ap biology 2006-2007 dna the genetic material genetics lecture #5
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AP Biology 2006-2007
DNAThe Genetic MaterialGenetics Lecture #5
AP Biology
Objectives Explain how the structure of DNA was
discovered. Explain how and why DNA replicates
itself.
AP Biology
Scientific History The march to understanding that DNA is
the genetic material T.H. Morgan (1908) Frederick Griffith (1928) Avery, McCarty & MacLeod (1944) Erwin Chargaff (1947) Hershey & Chase (1952) Watson & Crick (1953) Meselson & Stahl (1958)
AP Biology
Chromosomes related to phenotype T.H. Morgan
working with Drosophila fruit flies
associated phenotype with specific chromosome white-eyed male had specific
X chromosome
1908 | 1933
AP Biology
Genes are on chromosomes Morgan’s conclusions
genes are on chromosomes but is it the protein or the
DNA of the chromosomes that are the genes? initially proteins were thought
to be genetic material… Why?
1908 | 1933
What’s so impressiveabout proteins?!
AP Biology
The “Transforming Principle” 1928
Frederick Griffith Streptococcus pneumonia bacteria
was working to find cure for pneumonia
harmless live bacteria (“rough”) mixed with heat-killed pathogenic bacteria (“smooth”) causes fatal disease in mice
a substance passed from dead bacteria to live bacteria to change their phenotype “Transforming Principle”
AP Biology
The “Transforming Principle”
Transformation = change in phenotypesomething in heat-killed bacteria could still transmit disease-causing properties
live pathogenicstrain of bacteria
live non-pathogenicstrain of bacteria
mice die mice live
heat-killed pathogenic bacteria
mix heat-killed pathogenic & non-pathogenicbacteria
mice live mice die
A. B. C. D.
AP Biology
DNA is the “Transforming Principle” Avery, McCarty & MacLeod
purified both DNA & proteins separately 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
1944
What’s theconclusion?
mice die
AP BiologyOswald Avery Maclyn McCarty Colin MacLeod
Avery, McCarty & MacLeod Conclusion
first experimental evidence that DNA was the genetic material
1944 | ??!!
AP Biology
Confirmation of DNA Hershey & Chase
classic “blender” experiment worked with bacteriophage
viruses that infect bacteria grew phage viruses in 2 media,
radioactively labeled with either 35S in their proteins 32P in their DNA
infected bacteria with labeled phages
1952 | 1969Hershey
Why useSulfur
vs.Phosphorus?
AP Biology
Protein coat labeledwith 35S
DNA labeled with 32P
bacteriophages infectbacterial cells
T2 bacteriophagesare labeled with
radioactive isotopesS vs. P
bacterial cells are agitatedto remove viral protein coats
35S radioactivityfound in the medium
32P radioactivity foundin the bacterial cells
Which radioactive marker is found inside the cell?
Which molecule carries viral genetic info?
Hershey & Chase
AP Biology
AP Biology
Blender experiment Radioactive phage & bacteria in blender
35S phage radioactive proteins stayed in supernatant therefore viral protein did NOT enter bacteria
32P phage radioactive DNA stayed in pellet therefore viral DNA did enter bacteria
Confirmed DNA is “transforming factor”
Taaa-Daaa!
AP Biology
Hershey & Chase
Alfred HersheyMartha Chase
1952 | 1969Hershey
AP Biology
Chargaff DNA composition: “Chargaff’s rules”
varies from species to species all 4 bases not in equal quantity bases present in characteristic ratio
humans:
A = 30.9%
T = 29.4%
G = 19.9%
C = 19.8%
1947
That’s interesting!What do you notice?
RulesA = TC = G
AP Biology
Structure of DNA Watson & Crick
developed double helix model of DNA other leading scientists working on question:
Rosalind FranklinMaurice WilkinsLinus Pauling
1953 | 1962
Franklin Wilkins Pauling
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Watson and CrickWatson Explains Base pairing
CrickWatson
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Rosalind Franklin (1920-1958)
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But how is DNA copied? Replication of DNA
base pairing suggests that it will allow each side to serve as a template for a new strand
“It has not escaped our notice that the specific pairing we have postulated immediately suggests a possible copying mechanism for the genetic material.” — Watson & Crick
AP Biology
Models of DNA Replication Alternative models
become experimental predictions
conservative semiconservative
Can you designa nifty experiment
to verify?
