DNA Structure and Replication
Nucleotide Connection in Nucleic Acids
Nucleotides must be connected on the 3' side.
Nucleic acids are synthesized by adding nucleotides at the 3' end.
Polynucleotide Definition: A strand of nucleotides connected together.
Formation of Bonds
The bond that forms between nucleotides is called a phosphodiester bond.
Every new nucleotide must be added to the 3' end of the existing nucleotide within a polynucleotide strand.
Connection of nucleotides occurs by formation of phosphodiester bonds.
Structure of DNA
DNA consists of two polynucleotide strands.
Phosphates and sugars form the backbone (represented as blue ribbons).
Nitrogenous bases reside in the interior.
Types of Bonds Between DNA Strands
The strands of DNA are held together by hydrogen bonds between complementary bases.
Base Pairing Rules:
Guanine (G) pairs with Cytosine (C).
Adenine (A) pairs with Thymine (T).
Importance of base pairing rules:
Distance Maintenance: Essential for correct connection.
Hydrogen Bond Variation:
G and C form 3 hydrogen bonds.
A and T form 2 hydrogen bonds.
Cannot pair A with G or C with T due to hydrogen bond differences and distance issues.
Percent Composition of DNA Bases
If 20% of the bases are guanine (G), then cytosine (C) must also be 20% due to pairing rules.
Therefore:
%A + %T + %C + %G = 100%
A and T together must then account for 60%, so both A and T are 30% each.
Introduction to DNA Replication
DNA replication involves copying the DNA molecule.
Essential for cell division to ensure that new cells receive copies of DNA.
Known method of DNA replication is the semi-conservative hypothesis:
Each new DNA molecule contains one original strand and one newly synthesized strand.
Base pairing is vital as original strands serve as templates for new strands.
Prokaryotic vs. Eukaryotic DNA Replication
Prokaryotes have a circular chromosome, whereas eukaryotes have linear chromosomes.
Most replication details are similar between prokaryotes and eukaryotes.
Steps of DNA Replication
Separation of Strands:
DNA helix is unwound and separated at the replication fork.
Parent strands serve as templates.
DNA Polymerase:
Can only add nucleotides to the 3' end of the growing strand.
Builds new strands antiparallel to the template strands.
Leading and Lagging Strands
Leading Strand:
Built continuously towards the replication fork.
Lagging Strand:
Built in fragments (Okazaki fragments) because it is built away from the replication fork, requiring multiple RNA primers.
Enzymes in DNA Replication
Helicase:
Unwinds and separates DNA strands by breaking hydrogen bonds.
Single-Strand Binding Protein:
Stabilizes separated strands to prevent re-association.
Topoisomerase:
Relieves the tension of coiling ahead of the replication fork.
Primase:
Synthesizes short RNA primers to initiate DNA synthesis.
DNA Polymerase III:
Main enzyme that synthesizes new DNA strands.
DNA Ligase:
Joins Okazaki fragments on the lagging strand by forming phosphodiester bonds.
Final Steps and Completion of DNA Replication
RNA primers must be replaced with DNA nucleotides, carried out by DNA Polymerase I.
After replacing RNA primers, ligase connects any remaining fragments, completing DNA replication.
Result is two identical DNA molecules, each containing one original and one new strand.