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Transcript
4/3/2013
Nucleotides and DNA Structure
C483 Spring 2013
1. Purine(s) which are found mainly in both deoxyribonucleotides and ribonucleotides are A) thymine and cytosine.
B) cytosine and uracil.
C) cytosine.
D) guanine and cytosine.
E) adenine and guanine.
2. The abbreviation dGp indicates A) 5' deoxyguanylate.
B) 3' deoxyguanylate.
C) 3', 5' deoxyguanylate.
D) 5', 3'deoxyguanylate.
E) dGMP
3. Much of the stability of the double stranded helical DNA structure is the result of
A) hydrogen bonding between purines.
B) the phosphodiester backbone.
C) Ionic nucleobase attraction.
D) the stacking interactions between base pairs.
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4/3/2013
All five histones are rich in ________ amino acid residues whose positive charges allow binding to the sugar‐phosphate backbone of DNA.
A) lysine and alanine
B) lysine and arginine
C) leucine and alanine
D) leucine and arginine
Which does not apply to most bacterial DNA? A) Circular.
B) Relaxed. C) Not packed into nucleosomes. D) Supercoiled. Which best describes the structure of a nucleosome core particle?
A) A histone octamer with DNA threaded through its center.
B) About 50 bp of DNA associated with one histone H1 molecule.
C) One nucleosome plus one histone H1 and linker DNA.
D) A histone octamer wrapped approximately two times around with DNA.
Nucleic Acid Structure
•
•
•
•
•
•
Nucleobase
Nucleoside
Nucleotide
Nucleic acid
Chromatin
Chromosome
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4/3/2013
Base Structure
• Purines and pyrimidines
• Aromatic
• Tautomers
• H‐bonding
Nucleosides
• Ribonucleosides and deoxyribonucleoside
• Purine = osine; pyrimidine = idine (watch cytosine)
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4/3/2013
Nucleotides
• Phosphorylated on 2’, 3’, or 5’
• 5’ unless noted
• Draw these:
– dA
– ADP
– ppAp
– ApAp
• pA is normally called _______ or _______________
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4/3/2013
Polynucleotides
Phosphate diesters
polyanion
directionality
5’  3’
Abbreviation is pdApdGpdTpdC
• tetranucleotide
•
•
•
•
•
Double Stranded DNA
• Chargaff’ Rule: %A =%T and %G = %C
• (C + G) not necessarily equal to (A + T)
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4/3/2013
Complementary Base Pairs
Mismatching may occur with tautomers
H
H
H
N
N
HN
N
H
N
N
NH
N
O
Adenine tautomer
Cytosine
Antiparallel
• Inaccuracy of two‐
dimensional drawing: bases are perpendicular to the paper
• “ladder”
• H‐bonding
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4/3/2013
Helix
• Maximization of base pair stacking
• More compact
• Major and minor groove
• How do we explain major/minor grooves?
Major/Minor Groove
• Many pictures show ladder with backbone at 180o
• Actually a distorted ladder with poles closer to each other, on one side
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4/3/2013
Weak Forces Stabilize Double Helix
• Stacking interactions (vdW forces)
• Hydrogen bonding
• Hydrophobic effect
• Charge‐charge
Denaturation
•
•
•
•
Melting point
Melting curve
UV‐absorption
cooperative
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4/3/2013
A/T Rich and G/C Rich strands
• GC rich strands harder to denature due to STACKING (not H‐bonds)
• Cooperativity due to initial unstacking, which exposes bases to water, which destabilizes H‐
bonds, which leads to further denaturation
Supercoiling
• Bacterial DNA
• Closed, circular DNA
• Topology and topoisomerases
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4/3/2013
Eukaryotic DNA
• Chromatin
• 8000x packing ratio
– Nucleosomes (10x)
– 30nm chromatin fiber (4x)
– RNA/Protein scaffold holds loops (200x condensation of DNA length
Nucleosomes
• Beads on string
• Histones form octamer
• Core particle
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4/3/2013
Unpacking
• Histones serve as negative supercoiling
• Histone acyltransferases
(HATS)
• Necessary for expression
Chromosome
• Scaffold of RNA and protein
• 30nm fibers are looped many times
• Picture of histone‐
depleted chromosome: DNA strands have fallen off of scaffold
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4/3/2013
Answers
1.
2.
3.
4.
5.
6.
E
B
D
B
B
D
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