Unit 3 (Week 8 Nucleic Acid Structure DNA Replication and Chromosome Structure) Flashcards
What carries the genetic instructions for living things and, for example, enables a fertilized egg to develop into an embryo and eventually into an adult organism?
DNA
What provides the blueprint to create specific enzymes for the digestion of certain foods?
DNA
What are the four key criteria for something to be genetic material?
Information - The genetic material must contain the information necessary to construct an entire organism.
Replication - The genetic material must be accurately copied, a process known as replication.
Transmission - After it is replicated, the genetic material can be passed from parent to offspring. It also must be passed from cell to cell during the process of cell division.
Variation - Differences in the genetic material must account for the known variation within each species and among different species.
What is a discrete unit of genetic material composed of DNA and associated proteins?
Chromosomes. Eukaryotes have chromosomes in their cell nuclei and in plastids and mitochondria.
Greek from chromo and soma meaning colored body, refers to the early observation by microscopists that chromosomes are easily stained by colored dyes.
What is a discrete unit of genetic material composed of DNA and associated proteins?
Chromosomes. Eukaryotes have chromosomes in their cell nuclei and in plastids and mitochondria.
Greek from chromo and soma meaning colored body, refers to the early observation by microscopists that chromosomes are easily stained by colored dyes.
In the late 1920s, an English microbiologist, Frederick Griffith, studied what type of bacteria and injected mice with two strains: Type S for smooth and Type R for rough?
Pneumococci or Streptococcus pneimoniae.
What is a type of gene transfer between bacteria in which a segment of DNA from the environment is taken up by a competent cell and incorporated into the bacterial chromosome?
This was shown to happen with strep in mice.
Transformation.
How did Avery, MacLeod, and McCarty, in their experiment, prove that DNA from S type strep was responsible for transforming R type strep into S type strep?
They purified samples of DNA from S type strep, added them to mixtures of R Type bacteria and then added different enzymes such as DNase, RNase, and proteases to different mixtures to essentially denature their respective substrates. Once this was complete, and an antibody was added to remove any non-transformed R type strep, the results were centrifuged, causing all non transformed R type strep to form a pellet at the bottom.
The resulting S type was left over in each of the test tubes. The only test tube to remain “rough” was the one with DNase. This means that DNA of S type was the transforming principle in this experiment.
What is an enzyme that digests DNA?
DNase
What is an enzyme that digests RNA?
RNase
What is an enzyme that cuts proteins into smaller polypeptides?
Proteases
Explain the characteristics of the S. pneumoniae strains that made them particularly well suited for the researchers’ experiment.
Previous studies had indicated that mixing different strains could lead to transformation, or the changing of a strain into a different one. Griffith had shown that mixing heat-killed type S with living type R bacteria would result in the transformation of the type R to type S. Though mutations could cause a change of identity of certain strains, the type R to type S transformation was not due to mutation but was more likely due to the transmission of a biochemical substance between the two strains. Griffith recognized this and referred to the biochemical substance as the “transformation principle.” If Avery, MacCleod, and McCarty could determine the biochemical identity of this “transformation principle,” they could identify the genetic material for the bacteria.
What is a DNA extract?
A DNA extract contains DNA that has been purified from a sample of cells.
In the experiment of Avery, MacLeod, and McCarty, what was the purpose of using protease, RNase, and DNase if only the DNA extract caused transformation?
The researchers could not verify that the DNA extract was completely pure, that is, lacking small amounts of contaminating molecules, such as proteins and RNA. The researchers were able to treat the extract with enzymes to degrade proteins (using protease), RNA (using RNase), or DNA (using DNase). Eliminating the proteins or RNA did not alter the transformation of the type R to type S strains. Only the enzymatic degradation of DNA disrupted the transformation, indicating that DNA is the genetic material.
[Nucleic Acid Structure] What is an organic macromolecule composed of nucleotides?
Nucleic acid. The two types of nucleic acids are deoxyribonucleic acid (DNA) and ribonucleic acid (RNA).
How was the name nucleic acid coined?
When Swiss physician Friedrich Miescher in 1869 discovered a phosphorous-containing substance from the nuclei in white blood cell in waste surgical bandages, he named the substance nuclein. Later on, they were gound to be acidic molecules, which means they release hydrogen ions (H+) in solution and have a net negative charge at neutral pH. Thus, the name of nucleic acid was coined.
What is an organic molecule having three components: one or more phosphate groups, a five-carbon sugar (either deoxyribose or ribose), and a single or double ring of carbon and nitrogen atoms known as a base?
