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BIO20 Lecture Handouts On Molecular Genetics
BIO20 Lecture Handouts On Molecular Genetics
INFORMATION PATHWAYS
Test made in mouse DNA:
Gene is the smallest unit of inheritance. It is a 10% of the DNA consists of short lengths of less
segment of DNA that encodes the information than 10 base pairs that are repeated millions of
required to produce a functional biological product. times per cell. highly repetitive
20% of the DNA consists of about hundreds of base
The units of information are copied or converted pairs repeated at least a thousand times per cell.
into functional products through three major moderately repetitive
processes: The Central Dogma of Molecular 70% of the DNA are repeated only few times.
Genetics (unique sequence of base pairs)
Replication – copying of parental DNA to form Satellite DNA – most highly repeated sequence.
daughter DNA molecules having identical
nucleotide sequences. Junk DNA – some repetitive DNA.
Transcription – process by which parts of the coded Centromere – attachment site for proteins that
genetic message in DNA are copied precisely in the links chromosomes.
form of RNA.
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BIO20 Introduction to Biomimetic Engineering and Component Design
Central Dogma of Molecular Genetics
Replication is rapid.
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BIO20 Introduction to Biomimetic Engineering and Component Design
Central Dogma of Molecular Genetics
PROOF-READING IN REPLICATION
DNA replication is, by far, the most accurate of
known enzyme-catalyzed processes. The error rate
-9
per base pair per round of replication is about 10
-10
to 10 . The accuracy cannot be explained,
however, by the different binding energies between
Multiple proteins are required for DNA replication at correct and incorrect base pairs (this amounts to
a replication fork such as DNA helicases, DNA only a 100 to 1000 fold difference between correct
polymerases, topoisomerases, and DNA ligase. and incorrect pairing).
Some of these are multisubunit protein complexes.
Two cellular systems aid the fidelity of replication.
There are three major steps in DNA Replication: These include the following:
INITIATION 1. The 3'-5' exonuclease activity of DNA
polymerases acts as a "proofreading" system.
Helicases actively denature DNA and actively Incorrectly incorporated nucleotides are not readily
unwind duplex DNA strands. It provides this extended by the polymerase very efficiently,
function by catalyzing the ATP-dependent allowing them to be melted away to the 3'-5'
unwinding of double-strand DNA. exonuclease site of the polymerase for removal.
Topoisomerases are enzymes with a "swivel"
mechanism that can relieve torsional stress. 2. The mismatch repair system makes an additional
Bidirectional replication of the circular E. coli contribution to accuracy of about 100-fold. It works
chromosome unwinds about 100,000 base pairs per by scanning the newly replicated DNA, excising any
minute. At 10 bp per turn, this is 10,000 turns of residues that are not properly base-paired and
DNA per minute or over 160 per second. If there replacing them with the correct nucleotides.
were nothing to relieve this stress, DNA would be a
tangled mess in seconds. Relief of torsional stress
is essential for DNA replication to occur.
ELONGATION
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BIO20 Introduction to Biomimetic Engineering and Component Design
Central Dogma of Molecular Genetics
TERMINATION
DNA ligase - Covalently closes nicks in double- TRANSCRIPTION
stranded DNA. The nick must contain 3' hydroxyl
and 5' phosphoryl termini and the nucleotides being - process of RNA Synthesis
linked must be adjacent in a duplex structure and - copying the 5’ – 3’ DNA in the form 5’ – 3’ RNA
properly base-paired. DNA ligase functions to seal
Okazaki fragments in the lagging strand of DNA -synthesis of 5’ to 3’ RNA antiparallel to DNA
replication. template strand (thru base pairing):
Structure of RNA
tRNA
structure
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BIO20 Introduction to Biomimetic Engineering and Component Design
Central Dogma of Molecular Genetics
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BIO20 Introduction to Biomimetic Engineering and Component Design
Central Dogma of Molecular Genetics
Post-transcriptional Modification (Eukaryotes) c. RNA Splicing - After being capped, the pre-
A. Ribosomal RNA mRNA becomes complexed with a number of
- intermediate-sized pieces of rRNAs are small nuclear ribonucleoprotein particles
trimmed to produce the required rRNA (snRNPs), which are themselves complexes of
species. small nuclear RNAs (snRNAs) and special
splicing enzymes. The snRNP--pre--mRNA
complex is called a spliceosome. snRNAs
recognize and bind intron--exon splice sites by
Ribosomal RNA gene means of complementary sequences. Excision
of a single intron involves assembling and
RNA pol I disassembling a spliceosome. The sequence of
reactions can be summarized as follows:
45S
1. It begins with the attachment of the U1
(28S) (5.8S) (18S) snRNP to the G site at the 5' end of the intron.
