Curs 5

Replicarea celulara

Replicarea ADN
 CICLUL CELULAR Ciclul celular
consta din
interfaza (cind
celula nu se
replica) si
replicarea
celulara sau
mitoza

Mitoza este procesul de diviziune celulară, specific celulelor

eucariote, prin care dintr-o celulă-mamă rezultă doua celule-
fiice, identice din punct de vedere genetic
 Mitoza si Meioza
• Mitoza:
- diviziunea celulara a celulelor somatice (din
intregul corp) – pe baza ei se realizeaza cresterea
organismului (celulele fiice vor avea 2n cromozomi
= diploide)
• Meioza:
• – are loc la organismele cu reproducere sexuata
- diviziunea celulara a celulelor sexuale care sunt
haploide avind un singur set de cromozomi (n) -
gametii
Fazele Mitozei

• Profaza

• Metafaza

• Anafaza

• Telofaza
 Interfaza
• Etapa in care celula se pregateste de mitoza
• Alcatuita din fazele G1 (sinteza de ARN, proteine si
acumulare de ATP), S (replicarea ADN), G2 (sinteza
unor proteine necesare formării fusului de diviziune –
tubulina, actina, miozina)
1. Celula are cea mai ridicata activitate metabolica
2. Materialul genetic se dubleaza
 Profaza
1. Cromozomii devin vizibili la
microscopul optic si se dubleaza
- cromozomii dublati prezinta doua
cromatide (surori) legate prin
centromer
2. Centriolii migreaza in doua capete
opuse ale nucleului
3. Nucleolii dispar
4. Membrana nucleara se
dezintegreaza

Photographs from: http://www.bioweb.uncc.edu/biol1110/Stages.htm

 Metafaza
1. Cromozomii se dispun la “ecuatorul” celulei (in mijloc)
2. Se ataseaza cu ajutorul centromerilor de fibrele fusului
mitotic si formeaza placa metafazica (ecuatoriala)
3. La sfarsitul metafazei are loc diviziunea centromerilor,
cromozomii devenind monocromatidici
 Anafaza
1. Cromozomii se cliveaza pe axul longitudinal
rezultand doi cromozomi formati dintr-o
singura cromatida
2. Cromozomii monocromatidici vor migra pe
fibrele fusului de diviziune spre polii opusi ai
celulei
 Telofaza
1. Cromozomii se despiralizeaza
2. Fibrele fusului de diviziune se dezintegreaza
3. Se formeaza cite un nucleu cu membrana
nucleara la fiecare pol unde au migrat
cromozomii
4. Celula se divide (citokineza) – citoplasma si
organitele se divizeaza si apar cele doua celule
fiice, identice, cu garnitura diploida de
cromozomi (2n)
Mitoza
 METAPHASE ANAPHASE TELOPHASE AND CYTOKINESIS
Metaphase
plate Cleavage Nucleolus
furrow forming

Nuclear
envelope
Spindle Centrosome at Daughter forming
one spindle pole chromosomes
 Meioza
• Intr-o runda de meioza se obtin 4 fiice (gameti)
• Fiecare din cele celulele rezultate contine jumatate din
garnitura de cromozomi din celulelor parentale (sunt
haploide = n cromozomi)
• Au loc doua seturi de diviziune:
• 1. in prima faza dintr-o celula 2n se obtin doua celule n
• 2. in a doua faza (mitotica) din doua celule n se obtin 4
celule, tot cu garnitura haploida n = gametii, care nu se
mai divid
 DNA and RNA are polymers of
nucleotides

Phosphate
group

Nitrogenous
base
Sugar Nitrogenous base
(A, G, C, or T)
Phosphate
group
Nucleotide

Thymine (T)

Sugar
(deoxyribose)

DNA nucleotide
Polynucleotide Sugar-phosphate backbone
• Each strand of the 5 end 3 end

double helix is P
oriented in the
opposite direction
P
P

P P

P P

3 end 5 end
 How can entire chromosomes be replicated during S phase?

