Beruflich Dokumente
Kultur Dokumente
MOLECULAR BIOLOGY
• TYPES OF DNA
• TYPES OF RNA
• DNA REPLICATION
• RNA TRANSCRIPTION
• GENETIC CODE
• TRANSLATION.
DNA structure
Composition of DNA,
deoxyribonucleic acid, and
RNA, ribonucleic acid.
Slide 1
In slide 2, a nucleotide is seen to be
composed of 3 chemical groups: a
nitrogenous base or nitrogen containing
base, a sugar molecule & a phosphate (PO4) Composition of
group.
Slide 3
The H bonds are very systematic in how they form. Of
the 4 bases in DNA - adenine, guanine, cytosine &
thymine. Adenine always bind to Thymine & Guanine
always bonds with Cytosine.
Slide 4
Types of DNA
Minor groove wide and shallow narrow and deep narrow and deep
• In RNA, the sugar is ribose, which contains a hydroxyl group on the 2´-carbon.
•RNA chains are usually single-stranded and lack the continuous helical structure
of double-stranded DNA. However, RNA still has considerable secondary and
tertiary structure because base pairs can form in regions where the strand loops
back on itself.
•As in DNA, pairing between the bases is complementary and antiparallel. But in
RNA, adenine pairs with uracil rather than thymine. Basepairing in RNA can be
extensive, and the irregular looped structures generated are important for the
binding of molecules, such as enzymes, that interact with specific regions of the
RNA.
•The three major types of RNA (mRNA, rRNA, and tRNA) participate directly in
the process of protein synthesis. Other less abundant RNAs are involved in
replication
Messenger RNA (mRNA)
It comprises only about 5% of the RNA in the
cell yet is by far the most heterogenous type
of RNA in terms of size.
(1)In RNA, the sugar moiety to which the phosphates and purine and pyrimidine bases are
attached is ribose rather than the 2′-deoxyribose of DNA.
(2) The pyrimidine components of RNA differ from those of DNA. Although RNA contains the
ribonucleotides of adenine, guanine, and cytosine, it does not possess thymine. Instead of
thymine, RNA contains the ribonucleotide of uracil.
(3) RNA exists as a single strand, whereas DNA exists as a double-stranded helical molecule.
However, given the proper complementary base sequence with opposite polarity, the single
strand of RNA is capable of folding back on itself like a hairpin and thus acquiring
doublestranded characteristics.
(4) Since the RNA molecule is a single strand complementary to only one of the two strands of
a gene, its guanine content does not necessarily equal its cytosine content, nor does its
adenine content necessarily equal its uracil content.
(5) RNA can be hydrolyzed by alkali to 2′,3′ cyclic diesters of the mononucleotides, compounds
that cannot be formed from alkali-treated DNA because of the absence of a 2′-hydroxyl
group. The alkali lability of RNA is useful both diagnostically and analytically
Summary
DNA is
• a double stranded helix
• “rungs” joined together by H bonds
• phospodiester linkages between 5’ & 3’ ends of nucleotides; 5’ 3’ direction
• contains deoxyribose sugar
• contains A, T, C, G nucleotides
• found in the nucleus of cells; some is found in the mitochondria
RNA is –
• primarily a single stranded molecule
• linked by the same type of phosphodiester bonds that join DNA together; 5’ 3’
direction
• contains ribose sugar
• contains A, U, C, G nucleotides
• 3 types of RNA – messenger, ribosomal & transfer (mRNA, rRNA, tRNA)
• found in the cytoplasm; however it is manufactured in the nucleus, so some is found
there
DNA Replication
• The genetic information stored in the nucleotide sequence of DNA serves
two purposes.
• (1) It is the source of information for the synthesis of all protein molecules of
the cell and organism
• Both of these functions require that the DNA molecule serve as a template
—in the first case for the transcription of the information into RNA
• and in the second case for the replication of the information into daughter
DNA molecules.
• Replication of DNA occurs during the process of normal cell division cycles
• The mechanics of DNA replication was characterized in E. coli.
• Pol I is most abundant but has as its primary role to ensure the fidelity of
replication through the repair of damaged and mismatched DNA.
