PSYE504a

Lariosa, Bianca Isabelle

Lim, Maria Katrina BSY41
Lorino, Jessica

A. DNA POLYMERASE PROOFREADING

The most important enzyme that builds the DNA in cells are called DNA

POLYMERASES. When there is replication or copying of the deoxyribonucleic acid, these

polymerases can so-call ‘check their work’ given the bases that they themselves add. This

process is called proofreading, wherein a two nucleotides are paired together. There are instances

where these nucleotides are incorrectly paired, and so through the help of proofreading, replace

the nucleotide right away before continuing the process of DNA synthesis. Proofreading also

allows removal of unpaired 3´ overhanging nucleotides in order to create blunt ends.

Figure 1. Proofreading Process

B. MISMATCH DETECTION AND REPAIR

Although proofreading ensures the pairs of nucleotides to pair correctly, there are some

instances where some errors may occur. Mismatch repair functions to remove, as well as

replace, the pairs of nucleotides that were mismatched and slipped through during proofreading.

Mismatch repair functions by way of a protein complex initially perceiving and tying to the

mismatched base. A second complex cuts the DNA close to the mismatch, and more enzymes

moves out the wrong nucleotide and an encompassing patch of DNA. A DNA polymerase at that

point replaces the missing area with right nucleotides, and an enzyme called a DNA ligase seals

the hole.

On the other hand, mismatch detection works to correct and detect the small insertions and

deletion which occurs in the “slips” of the polymerases. The process of mismatch repair is as

follows:

1. Protein complex binds to the bases that are mismatched

2. Second complex proteins cut the DNA close to mismatch and more enzymes slices the

incorrect nucleotide as well as the surrounding patch of DNA

3. A replacement of the missing section with correct nucleotide is made possible by the

DNA Polymerases
4. DNA Ligase seals gap2 start superscript, 2 end superscript
 Figure 2. Mismatch Repair Process

Procedures that assist in repairing damaged DNA also include:

 Direct Reversal: Some DNA-damaging chemical responses can be directly "fixed" by

enzymes in the cell.

 Excision Repair: Damage to one or a couple of bases of DNA is regularly settled by

evacuation (extraction) and substitution of the damaged area. In base extraction repair,

only the damaged base is expelled. In nucleotide extraction repair, as in the mismatch

repair we saw over, a lot of nucleotides is evacuated.

 Double-stranded Break Repair: Two major pathways, non-homologous end joining and

homologous recombination, are utilized to repair double-stranded breaks in DNA (that is,

the point at which a whole chromosome parts into two pieces).

C. ABNORMALITY/ DISEASE CAUSED BY A GENETIC ERROR – PKU

 A genetic error occurs when there is a change in a person’s DNA due to an error in

replication of DNA or an environmental factor. A genetic disorder can also be caused by

mutations in the DNA of genes or changes in

the overall structure or number of

chromosomes. One specific genetic disorder

is that of Phenylketonuria—or PKU. PKU

Figure 3. The figure for Phenylalanine (symbol Phe or F[2]) is a disorder inherited from parent to

offspring that increases the levels of a

substance called phenylalanine (Phe) in the blood. Untreated PKU is the most common

biochemical cause of mental retardation. The main issue stems from the inability to convert Phe

into Tyrosine (Tyr), the precursor of dopamine. Phe levels in the bloodstream soar; Tyr levels

fall. Treatment with a diet low in Phe reduces the Phe: Tyr imbalance but cannot eliminate it.

Phe is a building block or an amino acid that is found in all proteins. If not treated, phenylalanine

can build up to harmful levels in the body, causing intellectual disability and other serious health

problems. If one has PKU, their body can't process part of Phe, which is in almost all foods.

Causes

PKU is caused by mutations in the gene that helps make an enzyme called phenylalanine

hydroxylase or PAH. This enzyme is needed to convert the amino acid phenylalanine into other

substances the body needs. When this gene, known as the PAH gene, is defective, the body

cannot break down phenylalanine. Amino acids help build protein, but phenylalanine can cause

harm when it builds up in a person's body. In particular, nerve cells in the brain are sensitive to

phenylalanine. Many different PAH mutations result in problems with breaking down
phenylalanine. Some mutations cause PKU, others cause non-PKU hyperphenylalaninemia and

others are silent mutations that do not have an effect.

