Lectures 2 and 3 DNA and heredity

Lectures 2 and 3 DNA and heredity

Genes = unit of inheritance
What is the evidence that genes = DNA?

Robert Feulgen DNA dye (1914)

cellular location
variation among species
present in right amounts

Lectures
2
and
3

DNA
and
heredity
What is the evidence that Genes = DNA?
Traits can be transferred:
1928 Griffith & Streptococcus pneumoniae
nonvirulent
virulent

Virulence (trait) transferred from one organism to another

What is the chemical nature of this heritable change?

2 and
3
DNA=and
heredity
What isLectures
the evidence
that
Genes
DNA?
1944 Avery - experimental system
1. Create extract
2. Treat extract
3. Add to R strain

4. Test for virulence

5. DNAse!
Take home = Gene comprised of DNA
- concept of transformation
- traits can be transferred via DNA (transgenics)
DNA fulfills key properties of hereditary material

faithful duplication
informational content
variation

How?

Lectures 2 and 3 DNA and heredity

What is DNA - deoxyribonucleic acid

sugar

(charge?)

single ring

double ring

Chargaffs rules

1. Pyrimidine content = purine content

2. T = A; C=G
3. applies to dsDNA (ssDNA, ssRNA, dsRNA?)

Lectures 2 and 3 DNA and heredity

Heredity genes = DNA
Heredity requires DNA to be passed on from
generation to generation.

cytokinesis karyokinesis

Requires

replication
segregation (mitosis/meiosis)
linked to cell cycle

Lectures
and 3 DNA
and
What are the
molecular 2
underpinnings
of heredity?

heredity

Faithful replication - molecular mechanism?

How is DNA propagated?
antiparallel double stranded helix
hydrogen bonded base pairs
replication mechanism: strand separation

1st

2nd
Semiconservative
replication

Lectures 2 and 3 DNA and heredity

How is DNA propagated ?

Semiconservative
replication

Growing strand
5 end

Template strand
3 end

3 end
incoming nt
complementarity

phosphodiester linkage
phosphate ions

5 end

Lectures 2 and 3 DNA and heredity

How is DNA propagated ?

Semiconservative
replication

Prokaryotic
Circular chromosome
- single origin

Eukaryotic
linear chromosome
- multiple origins

Circular chromosome
- resolution

linear chromosome
- no resolution

Topoisomerase
- telomere??
Rate of replication?
Size of Genome?

1000nts/sec
4.7Mbs
E. coli

vs. 3.4Gbs
H. sapiens

Length of time to replicate genome?

- 9.5 yrs to read genome (Hs)
- 20-40 minutes replicate prokaryotic
- Hs??? 50bp/sec, many oris

Lectures
2
and
3

DNA
and
heredity
How is DNA propagated ?
Semiconservative
replication

Leading strand template

1. Helicase (H bonds)

3
Leading strand 5

Parent DNA

Lagging strand
3
5
Lagging strand template

Okazaki
fragment

RNA
primer

2. Primase

3. DNA Polymerase (III) + clamp (PCNA)

4. DNA Polymerase (I)
5. DNA Ligase
TTAGGG (2500X)
Telomere shortening!
Telomerase:
RNA + protein

No Telomerase = 50-200bp/division
Cancer cells telomerase!
Stem Cells telomerase!

3
5

Lectures 2 and 3 DNA and heredity

How is DNA propagated?
1. Replication
2. Segregation
3. Cytokinesis

Prokaryotic
Binary Fission
Reproduction

Eukaryotic
Mitosis
Meiosis
Reproduction
Growth
Regeneration/Repair

Lectures 2 and 3 DNA and heredity

How is DNA propagated?
1. Replication
2. Segregation
3. Cytokinesis

Importance of cytoskeleton!

Lectures
2 ?and
How is DNA
propagated

3 DNA and heredity

Fidelity of replication: if 1/105 = 60,000 mutations/division

How prevent?
1. Proofreading DNA Pol
2. Mismatch repair
3. Excision repair (ie. UV = Py dimers)
- mutations
- increased incidence of cancer
- <40% survive past 20yrs old
Biotechnology
How might knowledge of replication be useful?
3
Leading strand 5

Parent DNA

Lagging strand 3
5

3
5

DNA sequencing
Polymerase Chain Reaction (amplify DNA)

Biotechnology
Lecture
3

DNA
and heredity
How might knowledge of replication be useful?
DNA sequencing

Polymerase Chain Reaction (amplify DNA)

1. template
2. primer
3. polymerase (thermostable - Archaea)
4. ???

Applications