Nucleotide
What is the structure of of DNA (or RNA) formed by the covalent linkage of nucleotides in a linear manner?
Strand
What are two strands of DNA hydrogen-bonded with each other?
Double Helix. Two DNA strands are twisted together to form a structure that resembles a spiral staircase.
What allows DNA to associate into a compacted structure like a chromosome?
The wide array of different proteins to form chromosomes.
What is the complete genetic material of an organism or species?
The genome
[Review] What are the three components of a nucleotide?
A phosphate group, a 5-carbon sugar, and a nitrogen-containing base.
What pentoses are found in DNA? What about RNA?
Deoxyribose and ribose. This, their phosphate groups, and nitrogen base.
How many different bases are there and how are they categorized?
There are five different bases, only one per any given nucleotide. The bases are categorized into pyrimidines and purines.
What is either bases adenine (A) and guanine (G), which is fused double ring of carbon and nitrogen atoms?
Purines
What is any of the bases thymine (T), cytosine (U), and uracil (U), which have a single ring of carbon and nitrogen atoms?
Pyrimidines
What two bases are only found in DNA or RNA?
Thymine is only found in DNA while Uracil is only found in RNA.
Which pyrimidine(s) is (are) found in both DNA and RNA?
Cytosine
In the 5-carbon sugar molecule, what is denoted as one prime or the first carbon in the numbering scheme? Where is the 5th carbon in these pentoses?
The carbon to the right of the oxygen atom and the scheme rotates clockwise. The 5th carbon is outside of the ring of carbons.
Where are the bases and phosphate groups attached on the 5-carbon sugar molecules in a nucleotide?
Carbon 1 or one prime is attached to the base while five prime or carbon 5 is attached to a phosphate group.
What does deoxy- in deoxyribose denote in this sugar molecule?
It lacks a single oxygen atom covalently bound to the hydrogen atom off of two prime or carbon 2. In relation to ribose, which has a oxygen atom attached to the hydrogen atom off of carbon 2.
What bond in a strand of DNA or RNA links a covalent bond between the phosphate group of one nucleotide and the sugar molecule of the next nucleotide?
Phosphoester bond
Another way to look at this linkage is to notice that a phosphate group connects two sugar molecules, and from this perspective, the linkage in DNA and RNA strands is called a phosphodiester linkage, which has two phosphoester bonds.
What is the linear arrangement of phosphates and sugar molecules in a DNA or RNA strand called?
Backbone. The bases project from the backbone.
How is the backbone of a DNA or RNA strand charged?
Negatively because of the phosphate groups.
What in a DNA or RNA strand, refers to the orientation of the sugar molecules within that strand. Can be 5′ to 3′ or 3′ to 5′?
Directionality. 5’ to 3’ direction when going from top to bottom. 3’ to 5’ direction when going from bottom to top.
What is a key feature of bases based off of their structure?
The sequence.
How is directionality indicate based off of the sequence of bases?
TACG = Thymine-adenine-cytosine-guanine. For directionality, the sequence of strand is abbreviated 5’-TACG-3’
THIS is a critical feature that allows DNA and RNA to store and transmit information. These strands remain the same over time and only change in rare cases of mutation.
What was the important, experimental tool that led to the discovery of DNA double helix?
X-ray diffraction. When a substance is exposed to X-rays, the atoms in the substance cause the X-rays to be scattered.
If the substance has a repeating structure, the pattern of scattering, known as the diffraction pattern, is related to the structural arrangement of the atoms causing the scattering. The diffraction pattern is analyzed using mathematical theory to provide information regarding the three-dimensional structure of the molecule.
What other information was provided from the pattern about DNA in the x-ray diffraction technique? (2)
- Number of nucleotides per turn.
- Consistency of spacing (2-nm) between strands which corresponds to purine (A or G) bonding with a pyrimidine (T or C).
What year did James Watson and Francs Crick propose the structure of DNA and in what journal?
1953 in the journal, Nature. They discovered using ball and stick models that purine bases, A or G, bonds with pyrimidine bases, T or C.
How is the structure of DNA stabilized between base pairs on opposite strands in DNA?
Hydrogen Bonding
What is the structure in which two bases in opposite strands of DNA are held together by hydrogen bonding to each other?
Base pairs
What is the rule that refers to the phenomenon that an A in one DNA strand always hydrogen-bonds with a T in the opposite strand, and a G in one strand always hydrogen-bonds with a C?
AT/GC rule
To follow-on the AT/GC rule, what always bonds with what?
