Lecture 7 - The structure of DNA Flashcards

1
Q

Who was Phoebus Levene and what theory did he propose? (1909)

A

Tetranucleotide theory

Showed each building block of DNA is a nucleotide:

Phosphate group linked to a deoxyribose sugar, linked to nitrogenous base

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2
Q

What sugar is used in DNA and RNA?

A

DNA: pentose (5C) deoxyribose
RNA: pentose (5C) ribose

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3
Q

What is the difference in structure between ribose and deoxyribose molecular structure?

A

Hydroxyl group in ribose on 2’

Hydrogen in deoxyribose on 2’

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4
Q

What bases are purines?

A

Adenine

Guanine

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5
Q

What bases are pyrimidine?

A

Cytosine

Thymine

Uracil

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6
Q

What are nucleosides and where/what is the bond?

A

Sugar and base

Glyosidic bond: between C-1’ and N-9 (purine) or N-1 (pyrimidine)

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7
Q

What are nucleotides and where/what is the bond?

A

Phosphorylated nucleosides

Ester links: between sugar C-5’ group and the phosphate

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8
Q

What forms do nucleotides come in?

A

All come in mono, di and tri phosphate form

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9
Q

What kind of molecule is DNA?

A

Polynucleotide with polarity

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10
Q

What is the polarity associated with DNA?

A

Phosphodiester bond links the 3’C of one nucleotide to 5’ of the next

Base sequence is written and read 5’ to 3’

Polarity at the 5’ phosphate end –> 3’ hydroxyl end

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11
Q

How did Levene ‘tetranucletoide model’ not support Avery et al?

A

Showed how DNA was simple and repetitive and could not be the genetic material

Not believed to be transforming principle

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12
Q

Who was Erwin Chargaff and what did he propose?

A

Nucleotides in DNA should be present in equal proportions

(If Levene was correct)

%T = %A = %G = %C

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13
Q

How did Chargaff show Levene to be incorrect?

A

Measured concentrations of each of 4 nitrogenous bases in different organisms

Different organisms had different DNA constitutions

Reality: %T = %A and %G = %C

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14
Q

What did Linus Pauling say about DNA? (1951)

A

Described the alpha helix

Basic structure present in many proteins

Used X-ray crystallography - so shape must be helical

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15
Q

How does an X-ray crystallography indicate the helical structure of DNA?

A

Crystalline target molecule diffracts X-rays

Causes exposed patches on photographic film

Resulting diffraction pattern is unique

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16
Q

What did Huygens show with the ‘Wave theory of light’?

A

When light passes through a small opening, a wave front is propagated on the other side, single spot appears on a screen

Wider slit, all points across the slit act as point source - results in single slit diffraction pattern on screen

2 slits causes interference - causes double slit interference pattern on screen

With a diffraction grating the pattern becomes much sharper

17
Q

What did Augustin Fresnel extend the ‘Wave theory of Light’ concept?

A

Showed diffraction occurred around a solid object with the same width as a slit

Replacing slit with object of same size causes the same results

18
Q

What are the spatial relationships with the light theory?

A

Features that are close produce widely separated reflections

Features that are distant produce closely separated reflections

Vertical grids = horizontal spots
Grid = cross

19
Q

What did Maurice Wilkins do?

A

Stretched DNA and air dried it

Allowing for stretching into long fibres and mounting in front of X-ray source

20
Q

What was Maurice Wilkins produce?

A

1950 - Produced X-ray diffraction of dried DNA

High resolution

Without clear model in mind - too difficult to interpret

21
Q

What was Gosling and Franklin B model of DNA?

A

1952 - Used hydrated DNA after 100 h of exposure

Cross is unmistakable - DNA must be helical

22
Q

Who proposed the triple helical model of DNA?

A

Linus Pauling

Triple helix 3-strand model, with bases pointing outwards

(supported by Watson and Crick)

23
Q

What did Watson, Crick and Wilkins propose?

A

Double helix 2-strand model, with the bases pointing inwards

24
Q

What was each turn of the DNA helix measured to be?

A

3.4nm

(Spots far apart give features that are close together)

25
How many bases were there thought to be per turn of the DNA helix?
10 bases per turn (More distant the spots, smaller the actual distance in the target)
26
What was the measured diameter of DNA to be?
2nm Calculated using degrees of rise within the 'X'
27
Why did Pauling model of DNA fail?
(triple helix) The negative charges of the stacked phosphate groups would repel each other and destabilise the molecule - Sugar phosphate backbone must be compressed together
28
How did Watson's deduction support Chargaff rule?
2 purines = too wide for DNA 2 pyrimidines = too narrow for DNA = A must pair with T = G must pair with C
29
What was Watson and Crick's model in 1953?
Made DNA with metal scraps, almost 2m tall Watson: specific A/T and G/C pairing scheme Crick: idea of antiparallel strands Everything clicked into place beautifully
30
What are 6 key features of Watson-Crick model?
1. Right-handed (clockwise) double helix 2. The strands are anti-parallel 5' --> 3' and 3'--> 5' 3. Sugar-phosphate backbones are on the outside of the helix, bases oriented towards the central axis 4. Complementary base-pairing - bound by weak hydrogen bonds 5. Base pair distance (10.5 bp per turn, helix turn = 3.6nm 6. major and minor grooves - backbone not equalling spaced causing grooves
31
How many bonds does A-T have?
2 hydrogen bonds
32
What are the other structural variants of DNA?
A-DNA: occurs in low hydration conditions (uncertain whether this occurs) B-DNA: most structurally stable Z-DNA: taken up physiologically by stretches of alternating pyrimidines and purines