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Genetics > DNA Replication > Flashcards

DNA Replication Flashcards

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1
Q

What were people uncertain genetic material was in the early 20th century?

A

Carbohydrates
protein
lipid
something else

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2
Q

Recognition that _ was genetic material was a key insight

A

DNA

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3
Q

What did DNAs structure immediately suggest?

A

DNA could serve as a blueprint and that blue print could be passed down through generations

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4
Q

Where are genes carried?

A

chromosomes

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5
Q

What are chromosomes comprised of?

A

DNA and proteins

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6
Q

Experiements suggested that the _ component of chromosomes is the genetic material

A

DNA

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7
Q

What was Fredrick Griffith’s experiement with?

A

Mice and streptococcus pneumoniae

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8
Q

What cells killed inoculated mice?

A

Smooth cells
S-strain

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9
Q

What cells did not kill inoculated mice

A

Rough cells
R-strain

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10
Q

How do you eliminate the virulence in the s-strain

A

with heat

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11
Q

What happens to a mouse with live R-strain cells and heat-killed S-strain? and what was recovered?

A

Mouse dies
Live S-strain was recovered

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12
Q

What does recovering live S-strains from dead mice tell us?

A

Cell debris from dead S-cells had converted R cells into S-cells

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13
Q

Avery-Mcarty-Mcleod

What material in the cell debris was responsible for the transformation of the S cell DNA into the live R cells?
How do we know?

A

DNA
The mouse did not live until the DNA debris was destroyed

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14
Q

What was the first evidence that DNA was genetic material?
(From Griffith, Avery-mccarty-mcleod experiment)

A

The mouse with live R-cells and dead S-cells did not live until the S-cell DNA debris was destroyed

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15
Q

What was the Hershey-Chase experiement?

A

Phages with radiolabeled protein and phages with radiolabeled DNA were used to infect hosts. The host cells were then processed to see if the radiolabel was within cells or outside of cells. Radioactive DNA was recovered in bacteria due to the radiolabeled DNA.

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16
Q

What was recovered in the Hershey-Chase experiment?

A

Bacteria with radioactive DNA

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17
Q

Structural features of DNA must allow for _ _

A

Faithful replication

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18
Q

Genetic material must have _ _

A

informational content

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19
Q

Genetic material _ be able to change on rare occaision

A

must

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20
Q

What 3 pieces of information about DNA
were crucial in determining its structure

A

The chemical composition of DNA
Chargaff’s rule
X-ray crystallography

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21
Q

What are the 3 components of DNA?

A

Tri-hosphate
a deoxyribose sugar
A nitrogenous base

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22
Q

What are the 4 nitrogenous bases of DNA

A

Purines: adenine and guanine
Pyrimidines: cytosine and thymine

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23
Q

Purine structure?

A

double ring

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24
Q

Pyrimidine structure?

A

single ring

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25
Q

What is chargaffs rule?

A

T=A, C=G but A+T does not necesarily = C+G

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26
Q

Who conducted x-ray crystallography of DNA

A

Rosalind Franklin and Maurice Wilkins

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27
Q

What did x-ray crystallography suggest?

A

DNA is long, skinny, helical and comprised of two similar parts

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28
Q

What happened in approx 1953?

A

James watson and francis crick inferred that DNA is a double helix

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29
Q

Watson and Crick
“_ must always pair with _ because…”

A

Purine must always pair with a pyrimidine because it is most consistent with xray data

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30
Q

What are DNA’s complementary base pairs?

A

A-T
C-G

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31
Q

What holds the strands of DNA together

A

Hydrogen bonding between the nucleotides

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32
Q

How many Hydrogen bonds does A-T have?
C-G?

A

AT-2
CG-3

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33
Q

Where do base pairs stack in DNA?

A

In the center of DNA as planar structures

34
Q

Do major and minor grooves impact its function?

A

yes
Alternating minor and major

35
Q

Meselson-Stahl experiment found that DNA replication is _

A

Semi-conservative

36
Q

What does semi-conservative replication mean?

A

1 old strand and 1 new strand

37
Q

When do you see the replication fork?

A

Second round of replication

38
Q

Replicating DNA is mixed between _ and _ states

A

unwound and overwound states

39
Q

_ _ removes twists and supercoils to make replication possible

A

DNA gyrase

40
Q

_ unwinds double helix at the replication fork and SSB binds

A

Helicase

41
Q

_ _ cuts DNA strands and rotates them to prevent supercoiling

A

DNA gyrase

42
Q

What rejoins the strands

A

DNA gyrase

43
Q

_ help relive tension in DNA

A

Topoisomerases
(gyrase)

44
Q

What do topoisomerases do?