• DNA replication begins at many specific sites

Origin of replication Parental strand

Daughter strand

Bubble

Two daughter DNA molecules

 Replicarea ADN este semiconservativa

Replication of DNA:

•base pairing allows each strand

to serve as a template for a new
strand
•new strand is 1/2 parent
template and 1/2 new DNA
 Replication: 1st step
• Unwind DNA
– helicase enzyme
• unwinds part of DNA helix
• stabilized by single-stranded binding proteins
helicase

single-stranded binding proteins replication fork

 DNA Polymerase
• Bidirectional synthesis of the DNA double
helix
• Corrects mistaken base pairings
• Requires an established polymer (small RNA
primer)
primer before addition of more nucleotides
• Other proteins and enzymes necessary
 Replication: 2nd step
 Build daughter DNA
strand
 add new complementary
bases
 DNA polymerase III

DNA
Polymerase III
Energy of Replication

energy
energy

TTP
CTP
GTP
ATP TMP
AMP
CMP
GMP
ADP
modified nucleotide
 Energy of Replication
• The nucleotides arrive as nucleosides
– DNA bases with P–P–P
• P-P-P = energy for bonding
– DNA bases arrive with their own energy source for
bonding
– bonded by enzyme: DNA polymerase III

ATP GTP TTP CTP

 5 3

Replication
energy
DNA
• Adding bases Polymerase III
– can only add nucleotides energy
to DNA
Polymerase III
3 end of a growing DNA
strand energy
DNA
• need a “starter” nucleotide
Polymerase III
to
bond to
DNA
energy
– strand only grows Polymerase III
53
3 5
 Leading & Lagging strands
Limits of DNA polymerase III
 can only build onto 3 end of an
existing strand 5


r agme nts
aki f
Okaz 5
3 5 5 3
3
5 Lagging strand
3
ligase
growing 3
replication fork
5
Leading strand

Lagging strand
3
 5

3
DNA polymerase III
 Okazaki fragments
 joined by ligase Leading strand
 continuous synthesis
 DNA Replication

• Priming:
1. RNA primers:
primers before new DNA strands can
form, there must be small pre-existing
primers (RNA) present to start the addition of
new nucleotides (DNA Polymerase).
Polymerase)

2. Primase:
Primase enzyme that polymerizes
(synthesizes) the RNA Primer.
 DNA Replication

• Synthesis of the new DNA Strands:

1. DNA Polymerase:
Polymerase with a RNA primer in
place, DNA Polymerase (enzyme) catalyze
the synthesis of a new DNA strand in the 5’ to
3’ direction.
direction

5’ 3’

RNA
5’
DNA Polymerase Primer
Nucleotide
 DNA Replication

2. Leading Strand:
Strand synthesized as a
single polymer in the 5’ to 3’ direction.
direction

5’ 3’
5’
RNA
Nucleotides DNA Polymerase Primer
 DNA Replication

3. Lagging Strand:
Strand also synthesized in
the 5’ to 3’ direction,
direction but discontinuously
against overall direction of replication
(replication fork)
Leading Strand
5 3’
’
3’ 5’
DNA Polymerase RNA Primer
5’ 3’

3’ 5’
Lagging Strand
 DNA Replication

4. Okazaki Fragments:
Fragments series of short
segments on the lagging strand.

DNA
Okazaki Fragment Polymerase
RNA
Primer
5’ 3’

3’ 5’
Lagging Strand
 DNA Replication

5. DNA ligase:
ligase a linking enzyme that
catalyzes the formation of a covalent bond
joining fragments

Example: joining two Okazaki fragments together.

DNA ligase
Okazaki Fragment 1 Okazaki Fragment 2
5’ 3’

3’ Lagging Strand
5’
 3
DNA polymerase
• DNA 5 end
molecule 5

Daughter strand
polymerase Parental DNA synthesized
continuously
works in 5
3 Daughter
only one strand
synthesized
direction: 3
in pieces

P 5
5’ to 3’
• Telomere
sequences
are lost 5
P telomeres
with each 3

replication.
• Cancer, DNA ligase

aging Overall direction of replication

 Telomeres
Repeating, non-coding sequences at the end of
chromosomes = protective cap
 limit to ~50 cell divisions

growing telomerase
replication fork

Telomerase TTAAGGGTTAAGGGTTAAGGG

 enzyme extends telomeres

 can add DNA bases at 5 end
 different level of activity in different cells
• The information constituting an organism’s
genotype is carried in its sequence of bases

– The DNA is transcribed into RNA, which is

translated into the polypeptide

DNA

TRANSCRIPTION

RNA

TRANSLATION

Protein
 Mutations can change the meaning of
genes
• Mutations are changes in
the DNA base sequence
– caused by errors in DNA
replication or by mutagens
– change of a single DNA
nucleotide causes sickle-
cell disease