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PURINES
Chemical structure of nitrogenous bases
PYRIMIDINES
SUBSTITUTIONS
OXYGEN,AMINO GROUPS,METHYL GROUPS
• PURINES
– ADENINE = 6-aminopurine
– GUANINE = 2-amino,6-oxypurine
• PYRIMIDINES
– CYTOSINE = 2-oxy,4-amino pyrimidine
– URACIL = 2,4-dioxypyrimidine
– THYMINE = 2,4-dioxy,5-methylpyrimidine
Nucleosides
Purines = N9-C1 glycosidic bond
Pyrimidines = N1-C1 glycosidic bond
A. Polynucleotides
B. Polyribonucleotides – polynucleotides containing
ribose as opposed to de-oxyribose
3. Naturally occuring are called RNA
4. Bases found in RNA’s (w/ a few exception) guanine,
cytosine, adenine, and uracil
C. Differences between the Primary Structure of DNA and
RNA
6. DNA has a 2’-deoxyribose as its sugar moiety rather
than ribose
7. DNA has one different base-thymine (5-methyl-uracil)
8. RNA has one different base-uracil
Ways of Presenting the structure of
a Polynucleotide (abbreviated form)
A. 5’ p-T-A-C-G-G-G-C-G-A-T-T-T-G-G-GOH³’
B. 5’ pTpApCpGpGpGpCpGpApTpTpTpGpGOH³
DNA – THE GENETIC MATERIAL
A. BASIC STRUCTURAL CHARACTERISTICS:
A double Helical Structure as proposed by Watson and
Crick (1953)
3. TWO ANTI – PARALLEL POLYDEOXY RIBONUCLEOTIDE
CHAINS – Wound around each other with the bases inside of the
helix sugar and PO4 – outside
4. STACKING OF THE HYDROHOBIC RINGS – Result when the
planes of the bases are perpendicular to the axis of the helix
5. HYDROGEN Bonding – between one bases on one stand pairing
w/ another base on the anti-parallel strand
Chargaff’s rule: content of A = content of T
content of G = content of C
4. The helix is 2 nm in diameter
= bases are separated by 3.4 A° along the helix
axis, and each base is rotated 36° in relation to
the previous base.
=the helical structure repeats at intervals of 34 A°
(every 10 base – pairs)
=In solution, the helical structure of DNA probably
does not repeat every 10-base pairs but nearer
to every 11 with the bases inclined 20° from the
horizontal
GENETIC INFORMATION IS STORED AS THE
SEQUENCE OF BASES ALONG THE CHAIN
Secondary structure of DNA: The
Double Helix
Right- and left-handed Helices
A, B, Z forms of DNA
Tertiary Structure of DNA
Supercoiling
• Supercoiling in prokaryotic DNA
– Positive supercoils
– Negative supercoils
• Supercoiling in Eukaryotic DNA
– Formation of chromatin
• electrostatic attraction between the negatively
charged phosphate groups on the DNA and the
positively charged groups on the protein
Supercoiling in prokaryotic DNA
Supercoiling in Eukaryotic DNA
Denaturation of DNA
• The double helix is disrupted during DNA
replication,transcription,repair and
recombination
• Therefore the forces that hold the two
strands together are adequate for
providing stability and yet weak enough to
allow easy strand separation
DISSOCIATION AND REASSOCIATION OF THE
DOUBLE HELICAL CHAINS OF DNA
mRNA
Cap Poly-A tail
NMP
ATP
ADP
NDP
Ribonucleotide thioredoxin (SH)2 NADP
Reductase
thioredoxin (S-S) NASPH+H
dNDP
ATP
ADP
dNTP
FORMATION OF TMP
-In the formation of dTMP from dUMP, FH4 is the methyl donor which
simultaneously is oxidized to FH2, this requiring reduction by dihy
drofolate reductase.
-This reaction is most sensitive to the inhibitory effect of aminopterin or
amethopterin (methotrexate).
-Aminopterin is an analogue of folic acid thus competitively inhibiting
dihydrofolate reductase, preventing the formation of FH4, the active
form of the enzyme.
Hence, this drug is used in cancer chemotherapy because of TMP
synthesis, and ultimately DNA replication.
-Trimethoprim (present in Bactrim, Cotrimoxazole) has similar
mechanism of action but acts more on bacterial cells, hence used
as antibiotics.
-Trimethoprim is generally used in combination with sulfonamides, a
para-aminobenzonic acid (PABA) analogue. PABA is a constituents
of folic acid, hence the presence of sulfonamides will prevent folic
acid synthesis in bacteria (but not in man, as folic acid is ingested
pre-formed in man as a vitamin), thus inhibiting TMP synthesis and
DNA replication of bacterial leading to cell death
FORMATION OF TMP
dUTP dCPM
Hydrolase Deaminase
Ppi NH3
dUMP
N5 N10 methylene
FH4
Dihydrofolate Thymidylate
Reductase synthetase
FH2 dMTP
SALVAGE PATHWAYS FOR PURINE AND
PYRIMIDINE BASES
-important in cells which cannot carry out the de
novo pathways e.g. due to lack of enzyme PRPP
amidotransferase (rate limiting step in purine
nucleotide biosynthesis) as in red blood cells.