dispersive
1
2
P
AP Biology
Semiconservative replication Meselson & Stahl
label “parent” nucleotides in DNA strands with heavy nitrogen = 15N
label new nucleotides with lighter isotope = 14N
“The Most Beautiful Experiment in Biology”
1958
parent replicationMake predictions…
15N parent strands
15N/15N
AP Biology
Predictions
1st round of replication
conservative
15N/15N
14N/14N
semi-conservative
15N/14N
dispersive
15N/14N
conservative
15N/15N
14N/14N
semi-conservative
15N/14N
dispersive
15N/14N
2nd round of replication
14N/14N
15N parent strands
15N/15N
1
2
P
AP Biology
Franklin Stahl
Matthew Meselson
Matthew Meselson Franklin Stahl
Meselson & Stahl
AP Biology
Scientific History March to understanding that DNA is the genetic material
T.H. Morgan (1908) genes are on chromosomes
Frederick Griffith (1928) a transforming factor can change phenotype
Avery, McCarty & MacLeod (1944) transforming factor is DNA
Erwin Chargaff (1947) Chargaff rules: A = T, C = G
Hershey & Chase (1952) confirmation that DNA is genetic material
Watson & Crick (1953) determined double helix structure of DNA
Meselson & Stahl (1958) semi-conservative replication
AP Biology
proteinRNA
The “Central Dogma”
DNAtranscription translation
replication
Flow of genetic information in a cell
AP Biology 2007-2008
DNA Replication
AP Biology
Double helix structure of DNA
“It has not escaped our notice that the specific pairing we have postulated immediately suggests a possible copying mechanism for the genetic material.” Watson & Crick
AP Biology
Directionality of DNA You need to
number the carbons! it matters!
OH
CH2
O
4
5
3 2
1
PO4
N base
ribose
nucleotide
This will beIMPORTANT!!
AP Biology
The DNA backbone Putting the DNA
backbone together refer to the 3 and 5
ends of the DNA the last trailing carbon
OH
O
3
PO4
base
CH2
O
base
OPO
C
O–O
CH2
1
2
4
5
1
2
3
3
4
5
5
Sounds trivial, but…this will be
IMPORTANT!!
AP Biology
Anti-parallel strands Nucleotides in DNA
backbone are bonded from phosphate to sugar between 3 & 5 carbons DNA molecule has
“direction” complementary strand runs
in opposite direction
3
5
5
3
AP Biology
Bonding in DNA
….strong or weak bonds?How do the bonds fit the mechanism for copying DNA?
3
5 3
5
covalentphosphodiester
bonds
hydrogenbonds
AP Biology
Base pairing in DNA Purines
adenine (A) guanine (G)
Pyrimidines thymine (T) cytosine (C)
Pairing A : T
2 bonds C : G
3 bonds
AP Biology
Copying DNA Replication of DNA
base pairing allows each strand to serve as a template for a new strand
new strand is 1/2 parent template & 1/2 new DNA semi-conservative
copy process
AP Biology
DNA Replication Large team of enzymes coordinates replication
Let’s meetthe team…
AP Biology
Replication: 1st step Unwind DNA
helicase enzyme unwinds part of DNA helix stabilized by single-stranded binding proteins
single-stranded binding proteins replication fork
helicase
I’d love to behelicase & unzip
your genes…
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DNAPolymerase III
Replication: 2nd step
But…We’re missing
something!What?
Where’s theENERGY
for the bonding!
Build daughter DNA strand add new
complementary bases DNA polymerase III
AP Biology
energy
ATPGTPTTPCTP
Energy of ReplicationWhere does energy for bonding usually come from?
ADPAMPGMPTMPCMPmodified nucleotide
energy
We comewith our own
energy!
And weleave behind a
nucleotide!
Youremember
ATP!Are there other ways
to get energyout of it?
Are thereother energynucleotides?
You bet!
AP Biology
Energy of Replication The nucleotides arrive as nucleosides
DNA bases with P–P–P P-P-P = energy for bonding
DNA bases arrive with their own energy source for bonding
bonded by enzyme: DNA polymerase III
ATP GTP TTP CTP
AP Biology
Adding bases can only add
nucleotides to 3 end of a growing DNA strand need a “starter”
nucleotide to bond to
strand only grows 53
DNAPolymerase III
DNAPolymerase III
DNAPolymerase III
DNAPolymerase III
energy
energy
energy
Replication energy
3
3
5B.Y.O. ENERGY!
The energy rulesthe process
5
AP Biology
Let’s Replicate!!!!DNA Video