DNA Replication, Transcription, and Translation Flashcards
What is the central dogma of molecular biology?
DNA makes RNA, RNA makes protein
What are the two kinds of nucleic acid?
DNA and RNA
How do DNA and RNA differ?
in chemical composition and structure
- RNA is single-stranded; DNA double
- RNA is made of ribose; DNA of deoxyribose (missing an OH group that RNA has)
- RNA contains uracil; DNA contains thymine
Who first suggested a double-helical model for the structure of DNA?
James Watson and Francis Crick
How did they figure out that DNA must have a double-helix structure?
best way molecule could fit together from the evidence they had (including x-ray crytallography)
What was the chemical evidence about the structure of DNA?
puring and pyrimiding nigrogenous bases; deoxyribose sugar, phosphate
What is the biological evidence about the structure of DNA?
ratios between bases (proportion of A = T; C = G)
What is the physical evidence about the structure of DNA?
x-ray crystallography; 2 nm diameter, double helical structure
What does the fact that nitrogenous bases are located on the inside of a DNA molecule indicate about their nature?
they’re relatively hydrophobic
What deductions did evidence from Rosalind Frankland’s work enable Watsom to make?
- DNA was helical
- width of the helix
- spacing of the nitrogenous bases
- nitrogenous bases on inside
What is Chargaff’s rule?
the proportion of A = T; C = G
Does A + C have to equal G + C?
no
What are the two types of nitrogenous basees?
pyrimidines and purines
What is the difference between a pyrimidine and a puring?
purines are made of two carbon-nitrogen ring bases (4 N in total); pyrimidines are made of one nitrogen ring base (2 N in total)
What type of nitrogenous base is adenine?
purine
What type of nitrogenous base is thymine?
pyrimidine
What type of nitrogenous base is guanine?
purine
What type of nitrogenous base is cytosine?
pyrimidine
What two components of a DNA molecule interact to form the phosphate-sugar backbone of DNA?
the free 3’-OH group of deoxyribose in the 1st nucleotide and the 1st 5’ phosphate in the 2nd nucleotide (5’P - 3’OH)
How many nucleotides may be present in a single DNA molecule?
many millions
How many strands is a DNA molecule made of?
2
How are the two strands of a DNA molecule oriented with respect to each other?
antiparallel (one strand goes from 3’ to 5’; other from 5’ to 3’)
What are the base pairings in a DNA molecule?
A:T and C:G
How many hydrogen bonds are present between adenine and thymine?
2
How many hydrogen bonds are present between cytosine and guanine?
3
In what direction does a DNA molecule grow?
from 5’ to 3’ (must add 5’P to free 3’ OH)
What do restriction enzymes do?
break DNA phosphodiester bond at a recognized “binding site”
How did evidence indicate that a purine and a pyrimidine had to pair (instead of purine-purine or pyrimiding-pyrimidine)?
process of elimination - knew that the pairing had to be 2 nm wide; only pair that fit that was purine-pyrimidine?
How did evidence indicate that A paired with T and C paired with G?
only pairings that allowed bases to hydrogen bind
What would the diameter of a DNA molecule be if a pyrimidine bonded to another pyrimidine, and how does that relate to the X-ray data?
less than 2.0 nm (too thin compared to X-ray data)
What would be the diameter of a DNA molecule if purines bonded to pyrimidines, and how does that relate to the X-ray data?
larger than 2.0 nm (too thick compared to X-ray data)
How do the amount of DNA per cell and the number of sets of chromosomes per cell relate?
by a precise correlation
How does the molecular composition of DNA compare between all the different cells of an organism?
it’s the same
How does the composition of both RNA and proteins compare between different cell types?
highly variable
How does the stability of DNA and RNA/proteins compare?
