DNA Replication and Chromosomes Flashcards
What did the 3-dimensional crystal structure *proposed by Watson ad Crick suggested?
How DNA could be replicated to maintain the genetic information.
What could each strand of DNA do?
Separated.
Used as the template for second strand production.
What should DNA duplex be before any synthesis begins?
Unwound.
Melted apart.
With what do enzymes need to deal?
With the 5’-3’ orientation of each of the DNA strands.
In what do the 3 replication models of DNA differ?
In terms of how much old and new DNA daughter cells contain.
What were the thoughts of DNA helix unwound and replication?
Unclear if it would happen along the entire bacterial chromosome in one go.
If it had to be done in multiple smaller sections.
How could the 3 competing models of DNA replication be tested based on Meselson and Stahl?
By labelling Escherichia coli DNA during replication with nitrogen isotopes.
Where did incorporation of 14N resulted?
In light DNA.
Where did use of 15N resulted?
In heavier DNA.
How could light and heavy DNA be separated?
By ultra-centrifugation.
Where is DNA centrifuged?
In a CsCl gradient.
What happens to DNA when it is centrifuged in a CsCl gradient?
It moves towards neutral buoyancy point.
Which are the 3 models of DNA strands?
Conservative.
Semi-conservative.
Dispersive.
What happened in Meselson-Stahl experiment?
Several generations of bacteria went through it.
Chromosomal DNA became increasingly ‘light’.
When is replicating/synthesising DNA simple?
When a single template strand is provided.
What happens while the double-stranded DNA is replicated?
One strand is in the opposite direction to the other.
The 2 DNA Polymerases collide.
How do the cells avoid the 2 DNA Polymerases from colliding?
By flexing one of the 2 strands around.
What happens to the 2 DNA Polymerases when the cells flex one of them around?
They move in the same direction as a single ‘DNA synthesis/replication machine’.
For what is a replication complex/machine responsible?
DNA replication in vivo.
What does the replication complex/machine generate?
Copies of whole chromosomes in each cell cycle.
What does DNA Polymerase synthesize?
New DNA.
What does DNA Polymerase catalyse?
Formation of phosphodiester bond between 3’-OH of DNA strand synthesized and incoming 5’-triphosphate (dNTP).
What does the parental DNA strand provide?
The template for base pairing.
From where does the energy for the parental DNA providing the template come from?
The removal of pyrophosphate from incoming dNTP.
What must be free for the parental DNA providing the template?
3’OH group.
In which direction does polymerisation always occur?
In 5’–>3’.
What does DNA Polymerase test?
Each new dNTP.
What does DNA Polymerase required?
The correct base pairing before moving to the next base.
What is the direction of DNA Polymerase activity?
3’-5’.
How is the activity of DNA Polymerase named?
Exonuclease.
What does the exonuclease activity 3’-5’ of DNA Polymerase allow it to do?
Remove a mistake.
Replace the wrong nucleotide with the correct one.
What do RNA primers initiate?
DNA synthesis.
What does DNA Polymerase need to begin polymerisation?
A 3’-OH group.
What can DNA Primase start, without requiring starting point?
An RNA-based polynucleotide chain.
What does DNA Primase provide?
A ‘primer’ for DNA Polymerase.
What does primases uses instead of deoxyribonucleotides?
Ribonucleotides.
For how long does Primase function?
For only very short time.
Why does primase function for only a very short time?
Because RNA is unstable.
NTPs are limited.
How long is the primer?
Around 10 nucleotides long.
What does primase incorporate?
dNTPs.
Why does primase incorporates dNTPs?
To provide a stronger synthesis scaffold.
By what can DNA Polymerase synthesize DNA?
From the leading strand.
What is the characteristic of the other/lagging strand of DNA?
It is in the ‘wrong’ orientation than the leading strand.
How is the problem of wrong orientation of lagging strand synthesis fixed?
By breaking down the synthesis into small sections.
Which are the small sections lagging strand break down to?
Okazaki fragments.
How are Okazaki fragments linked together?
By DNA ligase.
What do helicases unwind?
The parental double helix.
What do single-strand binding proteins stabilise?
The unwound parental DNA.
From what are the 2 parental strands of DNA replicated?
By 2 DNA Polymerases.
Where are the 2 parental strands of DNA while they are replicated by 2 DNA Polymerases?
In a single replication machine.
Where does e single replication machine travel during DNA replication by DNA Polymerases?
Along the parental DNA duplex.
What does the single replication machine do to the parental DNA duplex?
Unwinding.
Melting as goes.