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DNA: Structure, Dynamics DNA: Structure, Dynamics and Recognitionand Recognition
Les Houches 2004
Richard Lavery
Institut de Biologie Physico-Chimique, Paris
Les Houches 2004
L1: Biological context, history, basic DNA structure
L2: Introductory DNA biophysics and biology
L3: DNA dynamics
L4: DNA deformation
L5: Recognizing DNA
DNA: Structure, Dynamics DNA: Structure, Dynamics and Recognitionand Recognition
Les Houches 2004
L1: Biological context, history, basic DNA structure
BIOLOGICAL CONTEXT
140 Mb
3300 Mb
0.6 Mb
4.4 Mb
4.6 Mb
VEVRLREDPETFLVQLYQHCPPLARIDSVEREPFIWSQLPTEFTIRQSTGGTMNTQIVP FT
DAATCPACLAEMNTPGERRYRYPFINCTHCGPRFTIIRAMPYDRPFTVMAAFPLCPACD FT
KEYRDPLDRRFHAQPVACPECGPHLEWVSHGEHAEQEAALQAAIAQLKMGKIVAIKGIG FT
GFHLACDARNSNAVATLRARKHRPAKPLAVMLPVADGLPDAARQLLTTPAAPIVLVDKK FT
YVPELCDDIAPDLNEVGVMLPANPLQHLLLQELQCPLVMTSGNLSGKPPAISNEQALAD FT
LQGIADGFLIHNRDIVQRMDDSVVRESGEMLRRSRGYVPDALALPPGFKNVPPVLCLGA FT
DLKNTFCLVRGEQAVLSQHLGDLSDDGIQMQWREALRLMQNIYDFTPQYVVHDAHPGYV FT
SSQWAREMNLPTQTVLHHHAHAAACLAEHQWPLDGGDVIALTLDGIGMGENGALWGGEC FT
LRVNYRECEHLGGLPAVALPGGDLAAKQPWRNLLAQCLRFVPEWQNYSETASVQQQNWS FT
VLARAIERGINAPLASSCGRFFDAVAAALGCAPATLSYEGEAACALEALAASCHGVTHP FT
VTMPRVDNQLDLATFWQQWLNWQAPVNQRAWAFHDALAQGFAALMREQATMRGITTLVF
Escherichia Coli, ≈4.6 Mb
50 m
E. coli membrane region© David S. Goodsell
500 Å
• DNA
• Double helix
• Stores genetic code as a linear sequence of bases
• ≈ 20 Å in diameter
• Human genome ≈ 3.3 x 109 bp
• ≈ 25,000 genes
Biological length scale
Chemical bond 1 Å (10-10 m)
Amino acid 10 Å (10-9 m)
Globular protein 100 Å (10-8 m)
Virus 1000 Å (10-7 m)
Cell nucleus 1 m (10-6 m)
Bacterial cell 5 m (10-5 m)
Chromosome DNA 10 cm (10-1 m)
Biological length scale
If 20 Å 1 cm then ...
1 m 5000 km ...
Nucleus 15 m2 room
A "minimal" organism
"We are wondering if we can come up with a molecular definition of life"
"The goal is to fundamentally understand the components of the most basic living cell"
Craig Venter, founder of Celera Genomics, IBEA and several other gene tech companies
Hutchinson et al. Science 286, 1999, 2165
Modelling the budding yeast cell cycle (Tyson & Novak)
K.C. Chen et al. Mol. Biol. Cell Cycle 11 (2000) 369
E-cell project
580,000 bp
500 genes
Molecular machines .... transcriptosome
Nanobiotechnology D.S. Goodsell
Nucleosome
A LITTLE HISTORY ...
History of DNA
1865 Gregor Mendel publishes his work on plant breeding with the notionof "genes" carrying transmissible characteristics
1869 "Nuclein" is isolated by Johann Friedrich Miescher à Tübingen
in the laboratory of Hoppe-Seyler 1892 Meischer writes to his uncle "large biological molecules composed
of small repeated chemical pieces could express a rich language inthe same way as the letters of our alphabet"
1920 Recognition of the chemical difference between DNA and RNA
Phoebus Levene proposes the "tetranucleotide hypothesis" 1938 William Astbury obtains the first diffraction patters of DNA fibres
History of DNA
1944 Oswald Avery (Rockefeller Institute) proves that DNA carries thegenetic message by transforming bacteria
History of DNA
1950 Erwin Chargaff discovers A/G = T/C
History of DNA
1953 Watson and Crick propose the double helix as the structure of DNAbased on the work of Erwin Chargaff, Jerry Donohue, Rosy Franklinand John Kendrew
Maurice Wilkins – Kings College, London
Rosalind Franklin (in Paris)
X-ray fibre diffraction pattern of B-DNA
Linus Pauling’s DNA
Watson-Crick base pairs
Thymine -Adenine Cytosine -Guanine
Watson and Crick
It has not escaped our notice …
It has not escaped our notice that the specific pairing we have postulated suggests a possible copying mechanism for the genetic material.