Purines (A and G) always bond with pyrimidines (T and C)
What is the distance for one complete turn of a double helix and roughly how many base pairs does it contain?
3.4 nm in length, 2 nm in width, and comprises of about 10 base pairs.
[Bonus] One nucleotide is .34 nm long hence 3.4 length for 10 base pairs.
What word is used to describe the helix structure in reference to the two strands orientating around each other?
Antiparallel
How many hydrogen bonds exist between adenosine and thymine? What about guanine and cytosine?
2 hydrogen and 3 hydrogen bonds, respectively. The bonds between guanine and cytosine are more stable since they have more bonds.
What is created on the outside of the double-helix structure based off of the asymmetrical spacing and antiparallel orientation of DNA?
A major and minor groove. These play important roles in the binding of proteins that regulate gene transcription.
What is the characteristic of the two strands of DNA that is due to the specific base pairing that occurs between nucleic acids: A pairs only with T (in DNA) or U (in RNA), and G pairs only with C.
This is a complementary characteristic.
What is the arrangement in DNA where one strand runs in the 5´ to 3´ direction and the other strand is oriented in the 3´ to 5´ direction?
Antiparallel
What is a wider groove that spirals around the DNA double helix; provides a location where a protein can bind to a particular sequence of bases and affect the expression of a gene?
Major groove
What is a groove that spirals around the DNA double helix but is smaller than the major groove?
Minor groove
What is the process by which DNA is copied? The original DNA strands are used as templates for the synthesis of new DNA strands.
DNA replication
What do you call the newly made strand in DNA replication?
Daughter strand
What do you call the original strand in DNA replication?
Parental strand
What is the correct model of DNA replication; double-stranded DNA is half conserved following replication, resulting in new double-stranded DNA containing one parental strand and one daughter strand?
Semiconservative mechanism
What is a incorrect model of DNA replication, where both parental strands of DNA remain together (are conserved) following DNA replication. The two newly made daughter strands are also joined together?
Conservative mechanism
What is another incorrect model for DNA replication, segments of parental DNA and newly made DNA are interspersed in both strands following the replication process?
Dispersive mechanism
What did Meselson and Stahl use as a model organism to determine the correct mechanism of action in DNA replication?
Bacterium, Escherichia coli. An important feature of the research involved the use of isotope labeling in Nitrogen.
Using 15N form of nitrogen (heavy) and 14N form of nitrogen (lighter), Meselson and Stahl would let bacteria grow several generations in the heavy medium first. Then, moving the bacteria to a light medium, allowed for the nitrogen source to be of light nitrogen.
Eventually they put the DNA into a gradient that would have the DNA set in a specific place in the gradient. Heaviest would sink to the bottom, heavy and light nitrogenous DNA would be around the middle, and the all light DNA would be around the top.
Using UV light, after the first round of DNA replication, all of the DNA was half-heavy meaning this data was consistent with semiconservative and dispersive mechanisms of DNA replication.
If this experiment was conducted for four rounds of DNA replication (that is, four generations), what would be the expected fractions of light DNA and half-heavy DNA according to the semiconservative model?
The expected fractions would be 1/8 half-heavy and 7/8 light.
What is the DNA strand that is used as a template for RNA synthesis or DNA replication?
Template strand. DNA replication relies on the complementarity of DNA strands according to the AT/GC rule.
What is the region called where nucleotides have access to the template strands after separation in the duplication process?
The replication fork.
After a nucleotide binds to its partner in the replication fork, what is the next bond that is formed?
A covalent bond is formed between the phosphate of one nucleotide and the sugar of the previous nucleotide.
After a nucleotide binds to its partner in the replication fork, what is the next bond that is formed?
A covalent bond is formed between the phosphate of one nucleotide and the sugar of the previous nucleotide.
T/F DNA replication produces two copies of DNA with the same sequence as the original DNA molecule.
True.
DNA replication creates two new double helixes at the same time. Where are these nucleotides attached in reference to the leading and lagging sides of the replication process?
On the leading (left) side of DNA replication, nucleotides occurs towards the direction of the fork. Whereas synthesis for the other newly lagging (right) side, occurs in small segments leading away from the replication fork.
What is the site within a chromosome that serves as a starting point for DNA replication?
Origin of replication
What happens when two DNA strands unwind at the origin of repliction and begin to unwind?
Replication proceeds outward from two replication forks.
What is a name fore this replication outwards from two replication forks at the origin of replication? In other words, The process in which DNA replication proceeds outward from the origin in opposite directions.
Bidirectional replication.
T/F Eukaryotes and bacteria have the same amount of origins of replication.
