DNA SYNTHESIS (REPLICATION)

Dividing cells go through an ordered series of events called the cell cycle.
1. Mitosis is the period when two sets of chromosomes are assembled, and cell division
occurs.

2. Mitosis is followed by interphase, which has three subphases: G1, S, G2 (G= gap,
S=synthesis).
a. G1 phase, a cell prepares to initiate DNA synthesis. The chromosomes uncoil and
form euchromatin.

b. DNA synthesis (replication) occurs during S phase; the DNA content doubles. RNA
synthesis (transcription) is also at high level. When DNA synthesis is complete and
most other cell constituents have doubled, the cell proceeds into G2 phase.

c. During G2 phase, the cell synthesizes the RNA and proteins required for mitosis to
occur. Chromatin condenses to form heterochromatin and the nuclear membrane
disappears.

3. A cell that has completed interphase (G1,S, G2) is ready for another round of mitosis.

Replication. This process is the production of two double-stranded DNA molecules that are
identical in every way to the parent DNA.
CHARACTERISTICS:
1. Replication is semiconservative; each daughter DNA contains one strand of parental
DNA and one newly synthesized daughter strand.

2. Catalysis. DNA polymerase (DNAPs) catalyze DNA synthesis.

a. In prokaryotes (e.g. bacteria), DNAP III is involved in replication.
b. In eukaryotes, several classes of DNAPs play an important role:
α copies the lagging strand,
δ copies the leading strand,
β carries out repair,
γ carries out mitochondrial DNA replication
Replication is a five-step process, each step involving one or more proteins and enzymes.
1. DNA-unwinding proteins or helicases unwind the DNA duplex.

Topoisomerases (in Escherichia coli, DNA gyrase) relieve the strain imposed by the
unwinding.
 2. Primase make RNA primers (short pieces of nucleic acid) that are complementary to the
DNA template strand. This process moves in the 5’-to-3’ direction on the newly
synthesized primer.
3. At the 3’ end of the primer, DNAP adds nucleotides to the 3’-OH, and the so-called
“leading strand” grows continuously in the 5’-to-3’ direction.
a. The ability of DNAPs to add incoming nucleotides only to the 3’-OH makes DNA
replication discontinuous in the other daughter strand, known as “lagging strand”.
b. The segments of newly synthesized DNA, known as Okazaki fragments, are then
linked to form a continuous DNA chain.
4. The DNAP complex removes and replace the RNA primers.
In E. coli, DNAP I, which has both 5’-to-3’ exonuclease and polymerase activities,
performs this function.
5. DNA ligase join the ends together.

Errors DNAPs are also associated with 3’-to-5’ exonuclease activity, which allows detection and
removal of mismatched base pairs. This corrective process is called editing.

DNA repair. UV irradiation, heat, pH extremes, and certain chemicals can alter purines and
pyrimidines. A variety of mechanisms are involved in the repair of damaged DNA.
1. Excision repair is a mechanism for correcting thymine dimers in DNA that are
created by UV radiation.
a. A UV-specific endonuclease “nicks”/separates one strand of the double helix,
which opens a phosphodiester bond on the 3’ side of the thymine dimer.
b. DNAP synthesizes a new DNA strand. The exonuclease activity of DNAP carries
out a 5’-3’ excision of the damaged strand.
c. DNA ligase joins the ends together.
d.
 2. Disease resulting from defective excision repair.

Xeroderma pigmentosum, a skin disease, is caused by a defective excision repair due

to a mutant UV-specific endonuclease. This condition eventually leads to skin cancer.

Ataxia-telangiectasia. Fanconi’s syndrome (anemia), and Bloom syndrome are other

diseases associated with defects in excision repair.

TEST III. DNA SYNTHESIS (REPLICATION)

DNA SYNTHESIS (REPLICATION)

1. Dividing cells go through an ordered series of events called ________ .

2. ______ is the period when __ set/s of chromosomes are assembled and cell division
occurs.

3. Mitosis is followed by _________, which has three subphases:

a. __
b. __
c. __
4. __ phase, a cell prepares to initiate DNA synthesis. The chromosomes uncoil and form
___________.

5. DNA synthesis (replication) occurs during ______; the DNA content _____. RNA
synthesis (transcription) is also at high level. When DNA synthesis is complete and most
other cell constituents have ______, the cell proceeds into ______.

6. During G2 phase, the cell synthesizes the ___and _____required for mitosis to occur.

7. ______condenses to form _______and the nuclear membrane ___________.

8. A cell that has completed interphase (G1, S, G2) is ready for another round of mitosis.

9. ______. This process is the production of ________________ that are ______ in

every way to the parent DNA.

10. Replication is _________; each daughter DNA contains one strand of parental DNA
and one newly synthesized daughter strand.

11. Catalysis. ___________ catalyze DNA synthesis.

12. In prokaryotes (e.g. bacteria), ______ is involved in replication.

13. In eukaryotes, several classes of DNAPs play an important role:

_ copies carry out mitochondrial DNA replication
_ copies the leading strand,
_ copies the lagging strand,
_ copies carry out repair

14. Replication is a ___ step process, each step involving one or more _____ and ______.

15. _______ proteins or ______ unwind the DNA duplex.

16. ____________ (in Escherichia coli, DNA gyrase) relieve the ______ imposed by the
unwinding.

17. _______ make ___ primers (short pieces of nucleic acid) that are complementary to
the DNA template strand. This process moves in the _____ direction on the newly
synthesized primer.

18. At the 3’ end of the primer, ____ adds _________ to the 3’-OH, and the so-called
“leading strand” grows continuously in the _________.

19. The ability of DNAPs to add incoming nucleotides only to the 3’-OH makes DNA
replication ___________ in the other daughter strand, known as “________”.
20. The segments of newly synthesized DNA, known as _________, are then linked to form
a continuous DNA chain.

21. The DNAP complex removes and replace the ________.

22. In E. coli, DNAP __ , which has both 5’-to-3’ exonuclease and polymerase activities,
performs this function.

23. _____ join the ends together.

24. Errors DNAPs are also associated with 3’-to-5’ ____________, which allows detection
and removal of ______________. This corrective process is called _____.

25. ________. UV irradiation, heat, pH extremes, and certain chemicals can alter purines
and pyrimidines. A variety of mechanisms are involved in the repair of damaged DNA.

26. ________ is a mechanism for _______ in DNA that are created by UV radiation.

27.
a. A UV-specific ______ ___ one strand of the _______, which opens a
_________ on the 3’ side of the thymine dimer.
b. _____ synthesizes a new _____. The exonuclease activity of DNAP carries
out a _____ of the damaged strand.
c. ______ joins the ends together.

28. Disease resulting from _____ excision repair.

29. _______________, a skin disease, is caused by a defective excision repair due to a

______ UV-specific endonuclease. This condition eventually leads to skin cancer.

30. ____________. __________ syndrome (anemia), and ____ syndrome are other
diseases associated with defects in excision repair.