REPLICATION OF DNA
● DNA (Deoxyribonucleic Acid)
- is the genetic material of all organisms on Earth from microbes to plants and human
beings
- An organism’s complete set of DNAs, including all its genes is called genome.
- The DNA is a thin long molecule found in the cell’s nucleus which is made up of
nucleotides.
- The basic structure of nucleotide consists of a phosphate group, sugar and a
nitrogenous base
● The four different type of nucleotides of DNA
- are adenine, thymine, guanine, and cytosine which are represented by their first
letter A, T, G, C. These four nucleotides are paired as (Adenine-Thymine) and
(Guanine-Cytosine)
● DNA Replication
- is the process of DNA duplication from an existing DNA. The replication of DNA is
important for the growth repair and reproduction of cells of an organism.
- This process occurs in the nucleus of eukaryotic cells BEFORE a cell divides by
mitosis of meiosis. When a cell divides, each resulting cell keeps a copy of all your
chromosomes.
● The Major key players in DNA replication
- Helicase is the unzipping enzyme and unzips the two strands of DNA in the double
helix through the hydrogen bond that holds the two base pairs together.
- Primase will initialize the process and directs the DNA polymerase for it to figure out
where it gets to start. This primer is the starting point for DNA synthesis. The primers are
made of RNA (Ribonucleic Acid). Its major role is to act as a messenger
carrying instructions from DNA for controlling the synthesis of proteins.
- DNA polymerase is the builder enzyme which replicates DNA molecules in order to
build a new strand of DNA.
- Ligase is the gluer. Which helps glue DNA fragments together to form the new strand of
DNA.
DNA REPLICATION
Transcription
- the process where RNA is made from the DNA by copying the base sequence of the
double stranded DNA into a piece of a single stranded nucleic acid.
- This transcription process is catalyzed by the enzyme RNA Polymerase.
- Transcription of DNA to form RNA takes place in the cell’s nucleus. This process uses DNA
as a model to make an RNA (mRNA) molecule. During transcription, a strand of mRNA is
made that corresponds to a strand of DNA. Just like DNA replication, transcription also
occurs in three major steps: initiation, elongation, and termination.
DNA to RNA or mRNA
( Transcription)
● Initiation
- is the start of transcription. It transpires when the enzyme RNA polymerase binds to a
specific region of a gene which is called the promoter with the help of proteins called
‘transcription factors’. This signals the DNA double strand to unwind and open so the RNA
polymerase enzyme can ‘‘read’’ the bases found in one of the DNA strands. With the open
strands, one is considered as the template strand (anti-sense strand) and this will be used to
generate the mRNA. The other is called the non-template strand (sense strand). After
reading the bases, the RNA polymerase enzyme is now ready to make a strand of
mRNA with a complementary sequence of bases.
● Elongation
- is the adding of nucleotides to the mRNA strand. RNA polymerase reads the opened DNA
strand and forms the mRNA molecule with the use of complementary base pairs. There is a
short time during this process when the newly formed RNA is bound to the opened DNA.
During this process of elongation, an adenine (A) in the DNA binds to an uracil (U) in the
RNA. RNA polymerase does not need a primer during this process. It simply initiates the
mRNA synthesis from the starting point and then moves downstream reading the anti-sense
strand from 3’ to 5’ and generating the mRNA from the 5’ to 3’ end as it
goes. Unlike helicase enzyme in DNA replication, RNA polymerase zips DNA back up as it
goes keeping only 10-20 bases exposed one at a time.
● Termination
- is the last step of the transcription process. This happens when RNA polymerase enzyme
reaches a stop or termination sequence in the gene. When the stop sequence or stop codon
is reached, the enzyme detaches from the gene. The mRNA strand is now produced, and it
detaches from DNA. It carries with it the information encoded in the gene.By the end of
transcription, the DNA segment is transcribed to form the mRNA molecule. The template
strand shown below with the sequence T-A-C-T-A-G-A-G-C-A-T-T transcribes to form the
mRNA A-U-G-A U-C-U-C-G-U-A-A
● Translation
- is the final process of protein synthesis that takes place in the cytoplasm
● Amino acid
- The protein building block
- Amino acids combine through a dehydration link called a peptide bond
MUTATION
● Mutation
- is a permanent change of the nucleotide sequence of the genome of an organism, virus,
or extrachromosomal DNA or other genetic elements. It results in damage to DNA that is not
repaired or to RNA genomes (typically caused by radiation or chemical mutagens), errors in
the process of replication, or from the insertion or deletion of segments of DNA by mobile
genetic elements.