False. Bacteria DNA genome is rather small so it has one but Eukaryotes have many so replication can happen in a reasonable amount of time.
What completes the replication process in eukaryotes?
When the newly made strands from each origin make contact with each other.
What is a template understanding of what needs to happen in DNA replication? (2)
The strands of the double helix must separate.
The resulting fork must move.
What enzyme uses ATP to separate DNA strands during DNA replication?
DNA helicase
In what direction does DNA helicase move when it is bonded to one of the DNA strands which allows and how does it use ATP?
It travels in the 5’ to ‘3 direction and uses ATP to break hydrogen bonds between base pairs.
The enzyme, DNA helicase, can create knots ahead of the replication fork called supercoils. How are these removed?
By another enzyme called DNA topoisomerase.
How does DNA topoisomerase travel in relation to the replication fork?
Slight ahead of the fork looking for knots.
While the two template DNA strands have been separated, they must remain so until their daughter strands have been made – What is the proteins that binds to both of the single strands of parental DNA and prevents them from re-forming a double helix during DNA replication?
Single-strand binding protein
What is the job of DNA gyrase?
In bacterial cells, it is an essential enzyme that catalyzes the ATP-dependent negative super-coiling of double-stranded closed-circular DNA.
Positive super force coiling is in right handed direction which is the same direction as the double helix in DNA.
Gyrase protein binds to a supercoil molecule and creates a double break through ATP hydrolysis. The break in the strand is repaired and gyrase releases leaving a negative supercoil in bacterial DNA.
When single-strand binding proteins bind to each separated strand of DNA replication, what does it accomplish?
It allows the bases within the template strands to be exposed so they can act as templates for the synthesis of complementary or daughter strands.
What enzyme is responsible for covalently linking nucleotides together during DNA replication?
DNA polymerase
It was discovered in the 1950s and resembles a human hand with the DNA threaded through it.
What strand can DNA polymerase only add nucleotides to when helicase separates the strands?
Only the free 3’ (three prime) end.
In order for DNA polymerase to begin building the daughter strand, a short primer known as RNA primer is synthesized first giving DNA polymerase a starting point.
What are the individual nucleotides with three phosphate groups called which utilize the catalytic site on DNA polymerase to bind to exposed bases via hydrogen bonds in synthesis of a daughter strand?
Deoxynucleoside triphosphates
Referring to the catalytic site on DNA polymerase and the interaction with deoxynucleoside triphosphates, what is accomplished by the active site on the enzyme? (2)
- Bond is broken between the first and second phosphate.
- Attaches resulting nucleotide with one phosphate group (a deoxynucleoside monophosphate) to the 3’ end of a growing strand via a phosphoester bond.
What is significant in the breaking and bonding of the deoxynucleoside via the work of DNA polymerase in regards to net free energy?
This process is exergonic because of the covalent bond breakage which created pyrophosphate. The energy released is used for adjacent nucleotides.
Pyrophosphate is then broken down into two phosphates.
Does the oxygen in a new phosphoester bond come from the sugar or from the phosphate?
The oxygen in a new phosphoester bond comes from the sugar.
[Bonus] What are the rates of DNA polymerase synthesis of DNA in bacteria? What about eukaryotic cells?
500 nucleotides per second in bacteria; 50 nucleotides per second.
What are the other two enzymatic features of DNA polymerase?
- It can elongate a preexisting strand by making DNA.
2. it can synthesize new DNA only in a 5′ to 3′ direction. Antiparallel to the template or parent strand.
Since DNA polymerase cannot begin DNA synthesis on a bare template strand, what is needed to begin its own DNA replication process?
DNA primase. This enzyme is the one that synthesizes a primer for DNA replication.
It is a short segment of RNA, typically 10 to 12 nucleotides in length. This “primes” the DNA replication process.
During DNA replication, what DNA strand made in the same direction that the replication fork is moving? The strand is synthesized as one long continuous molecule.
Leading strand
During DNA replication, what DNA strand made as a series of small Okazaki fragments that are subsequently connected to each other to form a continuous strand?
Okazaki fragments
Short segments of DNA synthesized in the lagging strand during DNA replication.
Lagging strand
What must the leading strand do in order for the lagging strand to add another Okazaki fragment?
At what point is the first and second Okazaki fragments connected?
Elongate which is a property of DNA polymerase.
After the third fragment is added.
Which strand, the leading or lagging strand, is made discontinuously in the direction opposite to the movement of the replication fork?
The lagging strand is made discontinuously in the direction opposite to the movement of the replication fork.