L7: DNA Replication Flashcards

Learning Objectives: • Outline the mechanism of DNA replication in prokaryotes. • Outline the importance and processes of proofreading and error correction by DNA polymerases. Outline key differences between the mechanisms of DNA replication in prokaryotes and eukaryotes.

1
Q

Why is DNA replication important?

A

New cells are created by mitosis, which required DNA replication

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2
Q

What happens in DNA replication?

A

Identical copy of the parent DNA is used as a template for a new strand

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3
Q

Define semi-conservative replication

A

2 copies of the original DNA is produced, consisting of only half of the material

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4
Q

When is the replication fork created?

A

DNA helicase unwinds the double helix structure

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5
Q

Initiation of DNA replication in prokaryotes

A

1) Initiation proteins bind to replication origin
2) Separate strands of DNA
3) Two ‘replication forks’ are formed

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6
Q

Which directions do replication forks move?

A

In opposite directions

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7
Q

Which bonds are nucleotides joined by?

A

Phosphodiester Bonds

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8
Q

Structure of deoxynucleoside triphosphates (dNTPs)

A

3 phosphate groups attached to the 5’ carbon of deoxyribose

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9
Q

Role of DNA polymerase III

A

Synthesis the bulk of DNA in E.coli

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10
Q

What is a holoenzyme?

A

Composed of multiple subunits with different functions

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11
Q

What does DNA pol III require for DNA synthesis?

A

RNA primer

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12
Q

How is RNA primer created ?

A

Using enzyme PRIMASE

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13
Q

Which direction does DNA pol III synthesis new DNA in?

A

5’ to 3’ direction

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14
Q

What is the leading strand?

A

Replicates continuously

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15
Q

What is the lagging strand?

A

Replicates to form short fragments

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16
Q

Polarity problem in DNA replication

A

DNA double helix have opposite polarity but all DNA polymerase replicate in the same direction

17
Q

What are Okazaki fragments joined by?

A

DNA ligase

18
Q

Role of ligase in DNA replication

A

Joins Okazaki fragments into continuous DNA strands

19
Q

Function of enzyme topoisomerase

A

Preventing DNA from getting tangled
- Relieving pressure supercoiling ahead of replication fork

20
Q

Role of Type I topoisomerases

A

Cut 1 DNA strand

21
Q

Role of Type II topoisomerases

A

Cut both DNA strands, pass coil through gaps (ATP) & reseal the strands

22
Q

Role of helicase in DNA replication

A

Separates DNA strands

23
Q

Role of single-stranded binding protein in DNA replication

A

Prevents DNA strands from re-annealing

24
Q

Role of primase in DNA replication

A

Synthesise RNA primers

25
Role of DNA polymerase III
Synthesise bulk of DNA
26
Role of sliding clamp in DNA replication
Keeps DNA pol III firmly attached to DNA
27
What enzymes act as proofreaders during DNA polymerisation in DNA replication?
Exonucleases
28
How does the exonuclease activity proofread?
Remove last added nucleotide via exonuclease activity, then insert the correct nucleotide
29
Which DNA polymerase has exonuclease activity?
DNA polymerase I and III
30
What is a mismatch repair?
Errors missed by proofreading activity, corrected by repair machinery
31
How does the mismatch repair work?
1) Identify incorrect base pairs 2) Remove DNA in new strand 3) Synthesise replacement DNA
32
What is DNA replication in eukaryotic cells?
Part of the 'cell cycle'
33
Where is DNA replication in eukaryotic cells
Nucleus
34
Where is DNA replication in prokaryotic cells?
Cytoplasm
35
Role of aminoacyl -tRNA synthetase in protein synthesis
Joining the amino acids to tRNA
36
Role of peptidyl transferase in protein synthesis
Formation of the peptide bond