Unit 3 Flashcards
Energy and Metabolism (ch 8,9,10)
Can you describe the structure of a nucleotide?
1 phosphate group
1 sugar (a pentose)
1nitrogenous base
“nucleoside” = nitrogenous base + sugar
How do ribonucleotides and deoxyribonucleotides differ?
deoxy = missing an O
= DNA a 2’ carbon attached to 2 H
RNA has a 2’ carbon attached to a OH and a H groups
Can you describe the structure of a polynucleotide?Mention phosphodiester bond and sugar-
phosphate backbone.
chain of many phosphate-sugar-phosphate-sugar
dehydration synthesis rxn occurs:
phosphate group + sugar of another nucleotide
= phosphodiester bond
2 phosphate groups must be released to provide energy to make the phosphodiester bond
Can you describe how a polynucleotide has a 5’ to 3’ orientation?
one side ending with a phosphate group
= 5’ end
one side ending with the -OH group on the sugar groups
= 3’ end
Can you describe complementary base pairing in nucleic acids? Mention the type of bond involved
and how many bonds are between each base pair.
RNA can associate with itself in complementary base pairing
the 2 strands of DNA can associate by complementary base pairing
-> Hydrogen bonds between the nitrogenous bases
-> Cytosine + Guanine = 3 H-B
-> Adenine + Thymine = 2 H-B
Complementary base pairing in DNA occurs between what types of nitrogenous bases? (structure)
between a pyrimidine and a purine
Can you describe the levels of structure in RNA (up to quaternary structure)?
Can you list the characteristics of DNA that allows it to act as genetic material? Describe the types
of information that is stored in DNA.
Can you discuss the role of complementary base pairing in the functions of DNA. Which bases are
purines? Pyrimidines?
pyrimidine = 1 ring (cytosine, thymine, uracil)
purines = 2 rings (adenine, guanine)
Can you describe some of the many functions of RNA? Compare coding vs. non-coding RNAs. Explain
why RNA is capable of producing many functional classes (similar to proteins)
What is Chargaff’s Rule?
% of A is equal to the % of T
% of G is equal to the % of C
How did Watson, Crick and Franklin’s discovery of the double-stranded nature of DNA explain Chargaff’s Rule ?
DNA has a helical shape
the double helix has an uniform diameter
= shows that purines must be paired with pyrimidines
= shows that one strand can be used as template
Can you describe the components of Watson and Crick’s model of DNA’s secondary structure?
they discovered:
- the helical shape of DNA
- the width of the helix
- the spacing between nitrogenous bases
Explain the Hershey and Chase experiment
was the hereditary material DNA or protein?
- bacteriophage injects one batch with radioactive sulfur and a second batch with radioactive phosphate
- each batch is mixed with bacteria cells in a blender
- the phage DNA enters the bacteria cell
- the mixture is centrifuged
- the bacterial cells is heavier = sink in the bottom
- the bacteriophage (“virus”) floats
results
1st batch: radioactive substance (pink) is floating
2nd batch: radioactive substance (blue)is sinking
= blue entered the bacteria cells and pink stayed outside
= DNA entered the bacteria cells and proteins stayed outside
= DNA is hereditary and not proteins
Why were radioactive sulfur and radioactive phosphate good tags?
good tags because phosphate is only found in dna and sulfur is only found in protein
Explain the manipulations in the Meselson-Stahl experiment
- bacteria from a medium with N15 were cultured
- dna was synthesized from N15
- bacteria was transferred to a medium with N14
- dna was centrifuged after 1 dna replication
- bacteria sample was collected
- dna was centrifuged after a 2nd dna replication
- bacteria sample was collected
Explain the observations in the Meselson-Stahl experiment
after 1 replication
half of the dna molecule is heavy and half is light
after 2nd replication
half is light and half is heavy or it is all light
Explain the conclusion in the Meselson-Stahl experiment (which theory of dna replication)
3 theories of dna replication
1. conservative = dna gets copied and make a new dna
2. dispersive = dna gets cut at various part and each cut would get copied and then reattach to make a new dna
3. semi-conservative = dna strands separate and each serve as a template to copy a second strand and producing 2 dna molecules
conclusion
= semi conservative dna replication
To form the phosphodiester bonds in DNA polymerization, where does the energy come
from?
the energy required for the formation of phosphodiester bonds is provided by the dephospho rylation of nucleoside triphosphate.
