MedCom's Notes For

STORAGE & EXPRESSION OF GENETIC INFORMATION

DNA

3-5 phosphodiester bond

3 hydroxyl 5 hydroxyl
Free hydroxyl at 3 end & free phosphate at 5 end
Written in 5 3 direction
Hydrophilic inside,hydrophobic outside
Anticancer drugs act at narrow groove
G & C , A & T (covalent,hydrogen,hydrophobic interactions)
ssDNA high absorbance at 260nm

TYPES OF DNA:
Type

Handed

Residues/360 turn

Direction

Location

right

10

Base perpendicular to
helical axis

In chromosomes

right

11

20 degree away perpend

to helical axis

Dehydrated form
of B

left

12

Zigaz manner

In regions of
alternating base
pairs,poly GC

In prokaryotes non histones proteins are present.

DNA SYNTHESIS IN PROKARYOTES:
Semiconservative replication,1st studied in E.coli
A: separation of DNA in Two On AT rich site,in eukaryotes on diff sites
B: Formation of replication fork
A: Proteins for Separation
1.DNA protein AT rich site(ATP dependent)
2.DNA Helicase forcing strands apart(ATP dependent)
3. SSBP co-operatively bind,not an enzyme,shift equilibrium b/w ssDNA
dsDNA, protects DNA from nuclease,keeps separate strands

 B: Supercoils
Cord twisted in direction of tightening coils,it will wrap on itself producing +ve
S.C
Cord twisted in direction of loosing coils,it will wrap around itself in opposite
direction,-ve S.C
=> solution is Topoisomerases.
TOPOISOMERASE 1

TOPOISOMERASE 2

Nuclease & ligase activity

Transient breaks & release breaks

Not ATP dependent

ATP dependent

Relax ve coils in prokaryotes

& +ve,-ve coils in eukaryotes

In both prokaryotes eukaryotes because

its for interlocked chromosomes.

DNA gyrase types 2, unusual property of introducing ve S.C into relaxed DNA
using ATP,in bacteria & plants because ve neutralize +ve
Etopside (anticancer) Tropiso 2
DNA gyrase is target of auinoclones,ciprofoloxin(antimicrobial)

C: Direction of DNA replication

Read 3 5
Synthesize/written 5 3
Leading strand
Strand being copied in direction of advancing replication fork
Lagging strand
Copied direction away from replication fork(okazaki fragments)
D: RNA Primer:
Polymerase needs RNA PRIMERS (short double strand region having RNA
bases complementary to DNA, 3 end free hydroxyl serving to accept 1st
Deoxynucleotide by DNA polymerase)
RNA primer is formed by Primase,specific RNA polymerase.
Diff RNA primers in lagging but only 1 in leading.
Substrate for this is pyrophosphate,5 ribonucleoside triphosphates.
Primase + preparing complex of protein = Primosomes
Synthesis of primer is also 5 3

E: Chain Elongation

 DNA polymerase 3 for chain elongation of DNA chain highly processing

enzyme due to its beta subunit forming ring that encircles & move along template
Proofreading is by DNA polymerase 3,35 exonuclease activity
F: Excision of RNA primer & replacement by DNA
DNA polymerase 1,53 exonuclease activity,hydrolytically remove RNA
primer
5 3 can remove group of nucleotide or 1 nucleotide
G : DNA ligase
Phosphodiester linkage b/w 5 phosphate by poly 3 & hydroxyl by poly 1 on
chain (ATP dependent)
EUKARYOTIC DNA REPLICATION
Diff with prokaryotic
Many origins
RNA primers removed by RNAase H & FEN1 rather than DNA poly
A. cell cycle
A) Mitosis
B) G1 phase
C) S phase
D) G2 phase
B. EUKARYOTIC DNA POLYMERASES
POLYMERASES

FUNCTION

PROOFREADIN
G
no

Pol alpha

Contains primase ,initiates

DNA synthesis

Pol beta

repair

Pol gamma

Replicates mitochondrial
DNA

Yes

Pol delta

Elongates okazaki
fragments

Yes

Pol epsilon

Elongates leading strands

Yes

No

C: Telomeres
-> Complexes of non coding+proteins at ends of linear choromosomes
1.Maintains structure
2.prevent attacks
3.allow repair system to distinguish true end from break
-> Several 1000 tandem sequences of AG 3T2 complementary to Cs & As
-> GT rich is longer than CA
A) Telomeres shortens with successive cell division,in cancer cell,stem cells,germ
cells they dont because of ribonucleoprotein telomerase.
B) Telomerase:
1. A protein acts as reverse transcriptase & short piece of RNA acta s
template
2.extends 3 end of DNA
C) RNA primer is synthesized by primase
D) 3 end of RNA serve as acceptor of DNA poly
E) RNA primer is removed
D. REVERSE TRANSCRIPTASES
Involved in replication of retroviruses (HIV) (transposons)
E.INHIBITION OF DNA SYNTHESIS BY ANALOGS
2,3 dideoxyinosin (diadenosine) /deoxyribose arabinose prevent chain
elongation
Cytosine Arabenoside anticancer
Adenine Arabenoside antiviral
Zidovudine terminates chain elongation
ORGANIZATION OF EUKARYOTIC DNA
Histones basic amino acids +ve charges lysine+arginine 5 types
H1,H2A,H2B,H3,H4
Nucleosome: H2A,H2B,H3,H4
N terminals may be acetylated,phosphorylates,methylated
Linker DNA 50 base pair longH1
Prenatal histones are conserved
DNA REPAIR
U.V rays fuse pyrimidines
High ionizing radiations double strand breaks
A) Mismatch in E.COLI is corrected by MUT proteins
Identification:
GATC sequence once every 100 units is methylated in prenatal
strand
Repair:
Endonuclease mix & exonuclease remove

 LUNCH syndrome / HNPCC (hereditary non polyposis colonorectal cancer)

A. By U.V:
Fusion of dimers usually thymines prevents DNA poly,removed by
UVrABC proteins
1. Recognized
By UVrABC eonuclease/UV specific endonuclease
UV & cancer
Pyrimidine dimer in skin XP cant repair damage in proteins required for
correction of UV damage
B) Base excision repair
Cytosine(deamination) uracil (by nitrous acid)
Adenine---(D.A) hypoxanthine
Guanine(D.A) xanthine
C) Removal:
Specific glycolases
Specific AP endonucleases
B. Repair of double stand breaks
1.Non homologous end binding repair
Some DNA lost,prone
2.homologous recombination
Much less prone
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