DNA REPLICATION Flashcards

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1
Q

DNA STRUCTURE

A

Nucleic acids: Biopolymers built from smaller molecules (monomers) connected covalently.

DNA structure: Double helix with a sugar-phosphate backbone and nitrogenous bases (A, G, C, T).

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2
Q

DNA replication overview

A

Goal: To ensure each daughter cell receives one copy of DNA.

Semi-conservative replication: Each new DNA molecule has one old and one new strand.

Other theories: Conservative and dispersive models (disproven).

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3
Q

challenges

A
  1. Replicate all bases of long DNA accurately.
  2. Minimize replication errors.
  3. Overcome mechanical stress during unwinding and encounters with transcribed genes.
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4
Q

DNA polymerase and proofreading

A

DNA polymerase: Efficient at proofreading, with 1 error per 10^7 bases.

Exonuclease activity: Removes mismatches during replication to ensure accuracy.

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5
Q

basic nucleotide structure

A

DNA: A, G, C, T bases with deoxyribose sugar.

RNA: A, G, C, U bases with ribose sugar.

Sugar-phosphate backbone supports 5’ to 3’ polymerization.

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6
Q

bacterial genome replication

A

Circular DNA: Replication begins at a single origin (oriC) and proceeds bidirectionally.

Replication is not synchronized with cell division.

Plasmids: May have variable copy numbers.

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7
Q

initiation of DNA replication

A

oriC: A-T rich region where DNA melts due to weaker bonds.

DnaA protein binds to the origin, causing strand separation and allowing helicase attachment.

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8
Q

supercooling and topoisomerase

A

As DNA unwinds, supercoiling occurs.

Topoisomerases: Remove supercoiling to prevent replication machinery from stalling.

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9
Q

leading and lagging strands

A

Leading strand: Continuous synthesis in the 5’ to 3’ direction.

Lagging strand: Synthesized in short fragments (Okazaki fragments) joined by DNA ligase.

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10
Q

key replication enzymes

A

DNA polymerase III: Extends the DNA strand but cannot start it.

Primase: Synthesizes an RNA primer for DNA pol III to extend.

Helicase: Unwinds the DNA strands.

DNA polymerase I: Removes the RNA primer and fills gaps on the lagging strand.

DNA ligase: Joins Okazaki fragments.

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11
Q

sugar structure in nucleotides

A

C1: Binds the base.

C2: No OH in DNA; OH present in RNA.

C3 and C5: Interact with phosphate groups for polymerization.

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12
Q

double stranded DNA rules

A

Anti-parallel strands.

Base-pairing: A with T, G with C.

Nucleotides added to the 3’ end using energy from phosphate bond hydrolysis.

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13
Q

polarity in DNA

A

5’ end: Phosphate group.

3’ end: Hydroxyl group.

Results in directionality from 5’ to 3’ in synthesis.

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14
Q

eukaryotic genome replication

A

Requires multiple origins due to larger, linear chromosomes.

Euchromatin origins fire first (early origins).

Heterochromatin origins fire later (late origins).

Replication occurs during the S-phase of the cell cycle.

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15
Q

replicons

A

DNA region replication from a single origin

Bacteria: Each circular DNA is a single replicon.

Eukaryotes: Have multiple replicons, and not all origins fire in every cycle. Some are dormant.

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16
Q

replication forks

A

Prokaryotes: Helicase moves along the lagging strand template.

Eukaryotes: Helicase moves along the leading strand template.

Pol ε: Synthesizes the leading strand in eukaryotes.
Pol δ: Synthesizes the lagging strand in eukaryotes.

17
Q

protein phosphorylation in replication

A

Protein phosphorylation regulates origin firing during the S-phase.

Ensures each origin fires only once per cell cycle.

18
Q

end replication problem

A

Each replication cycle leaves unreplicated 50-200 bp at the 3’ end.

Telomeres: Non-coding, repetitive DNA sequences (e.g., TTAGGG) protect chromosome ends.

19
Q

Telomeres and Aging

A

Telomeres shorten with each replication cycle, limiting cell proliferation.

Short telomeres protect against cancer, but in cancer cells, telomere length is stabilized.

20
Q

Telomerase

A

Telomerase: An enzyme that extends telomeres, compensating for DNA loss during replication.

Functions as a reverse transcriptase.

21
Q

types of circular DNA

A

ssDNA or dsDNA: No DNA ends.

Circular nicked: Backbone broken, but all bases paired.

Circular gapped: Contains unpaired bases.

22
Q

types of linear DNA

A

Linear dsDNA or ssDNA: No gaps in structure.

Linear nicked: Backbone is broken.

Linear gapped: Contains unpaired bases.

23
Q

types of ends in liner DNA

A

Blunt ends: No overhangs.

3’ overhang: Extra nucleotides on the 3’ end.

5’ overhang: Extra nucleotides on the 5’ end.

24
Q

branched DNA structures

A

3-way junctions: Seen in replication forks.

4-way junctions: Recombination or repair intermediates.

25
Q

DNA end representation

A

3’ end: Always marked with an arrowhead.

5’ end: No arrowhead.

26
Q

key enzymes in DNA PROCESS

A

Helicase: Unwinds DNA strands.

Nuclease: Cleaves the sugar-phosphate backbone.

Polymerase: Synthesizes nucleic acids.

Clumps: Assist enzymes, not enzymes themselves.

Ligase: Joins nucleotides together.

27
Q

helices types

A

Hexameric helicases: DNAb (prokaryotes), MCM (eukaryotes).

Monomeric helicases: Involved in DNA repair and replication assistance.

28
Q

helicase energy source and directionality

A

Energy source: ATP hydrolysis for movement and unwinding.

DNAb direction: 5’ to 3’ (prokaryotes).

MCM direction: 3’ to 5’ (eukaryotes).

29
Q

nuclease overview

A

Break the sugar-phosphate backbone at C3 or C5.

Can be specific to ssDNA or dsDNA.

30
Q

types of nuclease

A

5’ to 3’ exonucleases: Cleaves one nucleotide at a time, cannot cleave circular DNA.

Endonucleases: Cleaves within DNA without needing ends, can cleave circular DNA.

3’ to 5’ exonucleases: Cannot cleave circular DNA.

31
Q

helices nuclease team

A

Helicase unwinds dsDNA, creating a substrate for ssDNA nucleases.

Commonly seen in DNA repair and generation of 3’ overhangs.

32
Q

DNA polymerase directionality

A

All polymerases synthesize DNA in the 5’ to 3’ direction by adding nucleotides to the 3’ end.

Cannot extend the 5’ end in the cell.

33
Q

DNA polymerase proofreading

A

Uses 3’ to 5’ exonuclease activity to remove incorrect, non-complementary nucleotides.

34
Q

DNA polymerase processivity

A

Processivity: The ability to extend a DNA strand without dissociating.

Higher processivity in vivo compared to in vitro.

35
Q

eukaryotic clump - PCNA

A

PCNA: A structural protein that forms a closed ring, stabilizing replication and repair enzymes on DNA.

Clump loader: An enzyme that opens the PCNA ring using ATP.

clumped loader binds to DNA and clump, open clump and loads onto DNA

clump loader releases clump and dissociates from DNA