= removing 2 phosphate groups from the nucleoside
Can you describe the function of a DNA polymerase? Include the specific rules that it follows: can it
initiate polymerization? In what direction does it synthesize a new DNA strand?
DNA polymerase cannot initiate synthesis of a polynucleotide strand
it can only add nucleotide to the 3’ end of an existing polynucleotide strand
so primase comes in 1st, adds the short chain of RNA, then DNA polymerase comes in
Can you define a replication origin?
specific sequence of nucleotides that tell te enzymes to start replication
Can you define a replication bubble?
proteins recognize the sequence of the replication origin and attach to the DNA.
once attached, it separates the two strands and creates a replication bubble
Can you define a replication forks?
there is a replication fork at each end of the replication bubble
= a Y shaped region where replication occurs
Can you define a bidirectional replication?
replication occurs in both direction from the origin
Can you sketch a replication fork?
o Label the leading strand template and indicate its orientation (using 5’ and 3’ as markers).
o Label the lagging strand template and indicate its orientation.
o Use arrows (one for continuous synthesis and multiple for discontinuous synthesis) to
indicate the direction of leading and lagging strand synthesis.
o Indicate the orientation of the newly synthesized DNA.
Can you describe the machinery (enzymes and proteins) and process for each of the following:
1. Opening the double helix at the replication fork. (Helicase, SSBP, Topisomerase, Primase +
RNA nucleotides)
2. 3. Synthesis of the leading strand (DNA Pol III + DNA nucleotides)
Synthesis of the lagging strand (synthesis of Okazaki fragments, Primase, DNA Pol III, RNase
H, DNA Pol I, DNA ligase)
- unzip the double-stranded DNA & keeping it open
-> helicase
=unzip
-> SSBP
= keep them from reannealing = zipping back
Describe the fourth step of DNA polymerization
- synthesis of the new complementary DNA strand
-> DNA polymerase III
= adds complimentary DNA nucleotides to the free 3’ end of the strand
-> Sliding clamp
= makes sure DNA pol III stays in contact with the template strand during polymerization. it is right next to the DNA pol III.
DNA pol II can only add nucleotides to an open 3’ end
= needs the help of the leading strand and lagging strand
What is a lagging strand?
DNA pol III synthesizes the leading strand, then works along the other template strand
= forms a new DNA strand called lagging strand
-> replicates itself 1 section (“Okazaki fragments”) at the time
-> in the opposite direction from the movement of the replication fork
What is a leading strand?
DNA pol III synthesizes a complementary strand in the 5’ -> 3’ direction
= in the same direction of the replication fork
this makes a new DNA strand called the “leading strand”
What is a primer ?
the primase forms a primer, a short chain of NA that is added to the parental DNA strand.
it uses the parental DNA as a template
Complementary DNA strands can only be
elongated in the ___ → ___ direction.
Complementary DNA strands can only be
elongated in the 5’ → 3’ direction.
how many RNA primer(s) are required for each leading strand? for each lagging strand?
leading strand = one primer
lagging strand = one primer for each Okazaki fragments
Describe the second step of DNA polymerization
- release the tension
-> topoisomerase
=relieve strain by uncoiling the strand on the left of the replication fork
Describe the third step of DNA polymerization
- start synthesizing the new strand
-> primase
= forms a “primer”
-> DNA polymerase
= catalyzes the polymerization of DNA nucleotides
Describe the fifth step of DNA polymerization
- remove & replace RNA primers w/ DNA nucleotides
-> Rnase H
= recognizes RNA-DNA hybrid segments
= degrades the RNA primer by hydrolysis of the phosphodiester bonds
-> DNA polymerase I
= adds DNA nucleotides to free 3’ ends of leading/lagging strands
Describe the sixth step of DNA polymerization
- glue fragments of lagging strands together
-> DNA ligase
= attach fragments of DNA on the new strands
= join 3’ end of one to the 5’ of another fragment
= forms a continuous polynucleotide strand
Can you define the replisome (replication factory) and briefly discuss how this factory increases the
efficiency of DNA replication?