- These alterations can be caused by random mistakes in DNA replication or by
environmental influences such as UV rays and chemicals
● Genes
- are segments of DNA located on chromosomes. A gene mutation is defined as an
alteration in the sequence of nucleotides in DNA. This change can affect a single nucleotide
pair or larger gene segments of a chromosome. DNA consists of a polymer of nucleotides
joined together.
● Point mutation
is the change of a single nitrogen base in a DNA sequence. It usually the least harmful type
of DNA mutation. Codons are a sequence of three nitrogen bases in a row that are "read"
by messenger RNA during transcription. That messenger RNA codon is then translated
Silent Mutations: These are type of change that does not alter the sequence of a protein
because of the redundancy of the genetic code (the new triplet codes for the same amino
acid as the original triplet), or because it affects an area not coding DNA or an intron. But
this change can still have serious consequences on the phenotype. Indeed, the change of a
single nucleotide can change the splice donor site, without changing the amino acid
sequence. This may, therefore, result in a deletion of an entire exon of the peptide
sequence, the exon is not recognized because the splice site has been mutated. A
synonymous mutation means a silent mutation that affects exon, without changing the
protein sequence.
Missense Mutations: This point mutation results in the replacement of one nucleotide by
another. In some cases, this change causes a change in the amino acid encoded, which
may or may not have an impact on the function of the protein produced by the gene in the
case of a gene encoding, or the 3 affinity for a transcription factor, in the case of a promoter
region of the DNA. We speak of mutation transition when there is a substitution of a purine
base to another base purine (or pyrimidine base to another pyrimidine base). In contrast, a
mutation transversion is a mutation caused by the
replacement of a purine by a pyrimidine base (or pyrimidine base by a purine base).
Nonsense Mutation: Change of a nucleotide causes the replacement of a codon specifying
an amino acid by a stop codon. This results in the production of a truncated protein.
Frameshift Mutations
are generally much more serious and often more deadly than point mutations.
Even though only a single nitrogen base is affected, as with point mutations, in this instance,
the single base is either completely deleted or an extra one is inserted into the middle of the
DNA sequence
Insertions
add one or more extra nucleotides into the DNA. They are usually caused by transposable
elements, or errors during the replication of repeating elements
Deletions mean removing one or more nucleotides from the DNA. Like insertions, these
mutations can alter the reading frame of the gene.
a. Translocation
The joining of a fragmented chromosome to a non-homologous chromosome is a
translocation. The
piece of chromosome detaches from one chromosome and moves to a new position on
another
chromosome.
b. Deletion
This mutation results from the breakage of a chromosome in which the genetic material
becomes lost
during cell division. The genetic material can break off from anywhere on the chromosome.
c. Duplication
Duplications are produced when extra copies of genes are generated on a chromosome.
d. Inversion
In an inversion, the broken chromosome segment is reversed and inserted back into the
chromosome. If the inversion encompasses the centromere of the chromosome, it is called a
pericentric
inversion. If it involves the long or short arm of the chromosome and does not include the
centromere, it is
called a paracentric inversion
(DISEASES AND ABNORMALITIES (T^T)
1. Sickle Cell Anemia (James B. Herrick- 1910, Chicago physician)
- This genetic disease is the result of a point mutation where there is a change
in just one nucleotide in the gene for hemoglobin. The mutation causes the
hemoglobin in red blood cells to transform to a sickle shape when
de-oxygenated. Since the shape is altered, it cuts of blood circulation and
clogs the capillaries.
2. Cystic Fibrosis (Dorothy Andersen, MD- 1938)
- genetic disease that affects the secretory glands, including the mucus and
sweat glands. Cystic fibrosis causes persistent lung infections and limits the
ability to breathe over time.
3. Tay-Sachs Disease (Warren Tay, British ophthalmologist- 1881; Bernard Sachs,
American neurologist- 1896 )
- rare inherited disorder that causes progressive damage to the nervous
system and most commonly affects infants.
