Bio 1 Flashcards
Nucleic Acid
Macromolecule composed of nucleotide monomers.
DNA and RNA (mRNA, tRNA, rRNA, snRNA, etc.).
Structure: linear strand of nucleotides joined by phosphodiester bonds.
May be single or double stranded.
Nucleotide
Nitrogenous base + sugar + 1-3 phosphates.
Nucleic Acid “backbone”: sugar and phosphate group of each nucleotide linked by phosphodiester bonds.
Glycosidic bond between sugar and the base; Hydrogen bonds between the bases if double stranded.
DNA and RNA differ in the 2’ carbon of the sugar; RNA (ribose) has an OH; DNA (deoxyribose) has H.
Base linked to the 1’C of sugar, phosphate linked to 5’C.
Nitrogenous Bases
Purines: Adenine/Guanine (double ring).
Pyrimidines: Cytosine, Uracil, Thymine (single ring).
U and T differ in a methyl group.
Base Pairing
A always pairs with T (with U in RNA -> 2H bonds).
C always pairs with G
-> 3H bonds (stronger base pair)
DNA Replication: Overview
Overall: one DNA molecule replicated -> 2 identical copies .
- Occurs during the S phase (synthesis phase) of the cell cycle.
- Cell division requires DNA replication to occur.
- Semi-Conservative nature
- The synthesis of the new “daughter strand” occurs on top of the “parent strand” through the addition of free floating nucleotides.
DNA replication requires a template (the parent DNA strand), as well as a primer to jumpstart the synthesis (RNA primer).
DNA is built in the 5’ -> 3’ direction of both daughter strands.
Semi-Conservative nature of DNA replication
- When the mechanism of DNA replication was first discovered, there were 3 different methods proposed; semi-conservative was found to be correct.
- The strands of the original DNA molecule separate and serve as templates for the synthesis of new, complimentary strand.
Enzymes involved in DNA Replication
DNA Helicase
Topoisomerase
DNA Gyrase
Single Strand Binding (SSB) Proteins
Primase
DNA Polymerase
Ligase
Step 1 of DNA Replication
Beginning at a specific region of the DNA (the ORI), helicase binds to the DNA and begins to unwind it by breaking H-bonds between bases -> forms a replication bubble with replication forks on either end.
-ORI found by other helper proteins.
Step 2 of DNA Replication
Topoisomerase relaxes supercoils that occur upstream of the replication bubble due to to the unwinding of the strands.
-Cuts DNA and unwinds the excess coils.
Step 3 of DNA Replication
SSBPs stabilize the newly separated parent strands, making sure they don’t reanneal.
Step 4 of DNA Replication
Primase lays down a few RNA bases that are complimentary to the parent strand and serve as a primer, from which DNA polymerase can build off of.
- DNA Pol can only extend, can’t initiate
- These RNA primers must later be removed and replaced with DNA.
Step 5 of DNA Replication
DNA Polymerase elongates from the primer, adding nucleotides one by one and using the parent strand as a template.
- Free floating dNTPs are added (triphosphate) and the release of two phosphate groups from the molecule provides the energy for this reaction.
- 3’ hydroxyl group on the last nucleotide of the daughter strand performs nucleophillic attack onto phosphate (at 5’ end) of the incoming nucleotide.
Step 6 of DNA Replication
At each replication fork, there is a leading strand and a lagging strand of the daughter molecules, due to the requirement that the new strands are built in 5’ -> 3’ direction of elongation.
Step 7 of DNA Replication
Leading strand is synthesized in the same direction as the extending replication fork and is a single continuous strand.
Step 8 of DNA Replication
Lagging strand is not continuous and made up of multiple Okazaki fragments that are soon after joined together by DNA ligase.
Step 9 of DNA Replication
Synthesis is bidirectional because it extends in both directions from the replication bubble.
Step 10 of DNA Replication
RNA primers are eventually replaced with DNA using DNA Polymerase
Eukaryotes
- Process occurs in Nucleus
- Many ORI per DNA molecule (linear DNA) -> multiple replication bubbles at once; ultimately the bubbles meet each other and the newly formed DNA molecules separate.
- Many simultaneous replication bubbles enables fast replication
- 5 DNA polymerases
- Only eukaryotes have telomeres.
Prokaryotes
-The process occurs in the Cytosol
-One ORI per DNA molecule (circular DNA) -> theta replication
-3 DNA polymerases
DNA Pol III: main DNA Pol that extends the daughter strands.
DNA Pol I: slower than Pol III; exonuclease activity enables it to remove RNA primer and replace with DNA; also plays a role in DNA error repair