DNA is more stable, while RNA and proteins are synthesized and degraded quite rapidly in living organisms
Why is DNA more stable than RNA?
it’s double-stranded and lacks an OH group that could be acted upon
What were the three possible models for DNA replication?
conservative, semiconservative, and dispersive
What does the conservative model of DNA replication predict the products of the first and second replication will be?
first replication: one DNA molecule of both template strands; one of both copied strands
second replication: 3 molecules of all copied strands; 1 molecule of original template strands
What does the semiconservative molel predict the products of the 1st and 2nd replication will be?
1st: 2 molecules each of 1 template and 1 copied strand
2nd: 2 molecules with 1 parent and 1 copied strand; 2 with just copied strands
What does the dispersive model predict the products of the 1st and 2nd replication to be?
1st: 2 molecules with bits of template in each strand
2nd: 4 of the same
What experimental procedure was used to show that the semiconservative model was correct?
bacteria were cultured in a medium with a heavy N isotope, then transferred to a medium with a lighter isotope; the DNA sample was then centrifuged after the 1st and 2nd replications
How did centrifuging the products from the 1st and 2nd replications from the bacteria DNA replication experiment support the semiconservative model?
more dense DNA settled to the bottom while less dense settled on top; the results only matched the semiconservative model
What is the origin of repliaction?
where DNA replication starts on a chromosome
How many origins of replication are found on a circular choromosome?
1
How many origins of replication are found in a eukaryotic chromosome
many
Why can 1 strand of DNA serve as a remplate for a new one?
the 2 strands are complimentary
Why do circular chromosomes not have the same problems with replication as eukaryotes?
eukaryotic chromosomes are linear, so it’s not possible to replicate all strands in 5’ to 3’ direction (whereas in circular, can just continue around molecule in correct direction until it’s all done)
also, linear chromosomes end up losing some of their end each time (primers added)
How is the leading strand synthesized?
continuously, moving toward the replication fork
How is the lagging strand synthesized?
as a series of segments (okazaki fragments)
Why is a primer required for repliacion?
DNA polymerases can’t initiate synthesis (on a single strand) - need something to build on
What is a primer?
short stretch of RNA, synthesized by primase, using parental DNA strand as template
In what direction can a new strand elongate in replication and why?
5’ to 3’ since polymerase can only add nucleotides to free 3’ end of growing strand
What does helicase do?
enzyme that untwists double helix at replication fork
What do single-strand binding proteins do?
bind/stabilize by replication fork during replication - prevent retwisting
What is another name for topoisomerase?
gyrase
What does topoisomerase do in replication?
corects “overwinding” ahead of forks by breaking, swiveling, and rejoining DNA strands
What does primase do in replacation?
makes the primer
How many RNA primers are needed to synthesize the leading tsrand?
1
What happens after the lagging strand is synthesized in fragements?
DNA polymerase I replaces RNA primer with DNA; DNA ligase joins sugar phosphate backbones of fragments into continuous strand
How long is each okazaki fragment?
1000-2000 nucleotides long
How many RNA primers does the lagging strand need?
one for every Okazaki fragment
What model does DNA polymerase follow to synthesize DNA/
replication machine - “reels” in parental DNA and “extrudes” newly-made daughter DNA molecules (assisted by other proteins)
What occurs immediately after DNA synthesis?
proofreading
WHat enzyme proofreads DNA?
DNA polymerase
What can happen in DNA polymerase misses mistakes?
- mismatch repair (other enzymes correct)
What kind of mistakes can be corrected in DNA proofreading?
DNA damage by expoure to harmful chemical or physical agents or other spontaenous changes
What is nucleotide excision repair?
nuclease cuts out and replaces damaged stretch of DNA
What does DNA ligase do?
rebuilds the sugar-phosphate backbone of DNA whenever DNA is cut and nucleotides are replaced
What is the error rate after prroofreading?
Low but not zero
What happens when DNA sequence changes become permanent?
can be passed onto next generation (mutations)
What are the source of genetic variation upon which natural selection operates?
mutations (changes in DNA sequence)