Double helix ?
Dickerson Dodecamer (Oct. 1980)
DNA STRUCTURE
Nucleoside
Nucleotide
OH ribose
H deoxyribose
.
O
H
H
OH
H
H
H
O N
N
NH2
CH2P O
O
O -
P O
O
O -
P O- O
O
O - 1'
2'3'
4'
5'
2'-deoxycytidine 5' triphosphate
O
H
H
H
H
CH2P O
O
O -
P O
O
O -
P O- O
O
O - 1'
2'3'
4'
5'
N
N
NH2
N
N
2'-deoxyadenosine 5' triphosphate
OH H
O
H
H
OH
H
H
H
CH2P O
O
O -
P O
O
O -
P O- O
O
O - 1'
2'3'
4'
5'
N
NHN
N
O
H2N
2'-deoxyguanosine 5' triphosphate
O
H
H
OH
H
H
H
CH2P O
O
O -
P O
O
O -
P O- O
O
O - 1'
2'3'
4'
5'
O N
H N
O
CH3
thymidine 5' triphosphate
Nucleotide triphosphates
Nucleotides are linked by phosphodiester bonds
Strand has a direction (5'3')
DNA/RNA chemical structure
DNA : A ,T,G,C + deoxyriboseRNA : A,U,G,C + ribose
Base families
Purine (Pur / R) Pyrimidine (Pyr / Y)
C2
N1
C5C6N7
C4
C8
N9
N3
N1
C4
N3
C2
C5
C6
Watson-Crick base pairs
Thymine -Adenine Cytosine -Guanine
Base pair dimensions
34 Å
3.4 Å
20 Å
MinorGroove
MajorGroove
GC
CG
AT
TA
CG
GCAT
TA
TA
AT
GCCG
GC
Strands areantiparallel
CGCGTTGACAACTGCAGAATC
A and B DNA allomorphs
B A
Hydration
Antiparallel strands
5’5’3’
3’
DNA grooves
MINOR
MAJOR
B-DNA (longitudinal view)
B-DNA (lateral view)
R.H. helix
A-DNA (longitudinal view)
A-DNA (lateral view)
R.H. helix
Z-DNA (longitudinal view)
Z-DNA (lateral view)
L.H. helix
Base pairs are rotated in Z-DNA
Backbone dihedrals - I
0
+60°+10°
Dihedral angle definition
Staggered Eclipsed
Favoured conformations
gauche +
trans
gauche -
Backbone dihedrals - II
: O3’ – P – O5’ – C5’ g-
: P – O5’ – C5’ – C4’ t
: O5’ – C5’ – C4’ – C3’ g+
: C5’ – C4’ – C3’ – O3’ g+
: C4’ – C3’ – O3’ – P t
: C3’ – O3’ – P – O5’ g-(Y) : O4’ – C1’ – N1 – C2 g-
(R) : O4’ – C1’ – N9 – C4
syn-anti glycosidic conformations
Baird & Tatlock 1901
Medicine SetsManufactures by Messrs Burroughs, Wellcome & Co.
Sugar ring puckering
C5’
ENDO
EXO
Base
Sugar pucker described as
pseudorotation
North : C3’-endo
East : O4’-endo
South : C3’-endo
"2 B or not 2 B ...." W. Shakespeare 1601
Pseudorotation Equations
Altona et al. J. Am. Chem. Soc. 94, 1972, 8205
0
2
13
4
Basetan P = (4 - 1) - (3 - 0)
22 (Sin 36° + Sin72°)
Amp = 2 / Cos P
Preferred sugar puckers
Sugar pucker and P-P distance
UNUSUAL DNA STRUCTURES
Alternative base pairs
Watson-Crick
Reversed Watson-Crick
Hoogsteen Reversed Hoogsteen
Watson-Crick + Hoogsteen = Base triplet
- note C(N3) protonation
Triple helix DNA
Guanine Hoogsteen pairing Base tetraplex
Quadruplex DNA
Inverted repeat can lead to loop formation
DNA cruciform
Holliday junction
PNA versus DNA
Peptide Nucleic acid(PNA)
Achiral, peptide-like backbone
Backbone is uncharged High thermal stability
High-specificity hybridization with DNA
Resistant to enzymatic degradation
Can displace DNA strand of duplex
Pyrimidine PNA strands can form 2:1 triplexes with ssDNA
Biotechnological applications
Parallel-stranded DNA
I-DNA: intercalated parallel-stranded duplexes
and nucleotide anomers
H OH is not the only change in passing from DNA to RNA ....
Books on DNA
Principles of Nucleic Acid Structure, W. Saenger, 1984 Springer-Verlag
Nucleic Acid Structure, Ed. S. Neidle, 1999 Oxford University Press
DNA Structure and Function, R.R. Sinden, 1994 Academic Press
Biochemistry, D. Voet and J.G. Voet, 1998 DeBoeck
The Eighth Day of Creation, H.F. Judson, 1996 Cold Spring Harbour Press