A

Break the phosphodiester bonds between nucleotides and then reseal these breaks

45
Q

E.coli

Initiation of replication begins at the ___

A

Origin (oriC)

46
Q

E.coli

DnaA binds to _ _ which are specific _ _ _

A

DnaA binds to DnaA boxes, which are
specific, short DNA sequences

47
Q

E.coli

What is adjacent to DnaA boxes?

A

AT-rich sequences that melt easily which opens up the DNA

48
Q

E.coli

What do additional DnaA molecules do once DNA is opened up a bit?

A

additional DnaA molcules bind to the DNA less specifically

49
Q

E.coli

Opening up of the DNA leads to the recruitment and loading of _

A

Leads to the recruitment and loading of DnaB (helicase)

50
Q

E.coli

Once bound _ (DnaB) further _ _ _

A

Once bound helicase (DnaB) further opens the DNA

51
Q

E.coli

After helicase further opens up DNA this leads to the recruitment of the _

A

replisome

52
Q

E.coli

What does the replisome do?

A

It allows for DNA polymerase to come in and synthesize a complementary DNA strand

53
Q

E.coli

What is DNA polymerase?

A

The enzyme that synthesizes a complementary strand of DNA

54
Q

E.coli

Who isolated DNA polymerase from E.coli and demonstrated its function in vitro

A

Arthur Kornberg

55
Q

E.coli

How many DNA polymerases does E.coli have?

A

5

56
Q

E.coli

DNA pol I catalyzes chain growth _ - _ end

A

5’ to 3’

57
Q

E.coli

DNA pol I additional 3’ to 5’ exonuclease activity

A

Removes mismatched bases

58
Q

E.coli

DNA pol I additional 5’ to 3’ exonuclease activity

A

Degrades single strands of DNA or RNA

59
Q

E.coli

Which DNA pol does most of the work in replication in E.coli ?

A

DNA pol III

60
Q

E.coli

What are the two types of strands at the replication fork?

A

Leading and Lagging

61
Q

E.coli

Which is the leading strand?
Which is the lagging strand?

A

Leading strand (3’) is going into the fork
Lagging strand (5’) is going away from the fork

62
Q

DNA synthesis is primed using _ copied from DNA generated by _

A

Using RNA copied from DNA generated by primase. This gives DNA pol III a place to start

63
Q

Which DNA pol extends the with new DNA?

A

DNA pol III

64
Q

What does DNA pol 1 do to the RNA at the 5’ end?

A

Remove the RNA and fills the gap

65
Q

What connects the adjacent fragments after the RNA (primers) are removed?

A

DNA ligase

66
Q

Normal base pairing results in…

A

faithful transmission of the DNA

67
Q

Mistmatches between bases can occur, sometimes resulting in

A

mutations

68
Q

Point mutations

What is a transition?

A

When one purine changes to the other
or when one pyrimiding changes to the other

69
Q

Point mutations

What is a transversion?

A

When a pyrimidine transitions to a purine or vice versa

70
Q

What activity enables error correction?

A

3’ to 5’ exonuclease activity

71
Q

What facilitates DNA replication on both strands?

A

Replisome, accessory proteins and looping

72
Q

Eukaryotes

How do eukaryotic origins differ from e. coli origins?

A

Eukaryotes have multiple replication origins. Different parts of the chromosome will be replicated simultaneously

73
Q

Eukaryotes

How does replication extend from these origins?

A

Bidirectionally

74
Q

Eukaryotes

Eventually contigious, fully replicated _ are produced

A

Chromatids

75
Q

Eukaryotes

When does DNA replication occur in eukaryotes?

A

During S phase

76
Q

Eukaryotes

Initiation of DNA replication at the eukaryotic involves _ of specific .
Such as _

A

Involves recruitment of specific proteins.
Such as Cdc6

77
Q

Eukaryotes

What happens after the specific proteins are recruited?

A

Helicase will unwind the DNA, allowing DNA polymerase to bind

78
Q

Eukaryotes

What melts the AT regions to help binding to chromosome?

A

Helicase + Cdt1

79
Q

Eukaryotes

How do you fill the gap at the teleomere in linear chromosome?

A

Telomerase helps extend the telomeres, fixing the gap
(most of the gap)

80
Q

Eukaryotes

What happens after the telomere is extended by telomerase?
Example in humans?

A

Proteins bind to the end to form a cap that stabilizes the structure at the end.
TRF 1 and TRF 2

Genetics (17 decks)

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