Structure:
- Eukaryotic DNA consists of two complementary strands of nucleotides.
- Nucleotides include:
- Guanine (G)
- Adenine (A)
- Cytosine (C)
- Thymine (T)
- Covalent bonds form the sugar-phosphate backbone with orientation indicated by 5’ and 3’ ends.
Genome
- Definition:
- Complete set of genes or genetic material in a cell or organism.
- Usage of DNA:
- Chromosomes carry genes, which are segments of DNA that encode for proteins.
Chromosome Organization
- DNA Hybridization:
- Visualizes chromosomes and their structure as per karyotypes.
- Coding vs Non-Coding DNA:
- Only 1% of DNA codes for proteins.
- Exons: coding regions.
- Introns: noncoding regions.
- Regulatory DNA sequences control gene expression and protein production.
Cell Cycle and DNA Replication
- Cell Cycle Phases:
- Interphase: Genes expressed, chromosomes replicated.
- M-phase: Chromosomes condense and divide.
- Specialized DNA Regions:
- Replication origin, centromere, telomere.
DNA Packaging
- Nucleosome Formation:
- Nucleosomes compact DNA extensively (e.g., Chromosome 22 contains 48 million nucleotide pairs).
- Chromatin Types:
- Euchromatic: less condensed (active).
- Heterochromatic: more condensed (inactive).
DNA Replication Mechanism
- Enzymes in Replication:
- Helicase: Unwinds the double-stranded DNA.
- SSB Proteins: Stabilize separated strands.
- DNA Polymerase: Synthesizes new DNA strands in the 5’ to 3’ direction.
- Replication Fork:
- Multiple enzymes operate at replication forks to synthesize daughter strands.
- Lagging strand synthesized in fragments (Okazaki fragments).
Leading vs Lagging Strand
- Leading Strand:
- Synthesized continuously in the 5’ to 3’ direction.
- Lagging Strand:
- Synthesized as Okazaki fragments, requires RNA primer (made by DNA primase) to start synthesis.
- Fragments joined by DNA ligase post-synthesis.
DNA Polymerase Functions
- Proofreading Ability:
- Exonucleolytic proofreading increases fidelity during DNA replication (errors decreased to 1 in 10 billion).
Telomere Maintenance
- Function of Telomeres:
- Composed of repeated sequences (e.g., GGGTTA).
- Telomerase replenishes telomeric sequences during cell division, counteracting the potential shortness from incomplete replication at lagging strands.
- Reverse Transcriptase Activity:
- Uses RNA template to extend the lagging strand, allowing for complete DNA synthesis.