-more economical in terms of high energy
phosphates expenditure compared to the de
novo pathway.
-nucleosides, which can enter cells (nucleotides
cannot) are either converted to a nucleotide thru
the action of a kinase or degraded to a base by
nucleoside phosphorylase
ENZYMES
1. Hypoxanthine – guanine phosphoribosyl
transferase (HGPRTase) catalyzes the one-
step formation of hypoxanthine or guanine to
IMP or GMP respectively
Guanine + PRPP GMP+PPi
Hypoxanthine + PRPP IMP + PPi
Complete deficiency of this enzyme produces
Lesch-Nyhan Syndrome characterized by:
1. Hyperuricemia and gout
2. Psychomotor abnormalities
3. Mental retardation
4. Self mutilation
2. Adenine phosphoribosyl transferase (APRTase)
catalyzes the conversion of adenine to AMP
Adenine + PRPP AMP+PPi
3. Pyrimidine phosphoribosyl transferase catalyzes
the conversion of orotate, uracil and thymine to
their corresponding nucleotides.
Orotate+PRPP OMP+PPi
Uracil + PRPP UMP+PPi
Thymine + PRPP TMP+PPi
FORMATION OF TMP
-In the formation of dTMP from dUMP, FH4
is the methyl donor which simulatneously
is oxidized to FH2, this requiring reduction by
dihy drofolate reductase.
-This reaction is most sensitive to the inhibitory
effect of aminopterin or amethopterin
(methotrexate)
FORMATION OR DEOXYPYRIMIDINE
NUCLEOTIDES FOR DNA SYNTHESIS
dCTP dTMP
DNA dTDP
dTTP
SALVAGE PATHWAYS
FREE BASE (PURINES, PYRIMIDINES)
RIBOSE-1-P
PRPP PI
PPi nucleosides
NMP ATP
ATP Kinase
NDP
ATP Kinase
NTP
Degradation of Pyrimidine Bases:
Deaminase reductase
Cytosine Uracil Dihydrouracil
H20 NH4+ NADPH NADP
H20
B-Ureidopropionate
B-alanine
CO2+NH3
B-aminoisobutyrate
B-ureidoisobutyrate
B-alanine
CO2+NH3
B-aminoisobutyrate
B-ureidoisobutyrate
• A. A form
• B. B form
• C. Z form
• D. D form
This DNA form is seen physiologic conditions
where cell is well hydrated:
• A. A form
• B. B form
• C. Z form
• D. D form
TRUE regarding salvage pathways of
nucleotide metabolism:
• A. They utilize free bases as substrates for
nucleotide biosynthesis
• B. They take place only in the mitochondria
• C. They derive the ring from amphibolic
intermediates
• D. They are more efficient than de novo pathway
TRUE regarding salvage pathways of
nucleotide metabolism:
• A. They utilize free bases as substrates for
nucleotide biosynthesis
• B. They take place only in the mitochondria
• C. They derive the ring from amphibolic
intermediates
• D. They are more efficient than de novo pathway
The enzyme that catalyzes the committed
step in purine de novo pathway:
• A. Xanthine oxidase
• B. PRPP synthase
• C. PRPP glutamyl amido transferase
• D. HGPRTase
The enzyme that catalyzes the committed
step in purine de novo pathway:
• A. Xanthine oxidase
• B. PRPP synthase
• C. PRPP glutamyl amido transferase
• D. HGPRTase
Xanthine Oxidase inhibitor
• A. Azaserine
• B. Sulfonamides
C. Trimethoprim
• D. allopurinol
Xanthine Oxidase inhibitor
• A. Azaserine
• B. Sulfonamides
C. Trimethoprim
• D. allopurinol
The first purine nucleotide synthesized
during the de novo pathway
• A. AMP
• B. GMP
• C. IMP
• D. ADP
The first purine nucleotide synthesized
during the de novo pathway
• A. AMP
• B. GMP
• C. IMP
• D. ADP
Which enzyme participates in both salvage
and De novo pathway?
• A.DNA renaturation
• B. DNA denaturation
• C. dsDNA
• D. RNA
Hyperchromicity effect is observed in:
• A.DNA renaturation
• B. DNA denaturation
• C. dsDNA
• D. RNA
If the adenine content of a double-helical
DNA is 20% of the total bases, the cytosine
content would be:
A. 20%
B. 30%
C. 40%
D. 50%
E. 60%
If the adenine content of a double-helical
DNA is 20% of the total bases, the cytosine
content would be:
• 20%
• 30%
• 40%
• 50%
E. 60%
END OF LECTURE