- caused by the absence of a vital enzyme called hexosaminidase-A (Hex-A)
- a recessively inherited disease that only occurs when both parents carry a
Tay-Sachs gene, and each parent transmits the defective gene to their child
- located on chromosome 15
4. Hemophilia (John Conrad Otto- 1803)
- inherited bleeding disorder that causes abnormal or exaggerated bleeding
and poor blood clotting
- royal disease
- passed down from parents to children, about 1/3 of cases are caused by a
spontaneous mutation, a change in a gene
- most common type of hemophilia is hemophilia A
5. Down Syndrome (John Langdon Down- 1866- British physician)
- trisomy 21 is the most common chromosomal disorder
- have 47 chromosomes in their cells instead of 46
- 3 Types: trisomy 21, mosaicism, and translocation
6. Klinefelter Syndrome (Henry Klinefelter-1940’s)
- XXY condition is a chromosomal condition that affects male physical and
cognitive development
- Most common symptoms is infertility
7. Prader Willi Syndrome (Andrea Prader and Heinrich Willi)
- a complex genetic disorder that affects growth, metabolism, appetite,
cognitive function, behavioral problems, low levels of sex hormones and a
constant feeling of hunger
- loss of genes in a specific region of chromosome 15.
8. Turner Syndrome (Henri Turner, Oklahoma physician-1938)
- associated with the x chromosome that alters development in women
- brief stature and lack of ovarian development ; infertile
9. Edward’s Syndrome (John Hilton Edwards-1960s)
- as Trisomy 18 (T18) or Trisomy E)
- all or part of an extra 18th chromosome
- 3 copies of chromosome
10. “Cri Du Chat” (Dr. Jerome Lejeune- 1963)
- ih this condition often have a high-pitched cat-like cry, small head size, and a
characteristic facial appearance.
- They may have trouble breathing and feeding difficulties. People with this condition
typically have intellectual disability, developmental and speech delay, and behavioral
issues. Cri du chat syndrome
- is due to a missing piece (deletion) of a specific part of chromosome 5 known as the
'p' arm
11. Jacobsen Syndrome (Petrea Jacobsen- 1973)
- is a condition caused by a loss of genetic material from chromosome 11.
- Because this deletion occurs at the end (terminus) of the long (q) arm of
chromosome 11, Jacobsen
- syndrome is also known as 11q terminal deletion disorder
Factors Affecting DNA Mutations
● Chemical Factors
Certain man-made chemicals have been known to cause mutations, in most cases by
revising the basic chemical composition of a cell's DNA. Ethyl methanesulfonate, a
compound used in laboratory research, affects the way that one of DNA's four component
bases behaves chemically, resulting in mutant cells with sequences of DNA different from
the parent cells. Benzopyrene, a component of cigarette smoke, and vinyl chloride, an
ingredient in plastics, affect DNA similarly.
● Radiative Factors
Our world contains different kinds of radiation, both occurring naturally and resulting from
human activity, that also encourage mutations. Ultraviolet radiation from the sun creates
bonds between bases that otherwise would not exist, causing the cell to synthesize
abnormal proteins when that section of DNA is read. Ionizing radiation, such as that emitted
as X-rays, breaks strands of DNA apart, which can lead to mutations when the cell tries to
repair its DNA using free-floating molecules.
● Biological Factors
Similar to chemicals and radiation, biological agents can cause mutations by attacking
DNA's structure. Retroviruses like HIV can insert their genetic material into a host cell's
DNA. But some viruses and bacteria also produce mutations less directly. The
long-lingering hepatitis B virus can make the body secrete defensive chemicals that, over
time, cause mutations, while the prolonged cell damage and ongoing repair resulting from
Helicobacter pylori infections may increase mutations in cells lining the stomach.
● Identifying Factors
To identify substances that may cause mutations, several biochemical tests exist, including
one that has been in use since 1973. That year, scientist Bruce Ames demonstrated that a
kind of Salmonella bacteria would grow only in the presence of mutation-causing materials.
The Ames test, in which substances are exposed to this strain of Salmonella, is still used to
identify mutagens today.