Can you compare other differences between prokaryotic and eukaryotic replication?
prokaryotes
= 1 origin of replication
= circular genome
= uses only DNA pol III and I
= longer Okazaki fragments
= no telomeres
eukaryotes
= linear genome
= more than 1 origin of replication
= much more base pairs in its DNA = takes longer
= uses over 11 different DNA pol
= shorter Okazaki fragments
= has telomeres
Why would eukaryotic chromosomes need telomeres? What are telomeres? What do they prevent
in linear chromosomes?
telomeres
= short repeated DNA sequence at both ends of a linear DNA strand
= it contains no genes
= get shorter with each replication event
-> so eukaryotes can’t live forever, bc the telomeres would disappear at some point
What do telomeres prevent in linear chromosomes?
- prevent activation of DNA damage signalling
- protects against the organism’s genes shortening
What is telomerase? How is this enzyme implicated in the difference between replication in somatic
cells versus germ line cells?
an enzyme that extends the length of telomeres
it is found in
germ cells
cells in embryos/fetuses
tumor cells
it is active all the time in germ cells, whereas other cells don’t have telomerase.
Telomeres acts as a protection against cancer, how?
shortening of telomeres protect against cancer because it limits the number of division that somatic cells can undergo
bc cancer cells = unlimited cell division
Can you list the reasons why polymerization of DNA by DNA polymerase limits mistakes? Why this is
important?
Can you describe the mechanism of proofreading by DNA polymerase?
DNA pol:
- proofreads each nucleotide against its template
- replaces wrong nucleotide by the right one
this is possible bc DNA pol has exonuclease abilities
= it can cleave off nucleotides
Can you describe some of the many types of damage that DNA sustains?
reactive/harmful chemicals
radioactivity
x-rays
uv light
Describe the damage of UV light to DNA
= it links two adjacent pyrimidines and forms thymine dimer)
= it distorts the DNA molecule
= it can cause the disease xeroderma pigmentosum
what is xeroderma pigmentosum
-> inherited disease that causes hypersensitivity to sunlight
->mutations caused by UV light
-> mutations lead to defect in the excision repair mechanism, which is supposed to repair the damage of UV light
= damages of UV light are left unresolved
-> defect in the excision repair mechanism
-> mutation of the skin
Can you describe how DNA damage such as a mismatch base pair created by mistake during DNA
replication or an external agent can lead to a base pair substitution mutation after cell division?
Can you describe DNA excision repair? Mention general steps, and types of enzymes required in
each step.
- mismatched base pairs are detected
- nuclease
= makes 2 cuts on the damaged strand & releases the damaged section - DNA polymerase
= places the correct nucleotides - DNA ligase
= seals everything together
What is DNA methylation? What are some of its functions?
put a tag on the parental strand by adding a methyl group
= it allows for fast repair
bc mistakes in DNA tend to happen when you are making a new parental strand has already gone trough spell check etc so it should be mistake free
functions:
1. so we can know which strand has the right sequence and which one is mistaken:
- > parental DNA is methylated & daughter DNA isn’t
2. methyl group helps protecting the DNA by getting in the way of the binding site of the enzyme that wants to cut the DNA
What causes DNA dimerization? What is an example of dimerization?
UV light exposure
thymine dimer
What is it important for DNA in prokaryotes and eukaryotes to be properly packaged?
to prevent bad enzymes from digesting the DNA and prevent the DNA from breaking