7.1 DNA Structure and Replication (HL) Flashcards
Who understood that it was DNA and not protein that is the genetic material of a cell?
Alfred Hershey and Martha Chase
How did we understand that DNA is the genetic material of cells?
One batch of bacteriophages were grown in a radiolabelled sulphur (present in proteins) medium and another was grown in radioactive phosphorous (present in DNA)
They then infected E. coli and found only the phosphorous was passed on by gene transmission and not the sulphur
How did Rosalind Franklin and Maurice Wilkins investigate the structure of DNA?
Using X-ray diffraction
Describe the process of X ray diffraction
DNA is purified and the fibres are stretched in a thin glass tube
This is then targeted with an X ray beam which is diffracted when it hits an atom
This creates a scatter pattern
What inferences can be made from DNA scatter patterns?
DNA is double stranded
Nitrogenous bases are closely packed together on the inside and phosphates form an outer backbone
The DNA twists at regular intervals to form a double helix
How often does the DNA molecule twist?
Every 34 Angstrom
Who proposed the current structure of the DNA model?
James Watson and Francis Crick
Which molecules in DNA are purine?
Adenine
Guanine
What molecules are pyrimidines?
Cytosine
Thymine
How do adenine and thymine bond?
2 H bonds
How do guanine and cytosine bond?
3 H bonds
What is the function of helicase?
Unwinds and separated the double stranded DNA (replication)
How does helicase complete it’s function?
Breaking hydrogen bonds between base pairs
What is an origin of replication?
A specific region which is replicated
What is a replication fork?
The name given to the two strands running in antiparallel direction after being unwound by Helicase
What is the function of DNA Gyrase?
Reduces the torsional strain from unwinding (from helicase)
How does DNA Gyrase complete it’s function?
Relaxing the supercoils (via negative supercoiling)
What are SSB proteins?
Single stranded binding proteins
What is the function of SSB proteins?
Prevent re-annealing
Prevent the single stranded DNA from being digested by nucleases
When do SSB proteins dislodge from the DNA?
When a complimentary strand is synthesised onto the single strand
What is the function of DNA Primase?
Generates a short RNA primer on each template strand
What is needed before any base pairs can be added to a new nucleotide strand?
Primers
How long are most RNA primers?
(10 to 15 nucleotides)
What is the function of an RNA primer?
It is an initiation point for DNA polymerase III
What is the function of DNA polymerase III?
To synthesise a new complementary strand of DNA onto the original parent strand
On what end of the primer does DNA polymerase III attach?
3’
In which direction does DNA polymerase synthesise?
5’-3’
How does DNA pol III move in two directions art once?
On the leading fork DNA pol II moves towards the replication fork
On the lagging strand DNA pol III moves away from the replication fork and synthesises in pieces
What is the function of DNA polymerase I?
Removing the extra RNA primers that have to be added to create the Okazaki fragments on the lagging strand
What is the function of DNA ligase?
Joins the Okazaki fragments together to form a continuous strand
How are Okazaki fragments joined?
Covalently bonded joining the sugar-phosphate backbones together with phosphodiester bonds
In replication, which strand is the leading strand?
The one where DNA pol moves towards the fork and thus copies continuously
In replication, which strand is the lagging strand?
DNA pol moves away from the replication fork and copies discontinuously
What is a ddNTP?
A dideoxynucleotide does not have the 3’ hydroxyl group necessary to form phosphodiester bonds therefore ending replication as they cannot be elongated
How can ddNTP and PCR help us in sequencing?
PCR can be used to generate all the possible, terminating fragments for a particular base
These can then be ordered by length and thus the base sequences can be determines
What is the sanger method?
A method for determining the sequencing of DNA
What proportion of our DNA is non-coding?
More than 98%
Give examples of non-coding DNA
Satellite DNA Telomeres Introns Non-coding RNA genes Gene regulatory sequences
What is the function of satellite DNA?
Structural component of heterochromatin and centromeres - used in DNA profiling
What is the function of telomeres?
Regions of repetitive DNA at the end of the chromosome that protect against deterioration during replication
What is the function of introns?
Non-coding sequences within genes
How are introns removed?
By splicing before the formation of mRNA
What is the function of non-coding RNA genes?
Codes for RNA molecules which are not translated into protein
Give an example of a non-coding RNA gene
The genes for tRNA
What is the function of gene regulatory sequences?
Sequences that are involved in the process of transcription
Give examples of gene regulatory sequences
Promoters
Enhancers
Silencers
What are STRs?
Short tandem repeats (a type of satellite DNA) where a long stretch of DNA is made of repeating elements
How can we use STRs in profiling?
They can be excised using restriction enzymes and separated with gel electrophoresis
As they are often different lengths they are usually unique
What are nucleosomes?
A complex with a compacted structure created when DNA is packaged with 8 histone proteins (an octomer)
Why does DNA supercoil?
To protect DNA from damage
To allow chromosomes to be mobile during mitosis/ meiosis
What is a chromatosome?
Nucleosomes that are linked together with an extra H1 histone protein to form a string
How are less active chromatosomes structured within the DNA?
The coil into solenoid structures (~6 chromatosomes per turn)
How do long chains of solenoids fold?
In a 30nm fibre
In what form is the 30nm fibre of DNA during interphase?
Chromatin
In what form is the 30nm fibre of DNA during metaphase?
Chromosome
What is the order (from small to large) of DNA structures?
Naked DNA Nucleosome Chromatosomes Solenoid 30 nm fibre Chromatin/ chromasomes
In which structures are active genes?
Nucleosome
Chromatosomes
In which structures are inactive genes?
Solenoids
30 nm fibre
How does DNA form around nucleosomes?
Negatively charged DNA associates positively with charged amino acids on the surface of histone proteins