Biochemistry

Tutor 8
Role of DNA as genetic
information
 First extracted from nuclei
in 1870
deoxyribose nuclei acid
Chemical analysis: DNA
was a weak acid rich in
phosphorous
contain sugar moiety

DNA Characteristic:
(deoxyribose)

weakly acidic

found in nucleus
 Chromosome also contain PROTEIN, so
early it was a challenge to demonstrate
that DNA indeed the molecule that
contained genetic information
Are genes
composed of Chromosome contain more PROTEIN
than DNA
DNA or Varitaions:
PROTEIN? - DNA has 4 different subunits
- PROTEIN has 20 different
subunits
Fredrick Griffith “Transforming Principle”
Blood of the mice on fourth experiment:
Find living bacteria inside, and somehow the R
bacteria had transformed into S bacteria

Conclusion:
there was a “transforming principle” responsible for
this

Further question:
Was it proteins in the bacteria, sugar coat on S
bacteria, immune system of the mouse, or nucleic
acids (RNA and DNA)?
Oswald-Avery Project finding the
“Trnasforming Principle” Component
Conclusion:
When the DNA destroyed, there are no
transformation occurred

So the “transforming principle”

component must be DNA!
The Hershey-Chase
Experiment
Conclusion:
DNA carried the instructions to make new
viruses, which was passed to subsequent
generations.
Chargaff and his “rules”

Erwin Chargaff helped pave the way to understanding the three-dimensional

structure of the DNA molecule.

Chargaff’s Rules’:
1. In DNA, regardless of which organism it comes from, the amount of adenine (A) is usually the same as the amount
of thymine (T), and the amount of guanine (G) is usually the same as the amount of cytosine (C).

2. The composition of DNA varies between different species such that the amount of each base is different. This
diversity in the composition of DNA made it a much more credible candidate for the genetic material than protein.
Regulation of Protein
Synthesis:
DNA replication and
transcription
 DNA
transcribes into
RNA
which then get translated into
PROTEIN
Complex interplay of many macromolecules
1. Ribosomes: human ribosomes has 2 subunit (small 40S and large 60S). Functional sites  A (aminoacyl), P (Peptidyl),

E (Exit).

2. mRNA: carry a coding section of a gene for protein synthesis.  codon

3. tRNA: adaptors bridging mRNA to amino acid sequence found in a growing protein.  anti-codon

4. Genetic code: three nucleotides encoded genome that specifies individual amino acids found in proteins. 64 sequence (3

stop codons) used to specify 20 amino acids.

5. Protein factors: non-ribosomal proteins that participate during initiation, elongation, and termination process.
 Initiation

• Identifying appropriate codon to initiates transcription

(promoter)
• Promoter as a site for RNA polymerase to start encoding
• Double helix DNA split by helicase enzyme

Transcriptio
n (in Elongation

• DNA leading strand or anti-sense (3’-5’) translated by

nucleus) RNA polymerase into RNA that reflecting lagging strand
or sense (5’-3’)

Termination

• Transcription process is done at terminator region, DNA

and the new RNA polymerase split up
Transcription
Splicing
 Translation (in cytoplasma and ribosome)

Base in mRNA grouped into triplets  codon

Start codon (AUG), stop codon (UGA, UAG, UAA)

Small ribosome subunit + mRNA from start codon

tRNA with AA and anti-codon binds at start codon

Large ribosome unit + mRNA + tRNA+ small

ribosome unit make up the translation complex and
work together to form polypeptide

The process stops when stop codone at P functional

site + release factor at A functional site  polypeptide
released
Genetic Code and Human Disease
Appropriate readout of the genetic code is essential for human health. Mutations that alter protein-coding
sequences can affect proteins in many different ways. The effect of mutations on the coding sequence can
classify as either

1. Synonymous: Changes in base not affect protein sequence

2. Nonsynonymous: affecting protein sequence

a. Missense: base code changes that results in amino acid changes

b. Nonsense: Change into stop codon  premature termination  shorter protein sequence

c. Frameshift: insertion or deletion of base results in shifting the reading frame

 Many human diseases result from changes in protein sequence caused by mutations that
alter the correct readout of genetic information from gene to a functional protein. Defects in
the protein synthetic machinery also cause a small but growing number of human diseases.

Those mutations in human genetic can be passed genotypically to generations including

mutations that occur in BREAST CANCER.
 Genetic material
knowledge to genetic
counseling in cancer
HBOC Syndrome
BRCA1 and BRCA2

 The most common cause of hereditary breast cancer is an inherited mutation in

the BRCA1 or BRCA2 gene. In normal cells, these genes help make proteins that repair
damaged DNA. Mutated versions of these genes can lead to abnormal cell growth, which can
lead to cancer.

• Women with a BRCA gene change have a greatly increased risk of getting breast cancer, as well as an increased risk of
ovarian cancer, pancreatic cancer, and possibly some other cancers. 

• Men with a BRCA gene change are at increased risk of breast cancer (although this risk is lower than in women to
begin with), prostate cancer, pancreatic cancer, and possibly some other cancers. 
Higher Risk based on:
• Closeness of affected family members
• Numbers of family members affected
• Age when the relatives were diagnosed
• Common in jewish people of Ashkenazi
• Risk having cancer in both breasts and younger age
BRCA1

As an autosomal dominant
syndrome, a deleterious
BRCA1 mutation can be
transmitted through maternal or
paternal lineages
BRCA2

As an autosomal dominant
syndrome, a deleterious BRCA2
mutation can be transmitted through
maternal or paternal lineages
Genetic Counseling
Risk assessment tools  with genetic counselor  review family history  weight the risks and
benefits of getting the genetic testing

Women who have already been diagnosed with Other groups of people: 

breast cancer:  • People with a known family history of a BRCA mutation

• You were diagnosed with breast cancer at a younger

• Women diagnosed with ovarian cancer or pancreatic cancer, or men
age
diagnosed with breast cancer, pancreatic cancer, or high-grade or
• You have been diagnosed with breast cancer a second metastatic prostate cancer
time (not a recurrence of the first cancer)
• People with a family history of breast cancer at a younger age, more
• You are of Ashkenazi Jewish descent than one family member with breast cancer, or breast cancer in a

• You have a family history of breast cancer, ovarian male family member

cancer, pancreatic cancer, or prostate cancer • People with a close family member with a history of ovarian cancer,

pancreatic cancer, or metastatic prostate cancer

Natural history of antibody
regarding the use of
monoclonal antibody
 History of Antibody Development

18th century
Edward Jenner: discovery that fluid obtained
from smallpox pustule can provide immunity
from acquiring the disease.

Gerald Eldeman & Rodney Porter:

understanding antibody structure

20th century
 1942 Bjørneboe and Gormsen:
Concluding that plasma cell were the
source of antibody production

History of
Moore, Kabat, and Gutman: studying
Monoclonal protein characteristic of myeloma
Antibody
Confirmation of their origin from a
single plasma cell clone
Concept of
Monoclonal Antibody
Fused an antibody-producing plasma cell with a
myeloma cell

 Hybridoma Cells  inherited immortality from

the myeloma cells and selective- resistance from the
primary B-lymphocytes

 produce unlimited quantities of monospecific

antibodies
References
• Campbell 2017
• https://www.yourgenome.org/stories/revealing-dna-as-the-molecule-of
-life
• http://www.csun.edu/~cmalone/pdf360/Ch10-1%20Gen%20material.p
df
• https://www.ncbi.nlm.nih.gov/books/NBK545161/
• Cancer.org
• Visualsonline.cancer.gov
• https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5357605/