2. The U2 snRNP then attaches at the branch
cleavage by RNases site.
3. Assembly of the spliceosome continues,
including the addition of several more snRNPs,
(28S) (5.8S) (18S) 4. The lariat loop in the intron is formed and the
Ribosomal RNAs two exons are joined. Splicing has now been
accomplished, and the products--a ligated
B. Transfer RNA mRNA and a looped intron--are released. As
-modifications include addition of –CCA the spliceosome disintegrates, the looped intron
sequence by nucleotidyltransferase on the 3’- is degraded, and the mRNA is exported from
terminal end of tRNAs the nucleus.
C. Messenger RNA
a. 5’ Capping - the first modification occurs at the
5' end of the pre-mRNA. A GTP residue is
added in reverse orientation and forms,
together with the first two nucleotides of the
chain, a structure known as a cap. The cap is
"decorated" by the addition of methyl groups to
the N-7 position of the guanine and to one or
two sugar hydroxyl groups of the cap
nucleotides. The cap structure serves to
position the mRNA on the ribosome for
translation.
b. Addition of poly-A tail – adenine nucleotide is
added after transcription by poly-A polymerase.
This modification helps stabilize the mRNA and
facilitate their exit from the nucleus.
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BIO20 Introduction to Biomimetic Engineering and Component Design
Central Dogma of Molecular Genetics
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BIO20 Introduction to Biomimetic Engineering and Component Design
Central Dogma of Molecular Genetics
Termination of Translation
THE STANDARD GENETIC CODE
Marshall Nirenberg and Philip Leder in 1964
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BIO20 Introduction to Biomimetic Engineering and Component Design
Central Dogma of Molecular Genetics
Consequence of Deletion:
Change in structure of nucleotide sequence of the FRAMESHIFT MUTATION – a shift in the
DNA reading frame, which produces a non-functional
protein
a. Mutation can be spontaneous or induced.
ATT GGC AAA GCA TCA AAC TTA GGG CCC
b. Mutation according to size:
Point mutation deletion of G at
Gross Chromosomal mutation codon 4
Effects: ATT GGC AAA CAT CAA ACT TAG GGC CC…
Lethal --- reading frame was changed
Branching in the evolutionary tree or specie
diversity C. Insertion – addition of one or more nucleotides
in the DNA
Point Mutation
Consequence of Deletion:
A. Substitution mutation FRAMESHIFT MUTATION – a shift in the
- single-base mutations, substitution of one or reading frame, which produces a non-functional
more nucleotides by the same number of protein
different nucleotides.
Deletion and Insertion Mutations are
Types: Transition and Transversion usually caused by Replication Slippage
Transition
purine to purine D. Exon Skipping – results from mutation at the
A G spliced site
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BIO20 Introduction to Biomimetic Engineering and Component Design
Central Dogma of Molecular Genetics
Deaminating agents
- removes amino groups from a base
Example: Sodium nitrite (used as preservative,
color enhancer and color fixative bacon, smoked
fish, tocino, etc.)
When ingested, NaNO2 is converted to nitrous acid
in acidic conditions. HNO2 removes functional
groups from Adenine, Guanine and Cytosine. Like
deamination of adenine, will result in the formation
of hypoxanthine, a base analogue of guanine.
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