PAH is particularly damaging to brain tissue where it causes brain damage. People get

phenylalanine from their diets, primarily from high protein foods, such as meat, fish, poultry,

eggs, cheese, milk, nuts, and beans. Even

breads, rice, pastas, vegetables, and fruits have

some amount of protein and contain

phenylalanine. With early recognition and

prompt treatment in a form of a low-

Figure 4. Sources of PKU and how it affects the eating habits phenylalanine diet the serious complications
and dietary nutrition of those diagnosed
of PKU can be avoided.

Furthermore, PKU is inherited, which means it is passed down through families. Both parents

must pass on a recessive copy of the gene in order for a baby to have the condition. Since infants

are missing the enzyme phenylalanine hydroxylase needed to break down the essential amino

acid phenylalanine, this buildup can harm the central nervous system and cause brain damage.

Treatment

PKU is treated by limiting the amount of protein (that contains PAH) in the diet. Treatment also

includes using special medical foods as well as special low-protein foods and taking vitamins

and minerals. People who have PKU need to follow this diet for their lifetime. It is especially

important for women who have PKU to follow the diet throughout their childbearing years. It is

vital to begin PKU treatment early and continue managing PKU for life. In the United States,

newborns are tested for PKU soon after birth as a part of the newborn screening program. PKU

experts recommend beginning management as early as possible, starting management within the
first week of life. This helps protect an infant's developing brain from the damaging effects of

high or unstable blood Phe levels. Following the strict diet can be difficult, and may lead to stress

and anxiety. If it is followed correctly there should not be any physical effects on the body.

If PKU is detected at birth, early

treatment can prevent the signs of the

Figure 5. The chemical composition of Phenylalanine and

Tyrosine, the amino acids most affected by PKU

condition mentioned in the Early Signs

section. This is why it is so important to

screen for PKU at birth. If babies start

treatment several weeks after birth,

some signs of PKU can be avoided. If treatment is started after six months of age, babies are at

risk for severe intellectual disabilities. It is important to treat PKU, even if treatment is started

after noticing signs and symptoms, in order to help prevent permanent brain damage.

Symptoms

Symptoms connected with PKU are typically absent in newly birthed infants. Affected infants

may be abnormally drowsy and unenergetic and even have difficulties in receiving milk from

their mother. In addition, untreated infants with PKU tend to have unusually light eyes, skin, and

hair, from impairment of melanin synthesis, as it is the most characteristic manifestation of PKU,

and may develop a rash that appears similar to eczema.

The signs and symptoms of PKU vary from mild to severe. The most severe form of this disorder

is known as classic PKU. Infants with classic PKU appear normal until they are a few months
old. Without treatment, these children develop permanent intellectual disability and may

encounter seizures, delayed growth, behavioral issues, and various psychiatric disorders.

Untreated individuals may have a grassy odor as a side effect of excess phenylalanine in the

body. Less severe forms of this condition, sometimes called variant PKU and non-PKU

hyperphenylalaninemia, have a lower risk of brain damage.

References

 Drugs.com. (2018). Phenylketonuria (PKU) Guide: Causes, Symptoms and Treatment

Options. Available at: https://www.drugs.com/health-guide/phenylketonuria-pku.html
 Healthline. (2018). Phenylketonuria: Causes, Symptoms, and Diagnosis. Available at:
https://www.healthline.com/health/phenylketonuria#prevention
 Khan Academy (N.D.) DNA Proofreading and Repair: Retrieved from
https://www.khanacademy.org/science/high-school-biology/hs-molecular-genetics/hs-
discovery-and-structure-of-dna/a/dna-proofreading-and-repair\
 Langman's Medical Embryology. 12th edition by Sadler, T. W. (2011)
 Mayo Clinic. (2018). Phenylketonuria (PKU) - Symptoms and causes. [online] Available
at: https://www.mayoclinic.org/diseases-conditions/phenylketonuria/symptoms-
causes/syc-20376302
 New England BioLabs Inc (2018) DNA Polymerase Proofreading. Retrieved from:
https://international.neb.com/products/pcr-qpcr-and-amplification-technologies/pcr-qpcr-
and-amplification-technologies/dna-polymerase-proofreading
 nhs.uk. (2018). Phenylketonuria. Available at:
https://www.nhs.uk/conditions/phenylketonuria/
 YourGenome (2014) What is Genetic Disorder. Retrieved from:
https://www.yourgenome.org/facts/what-is-a-genetic-disorder