1. Nucleic Acids
Information storage

2. Nucleic Acids Function: genetic material stores information
genes
blueprint for building proteins
DNA  RNA  proteins
transfers information
blueprint for new cells
blueprint for next generation
DNA
proteins

3. Nucleic Acids Examples: Structure: RNA (ribonucleic acid)
single helix
DNA (deoxyribonucleic acid)
double helix
Structure:
monomers = nucleotides
DNA
RNA

4. Nucleotides 3 parts nitrogen base (C-N ring) pentose sugar (5C)
ribose in RNA
deoxyribose in DNA
phosphate (PO4) group
Nitrogen base I’m the A,T,C,G or U part!
Are nucleic acids charged molecules?
DNA & RNA are negatively charged:
Don’t cross membranes.
Contain DNA within nucleus
Need help transporting mRNA across nuclear envelope.
Also use this property in gel electrophoresis.

5. Types of nucleotides 2 types of nucleotides different nitrogen bases
Purine = AG
Pure silver!
2 types of nucleotides
different nitrogen bases
purines
double ring N base
adenine (A)
guanine (G)
pyrimidines
single ring N base
cytosine (C)
thymine (T)
uracil (U)

6. Dangling bases? Why is this important?
Nucleic polymer
Backbone
sugar to PO4 bond
phosphodiester bond
new base added to sugar of previous base
polymer grows in one direction
N bases hang off the sugar-phosphate backbone
Dangling bases? Why is this important?

7. Pairing of nucleotides
Nucleotides bond between DNA strands
H bonds
purine :: pyrimidine
A :: T
2 H bonds
G :: C
3 H bonds
The 2 strands are complementary.
One becomes the template of the other & each can be a template to recreate the whole molecule.
Matching bases? Why is this important?

8. H bonds? Why is this important?
DNA molecule
Double helix
H bonds between bases join the 2 strands
A :: T
C :: G
H bonds = biology’s weak bond
• easy to unzip double helix for replication and then re-zip for storage
• easy to unzip to “read” gene and then re-zip for storage
H bonds? Why is this important?

9. Matching halves? Why is this a good system?
Copying DNA
Replication
2 strands of DNA helix are complementary
have one, can build other
have one, can rebuild the whole
when cells divide, they must duplicate DNA exactly for the new “daughter” cells
Why is this a good system?
Matching halves? Why is this a good system?

10. When does a cell copy DNA?
When in the life of a cell does DNA have to be copied?
cell reproduction
mitosis
gamete production
meiosis
when cells divide, they must duplicate DNA exactly for the new “daughter” cells
Why is this a good system?

11. DNA Replication

12. Directionality of DNA You need to number the carbons! nucleotide
it matters!
nucleotide
PO4
N base 5
CH2
This will be IMPORTANT!! O 4 1
ribose 3 2 OH

13. Sounds trivial, but… this will be IMPORTANT!!5
The DNA backbone
PO4
Putting the DNA backbone together
refer to the 3 and 5 ends of the DNA
the last trailing carbon
base
CH2 5 O 4 1 C 3 2 O –O P O
Sounds trivial, but… this will be IMPORTANT!! O
base
CH2 5 O 4 1 3 2 OH 3

14. 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 5 3 3 5

15. Bonding in DNA 5 3 3 5 hydrogen bonds covalent phosphodiester
….strong or weak bonds?
How do the bonds fit the mechanism for copying DNA?

16. Base pairing in DNA Purines Pyrimidines Pairing adenine (A)
guanine (G)
Pyrimidines
thymine (T)
cytosine (C)
Pairing
A : T
2 bonds
C : G
3 bonds

17. 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

18. DNA Replication Large team of enzymes coordinates replication
Let’s meet the team…
DNA Replication
Large team of enzymes coordinates replication
Enzymes
more than a dozen enzymes & other proteins participate in DNA replication

19. Replication: 1st step Unwind DNA helicase enzyme
I’d love to be helicase & unzip your genes…
Replication: 1st step
Unwind DNA
helicase enzyme
unwinds part of DNA helix
stabilized by single-stranded binding proteins
helicase
single-stranded binding proteins
replication fork

20. Where’s the ENERGY for the bonding! We’re missing something!
Replication: 2nd step
Build daughter DNA strand
add new complementary bases
DNA polymerase III
Where’s the ENERGY for the bonding!
But…
We’re missing something!
What?
DNA
Polymerase III

21. Leading & Lagging strands
Okazaki
Leading & Lagging strands
Limits of DNA polymerase III
can only build onto 3 end of an existing DNA strand  5
Okazaki fragments 5 5 3 5 3 5 3
ligase
Lagging strand 3
growing
replication fork 3 5
Leading strand  3 5
Lagging strand
Okazaki fragments
joined by ligase
“spot welder” enzyme 3
DNA polymerase III
Leading strand
continuous synthesis

22. direction of replication
Replication fork
DNA polymerase III
lagging strand
DNA polymerase I 3’
primase
Okazaki fragments 5’ 5’
ligase
SSB 3’ 5’ 3’
helicase
DNA polymerase III 5’
leading strand 3’
direction of replication
SSB = single-stranded binding proteins

23. DNA polymerase III enzyme
DNA polymerases
DNA polymerase III
1000 bases/second!
main DNA builder
DNA polymerase I
20 bases/second
editing, repair & primer removal
DNA polymerase III enzyme
In 1953, Kornberg was appointed head of the Department of Microbiology in the Washington University School of Medicine in St. Louis. It was here that he isolated DNA polymerase I and showed that life (DNA) can be made in a test tube. In 1959, Kornberg shared the Nobel Prize for Physiology or Medicine with Severo Ochoa — Kornberg for the enzymatic synthesis of DNA, Ochoa for the enzymatic synthesis of RNA.

24. Editing & proofreading DNA
1000 bases/second = lots of typos!
DNA polymerase I
proofreads & corrects typos
repairs mismatched bases
removes abnormal bases
repairs damage throughout life
reduces error rate from 1 in 10,000 to 1 in 100 million bases