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1.
2.
3.
DNA polymerases
4.
5.
6.
Telomere replication
7.
Histone/chromatin assembly
2.
DNA template
3.
4.
Mg
2+
DNATemplate
EachstrandoftheparentDNAisusedasatemplate
template
tomakethenewdaughterstrand
DNAreplicationmakes2newcompletedouble
heliceseachwith1oldand1newstrand
ReplicationOrigin
Sitewherereplicationbegins
1inE.coli
1,000sinhuman
Strandsareseparatedtoallow
replicationmachinerycontact
withtheDNA
ManyATbasepairsbecause
easiertobreak2Hbondsthat3
Hbonds
Noteantiparallelchains
ReplicationFork
BidirectionalmovementoftheDNAreplicationmachinery
DNAPolymerase
Anenzymethatcatalyzes
theadditionofa
nucleotidetothegrowing
DNAchain
Nucleotideentersasa
nucleotidetriPO4
3OHofsugarattacks
firstphosphateoftriPO4
bondonthe5Cofthe
newnucleotide
releasingpyrophosphate
(PPi)+energy
DNAPolymerase
BidirectionalsynthesisoftheDNAdouble
helix
Correctsmistakenbasepairings
Requiresanestablishedpolymer(smallRNA
primer)beforeadditionofmorenucleotides
primer
Otherproteinsandenzymesnecessary
HowisDNASynthesized?
Originaltheory
Beginaddingnucleotidesatorigin
Addsubsequentbasesfollowingpairingrules
Expectbothstrandstobesynthesizedsimultaneously
ThisisNOThowitisaccomplished
Energy for this reaction is derived from the release of two of the
three phosphates of the dNTP.
2.
3.
Direction of synthesis is 5 to 3
DNA polymerase
Image credit:
Protein Data Bank
Polymerization (5-3)
Exonuclease (3-5)
Exonuclease (5-3)
#Copies
Yes
Yes
Yes
400
II
Yes
Yes
No
III
Yes
Yes
No
10-20
Eukaryotic enzymes:
Five common DNA polymerases from mammals.
1.
Polymerase
2.
Polymerase
3.
Polymerase
4.
Polymerase
5.
Polymerase
~245 bp in E. coli
Lagging strand
SSB Proteins
Okazaki Fragments
ATP
1
Polymerase III
2
3
Lagging strand
Helicase
+
Initiator Proteins
3
primase
Polymerase III
RNA Primer
3
base pairs
5
RNA primer replaced by polymerase I
& gap is sealed by ligase
Leading strand
Peter J. Russell, iGenetics: Copyright Pearson Education, Inc., publishing as Benjamin Cummings.
Peter J. Russell, iGenetics: Copyright Pearson Education, Inc., publishing as Benjamin Cummings.
2.
3.
Topoisomerases relieve
tensions in the supercoils,
allowing the DNA to continue
to separate.
Common in several
bacteriophages including .
2.
3.
4.
5.
2.
Cell will not enter the mitotic phase unless all the DNA has
replicated.
3.
4.
5.
Fig. 3.14
2.
3.
4.
Peter J. Russell, iGenetics: Copyright Pearson Education, Inc., publishing as Benjamin Cummings.
Fig. 3.17