• Replication of the E. coli genome is the job of pol III which is much less
abundant than pol I, however, its activity is nearly 100 times that of pol I.
1. Primase
2. Processivity accessory proteins (dnaA binding protein)
3. Single strand binding proteins (SSB)
4. Helicase
5. DNA ligase
6. Topoisomerases
7. Uracil-DNA N-glycosylase
Steps in DNA Replication
• Steps involved in DNA replication. The figure above describes DNA
replication in an E coli cell, but the general steps are similar in
eukaryotes.
The sites of natural methylation (i.e. not chemically induced) of eukaryotic DNA
is always on cytosine residues that are present in CpG dinucleotides.
However, it should be noted that not all CpG dinucleotides are methylated at the
C residue.
This also helps prokaryotic cells to degrade invading viral DNAs. Since the viral
DNAs are not modified by methylation they are degraded by the host restriction
enzymes.
Chemotherapies Targeting Replication
Alkylating Agents
• The class of compounds that have been used the longest as anticancer
drugs are the alkylating agents which includes cyclophosphamide (Cytoxan,
Neosar), ifosphamide, decarbazine, chlorambucil (Leukeran) and
procarbazine (Matulane, Natulan).
• Some catalzye the cross-linking of bases in the DNA which prevents the
separation of the two strands during DNA replication.
• Alkylating agents act upon DNA at all stages of the cell cycle, thus they are
potent anticancer drugs. However, because of their potency, prolonged use
of alkylating agents can lead to secondary cancers, particularly leukemias.
Topoisomerases Inhibitors
Several classes of anticancer drugs function through interference with
the actions of the topoisomerases.
• The class of anticancer drugs that has been developed to interfere with
aspects of nucleotide metabolism is known as the antimetabolites.
• Those that inhibit DHFR are analogs of the vitamin folic acid and include
methotrexate (Trexall, Rheumatrex) and trimethoprim (Proloprim, Trimpex).
RNA transcription
Bacterial RNA transcription is described in
four steps:
• Note that the sequence in the RNA transcript and its polarity is the same as that in
the coding strand, except that the U of the transcript replaces the T of the gene.
Post transcriptional modification of RNA
• Poly-A tail is not transcribed from DNA, but rather added after transription
by poly-A polymerase
• The tail helps to stabilize the mRNA and facilitate its exit from the nucleus.
• When the poly-A mRNA enters the cytosol the tail is gradually shortened.
tRNA modification
• Addition of CCA
sequence to 3’-
terminal.
• This is catalyzed by
nucleotidyltransferase
Translation: Synthesis of Proteins
• After one ribosome binds and moves along the mRNA, translating the
polypeptide, another ribosome can bind and begin translation. The complex
of a single mRNA with multiple ribosomes is known as a polysome.
Several Antibiotic and Toxin inhibitors of Translation
Inhibitor Comments
Streptomycin inhibits prokaryotic peptide chain initiation, also induces mRNA misreading
Fusidic acid similar to erythromycin only by preventing EFG from dissociating from the
large subunit
Puromycin resembles an aminoacyl-tRNA, interferes with peptide transfer resulting in
premature termination in both prokaryotes and eukaryotes
Diptheria toxin catalyzes ADP-ribosylation of and inactivation of eEF-2, eEF-2 contains a
modified His residue known as dipthamide, it is this resudue that is the target
of diptheria toxin
Ricin found in castor beans, catalyzes cleavage of the eukaryotic large subunit
rRNA
Cycloheximide inhibits eukaryotic peptidyltransferase
Protein Synthesis
DNA passes on genetic
information so that
proteins can be
synthesized in the cell.
Proteins control many
different chemical
processes in the cell –
such as energy
production, intra &
extracellular
transportation, enzyme
synthesis, hormone
synthesis, and normal
cell maintenance.
Beside is the summary
of the process from DNA
replication to RNA
synthesis, also known
as transcription, and
translation, also known
as protein synthesis.
Note that once RNA is
made in the nucleus of
the cell, it leaves and
enters the cytoplasm,
where most cellular
proteins are made.