DNA Repair
DNA Repair
Most mistakes during replication are corrected by DNA polymerase during replication or
by post-replication repair mechanisms.
LEARNING OBJECTIVES
Explain how errors during replication are repaired
KEY TAKEAWAYS
Key Points
Mismatch repair enzymes recognize mis-incorporated bases, remove them from
DNA, and replace them with the correct bases.
In nucleotide excision repair, enzymes remove incorrect bases with a few
surrounding bases, which are replaced with the correct bases with the help of a
DNA polymerase and the template DNA.
When replication mistakes are not corrected, they may result in mutations, which
sometimes can have serious consequences.
Point mutations, one base substituted for another, can be silent (no effect) or may
have effects ranging from mild to severe.
Mutations may also involve insertions (addition of a base), deletion (loss of a
base), or translocation (movement of a DNA section to a new location on the
same or another chromosome ).
Key Terms
mismatch repair: a system for recognizing and repairing some forms of DNA
damage and erroneous insertion, deletion, or mis-incorporation of bases that can
arise during DNA replication and recombination
nucleotide excision repair: a DNA repair mechanism that corrects damage done
by UV radiation, including thymine dimers and 6,4 photoproducts that cause bulky
distortions in the DNA
Errors during Replication
DNA replication is a highly accurate process, but mistakes can occasionally occur as
when a DNA polymerase inserts a wrong base. Uncorrected mistakes may sometimes
lead to serious consequences, such as cancer. Repair mechanisms can correct the
mistakes, but in rare cases mistakes are not corrected, leading to mutations; in other
cases, repair enzymes are themselves mutated or defective.
Mutations: In this interactive, you can “edit” a DNA strand and cause a mutation. Take
a look at the effects!
Most of the mistakes during DNA replication are promptly corrected by DNA polymerase
which proofreads the base that has just been added. In proofreading, the DNA pol reads
the newly-added base before adding the next one so a correction can be made. The
polymerase checks whether the newly-added base has paired correctly with the base in
the template strand. If it is the correct base, the next nucleotide is added. If an incorrect
base has been added, the enzyme makes a cut at the phosphodiester bond and
releases the incorrect nucleotide. This is performed by the exonuclease action of DNA
pol III. Once the incorrect nucleotide has been removed, a new one will be added again.
DNA polymerase proofreading: Proofreading by DNA polymerase corrects errors during replication.
Some errors are not corrected during replication, but are instead corrected after
replication is completed; this type of repair is known as mismatch repair. The enzymes
recognize the incorrectly-added nucleotide and excise it; this is then replaced by the
correct base. If this remains uncorrected, it may lead to more permanent damage. How
do mismatch repair enzymes recognize which of the two bases is the incorrect one?
In E. coli, after replication, the nitrogenous base adenine acquires a methyl group; the
parental DNA strand will have methyl groups, whereas the newly-synthesized strand
lacks them. Thus, DNA polymerase is able to remove the incorrectly-incorporated bases
from the newly-synthesized, non-methylated strand. In eukaryotes, the mechanism is
not very well understood, but it is believed to involve recognition of unsealed nicks in the
new strand, as well as a short-term continuing association of some of the replication
proteins with the new daughter strand after replication has been completed.
Mismatch Repair: In mismatch repair, the incorrectly-added base is detected after replication. The mismatch-
repair proteins detect this base and remove it from the newly-synthesized strand by nuclease action. The gap is
now filled with the correctly-paired base.
In another type of repair mechanism, nucleotide excision repair, enzymes replace
incorrect bases by making a cut on both the 3′ and 5′ ends of the incorrect base. The
segment of DNA is removed and replaced with the correctly-paired nucleotides by the
action of DNA pol. Once the bases are filled in, the remaining gap is sealed with a
phosphodiester linkage catalyzed by DNA ligase. This repair mechanism is often
employed when UV exposure causes the formation of pyrimidine dimers.
DNA Ligase I Repairing Chromosomal Damage: DNA damage, due to environmental factors and normal
metabolic processes inside the cell, occurs at a rate of 1,000 to 1,000,000 molecular lesions per cell per day. A
special enzyme, DNA ligase (shown here in color), encircles the double helix to repair a broken strand of DNA.
DNA ligase is responsible for repairing the millions of DNA breaks generated during the normal course of a cell’s
life. Without molecules that can mend such breaks, cells can malfunction, die, or become cancerous. DNA ligases
catalyse the crucial step of joining breaks in duplex DNA during DNA repair, replication and recombination, and
require either Adenosine triphosphate (ATP) or Nicotinamide adenine dinucleotide (NAD+) as a cofactor.
Nucleotide Excision Repairs: Nucleotide excision repairs thymine dimers. When exposed to UV, thymines lying
adjacent to each other can form thymine dimers. In normal cells, they are excised and replaced.
DNA Damage and Mutations
Errors during DNA replication are not the only reason why mutations arise in DNA.
Mutations, variations in the nucleotide sequence of a genome, can also occur because
of damage to DNA. Such mutations may be of two types: induced or spontaneous.
Induced mutations are those that result from an exposure to chemicals, UV rays, X-
rays, or some other environmental agent. Spontaneous mutations occur without any
exposure to any environmental agent; they are a result of natural reactions taking place
within the body.
Mutations may have a wide range of effects. Some mutations are not expressed; these
are known as silent mutations. Point mutations are those mutations that affect a single
base pair. The most common nucleotide mutations are substitutions, in which one base
is replaced by another. These can be of two types: transitions or transversions.
Transition substitution refers to a purine or pyrimidine being replaced by a base of the
same kind; for example, a purine such as adenine may be replaced by the purine
guanine. Transversion substitution refers to a purine being replaced by a pyrimidine or
vice versa; for example, cytosine, a pyrimidine, is replaced by adenine, a purine.
Mutations can also be the result of the addition of a base, known as an insertion, or the
removal of a base, known as a deletion. Sometimes a piece of DNA from one
chromosome may get translocated to another chromosome or to another region of the
same chromosome.
LICENSES AND ATTRIBUTIONS
