Scientific History
! The march to understanding that DNA is
the genetic material
"T.H. Morgan (1908)
"Frederick Griffith (1928)
"Avery, McCarty & MacLeod (1944)
DNA
"Hershey & Chase (1952)
The Genetic Material
"Watson & Crick (1953)
"Meselson & Stahl (1958)
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1908 | 1933 1928
Genes are on chromosomes The “Transforming Factor”
! T.H. Morgan ! Frederick Griffith
"working with Drosophila " Streptococcus pneumonia bacteria
(fruit flies) ! was working to find cure for
"genes are on chromosomes pneumonia
"but is it the protein or the "harmless live bacteria mixed with
DNA of the chromosomes heat-killed infectious bacteria
that are the genes? causes disease in mice
! through 1940 proteins "substance passed from dead
were thought to be bacteria to live bacteria =
genetic material… Why? “Transforming Factor”
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1944
The “Transforming Factor” mix heat-killed
DNA is the “Transforming Factor”
pathogenic &
live pathogenic
strain of bacteria
live non-pathogenic
strain of bacteria
heat-killed
pathogenic bacteria
non-pathogenic
bacteria
! Avery, McCarty & MacLeod
A. B. C. D. " purified both DNA & proteins from
Streptococcus pneumonia bacteria
! which will transform non-pathogenic bacteria?
" injected protein into bacteria
! no effect
" injected DNA into bacteria
! transformed harmless bacteria
into virulent bacteria
mice die mice live mice live mice die
Transformation?
something in heat-killed bacteria could still transmit
disease-causing
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1952 | 1969
Confirmation of DNA Hershey Protein coat
DNA labeled
labeled
with 32P
! Hershey & Chase & Chase with 35S
T2 bacteriophages
are labeled with
"classic “blender” experiment radioactive isotopes
S vs. P
"worked with bacteriophage bacteriophages infect
bacterial cells
! viruses that infect bacteria
" grew phage viruses in 2 media, radioactively
labeled with either
bacterial cells are agitated
! 35S in their proteins to remove viral protein coats
! 32P in their DNA Which radioactive
marker is found
" infected bacteria with inside the cell?
labeled phages Which molecule
carries viral 35Sradioactivity 32P radioactivity found
genetic info? found in the medium in the bacterial cells
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Blender experiment
! Radioactive phage & bacteria in blender
"35 S phage
! radioactive proteins stayed in supernatant
! therefore protein did NOT enter bacteria
"32 P phage
! radioactive DNA stayed in pellet
! therefore DNA did enter bacteria
" Confirmed DNA is “transforming factor”
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1947 1953 | 1962
Chargaff Structure of DNA
! DNA composition: “Chargaff’s rules” ! Watson & Crick
"varies from species to species " developed double helix model of DNA
"all 4 bases not in equal quantity ! other scientists working on question:
"bases present in characteristic ratio #Rosalind Franklin
! humans: #Maurice Wilkins
A = 30.9% #Linus Pauling
T = 29.4%
G = 19.9%
C = 19.8%
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1953 article in Nature
Watson and Crick Rosalind Franklin (1920-1958)
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Double helix structure of DNA Directionality of DNA
! You need to PO4 nucleotide
number the
carbons!
" it matters! N base
5ʹ CH2
O
4ʹ ribose 1ʹ
the structure of DNA suggested a mechanism for how 3ʹ
DNA is copied by the cell 2ʹ
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OH
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5ʹ
The DNA backbone PO4
Anti-parallel strands
! Putting the DNA ! Phosphate to sugar bond
backbone together CH2
base involves carbons in 3ʹ & 5ʹ
" refer to the 3ʹ and 5ʹ ends O positions
of the DNA C "DNA molecule has
! the last trailing carbon
O
“direction”
–O P O "complementary strand runs
O
base
in opposite direction
CH2
O
“It has not escaped our notice that the specific
pairing we have postulated immediately
OH suggests a possible copying mechanism for
3ʹ the genetic material.”
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Bonding in DNA Base pairing in DNA
! Purines
5’
hydrogen "adenine (A)
bonds 3’
"guanine (G)
! Pyrimidines
phosphodiester "thymine (T)
bonds "cytosine (C)
! Pairing
3’ "A:T
5’
"C : G
….strong or weak bonds?
How do the bonds fit the mechanism for copying DNA?
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Structure of chromosomes: DNA coiling Structure of chromosomes: DNA coiling
Nucleosomes
histone core
30 nm
700 nm
diameter
Russell iGenetics Fig. 2.24
DNA vs RNA
DNA RNA
Number of strands double single
sugar deoxyribose (C5H10O4) ribose (C5H10O5)
nitrogen bases A::T, G::C A, U, C, G
Base pairing yes no
nucleus nucleus
Location in the cell mitochondrion cytoplasm
chloroplasts ribosomes
ribosomal
types --- RNA(rRNA)transfer RNA
(tRNA) messenger RNA
(mRNA)
23
Any Questions??
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