Lecture 6 - DNA Replication Flashcards

1
Q

What are nucleic acids made of?

A

Nucleic acids (DNA and RNA) are biopolymers made of nucleotide monomers, which consist of a sugar, phosphate group, and nitrogenous base.

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2
Q

What sugars are found in nucleotides?

A

DNA contains deoxyribose, while RNA contains ribose.

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3
Q

How are nucleotides linked in DNA and RNA?

A

Nucleotides are linked through phosphodiester bonds between the 5’ carbon of one sugar and the 3’ carbon of the next.

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4
Q

What gives DNA and RNA strands polarity?

A

The 5’ end (with a free phosphate) and the 3’ end (with a free hydroxyl group) create polarity.

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5
Q

What is the base-pairing rule in DNA?

A

Adenine pairs with thymine (A-T) and cytosine pairs with guanine (C-G) via hydrogen bonds.

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6
Q

What is the role of DNA polymerase?

A

DNA polymerase synthesises DNA by adding nucleotides to the 3’ end of a growing strand, using a DNA template.

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7
Q

Where does the energy for DNA synthesis come from?

A

Energy is provided by the high-energy phosphate bonds in deoxyribonucleotide triphosphates (dNTPs).

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8
Q

What is a key limitation of DNA polymerase?

A

DNA polymerase cannot initiate synthesis on its own; it requires a primer with a free 3’ end.

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9
Q

What are helicases?

A

Helicases are enzymes that unwind DNA strands by breaking hydrogen bonds between base pairs.

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10
Q

What are the roles of topoisomerases during replication?

A

Topoisomerases relieve supercoiling stress ahead of the replication fork.

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11
Q

How is DNA replication initiated in prokaryotes?

A

Prokaryotic replication begins at a single origin of replication, such as oriC in E. coli, which includes an AT-rich region and DNA-binding sites for initiator proteins.

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12
Q

Why do eukaryotes have multiple origins of replication?

A

Eukaryotic genomes are much larger and organised into linear chromosomes, necessitating multiple origins for efficient replication.

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13
Q

What enzymes synthesise DNA on the leading and lagging strands in eukaryotes?

A

DNA polymerase epsilon synthesises the leading strand, and DNA polymerase delta synthesises the lagging strand.

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14
Q

What is the role of telomerase in eukaryotes?

A

Telomerase extends telomeres to solve the end-replication problem, preventing progressive chromosome shortening.

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15
Q

How does DNA polymerase ensure fidelity during replication?

A

DNA polymerase has proofreading activity, allowing it to remove mismatched bases using exonuclease activity.

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16
Q

What is the significance of Okazaki fragments?

A

Okazaki fragments are short DNA segments synthesised on the lagging strand, later joined by DNA ligase.

17
Q

How does replication differ on the leading and lagging strands?

A

The leading strand is synthesised continuously, while the lagging strand is synthesised discontinuously in Okazaki fragments.

18
Q

How is bacterial DNA replication unique?

A

Bacteria have circular chromosomes with a single origin of replication, enabling bidirectional synthesis.

19
Q

What is a replicon?

A

DNA molecule containing an origin of replication region essential for initating replication. can replictae with its own initiator and termination sequence.

20
Q

What are telomeres?

A

Telomeres are repetitive, non-coding DNA sequences at chromosome ends, protecting genetic material from degradation.

21
Q

How does telomerase function?

A

Telomerase extends telomeres by adding repetitive sequences using its RNA template, preventing chromosome shortening.

22
Q

What does “semi-conservative replication” mean?

A

Each daughter DNA molecule contains one parental strand and one newly synthesised strand.

23
Q

Why is DNA replication in eukaryotes tightly regulated?

A

Replication is regulated to ensure that each part of the genome is replicated once per cell cycle, avoiding errors or duplications.

24
Q

How do bacteria prevent over-replication?

A

Bacteria regulate DNA binding proteins (like DnaA) at the origin of replication, ensuring initiation occurs only once per cycle.

25
Q

How is helicase loaded onto DNA?

A

Helicase is loaded at the replication origin by initiator proteins, which open up the AT-rich region to begin unwinding.

26
Q

What is the function of DNA ligase?

A

DNA ligase seals nicks in the sugar-phosphate backbone, joining Okazaki fragments on the lagging strand.

27
Q

What causes supercoiling during DNA replication?

A

The unwinding of DNA strands by helicase generates supercoils ahead of the replication fork, relieved by topoisomerases.

28
Q

What are the roles of clamping proteins in DNA replication?

A

Clamping proteins increase the processivity of DNA polymerase, helping it stay attached to the DNA strand during synthesis.

29
Q

What is the role of the clamping loader in DNA replication?

A

Clamping loaders load and unload the sliding clamps onto DNA, ensuring proper functioning of DNA polymerase.

30
Q

What happens to bacterial DNA replication if DnaA protein binding is impaired?

A

Replication initiation at the oriC region would fail, as DnaA binding is essential for melting the AT-rich region and helicase loading.

31
Q

Why is telomere shortening a problem for linear chromosomes?

A

During replication, the RNA primers at the ends of chromosomes cannot be fully replaced by DNA, leading to progressive loss of genetic material.

32
Q

How does the bacterial replication fork differ from the eukaryotic fork?

A

In bacteria, the helicase moves on the lagging strand, while in eukaryotes, it moves on the leading strand.

33
Q

Why do eukaryotes require sequential firing of replication origins?

A

Sequential firing ensures efficient replication of large genomes while coordinating replication timing with the cell cycle.

34
Q

What is the role of primase in DNA replication?

A

Primase synthesises short RNA primers needed for DNA polymerase to initiate synthesis.

35
Q

How do eukaryotic cells coordinate lagging strand synthesis?

A

DNA polymerase delta removes RNA primers and synthesises the full Okazaki fragment, with ligase sealing nicks in the backbone.