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The Genetic Code and Transcription: Flow of Genetic Information

The document discusses the flow of genetic information from DNA to protein. It covers the genetic code, including the discovery that it uses triplets of nucleotides, is degenerate, and universal across organisms. Key events included the discoveries by Nirenberg and others that codons specify amino acids. The document also discusses transcription, including the key parts of prokaryotic and eukaryotic transcription as well as post-transcriptional processing of eukaryotic pre-mRNA through capping, polyadenylation, splicing, and other mechanisms.

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Renz L. Salumbre
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681 views10 pages

The Genetic Code and Transcription: Flow of Genetic Information

The document discusses the flow of genetic information from DNA to protein. It covers the genetic code, including the discovery that it uses triplets of nucleotides, is degenerate, and universal across organisms. Key events included the discoveries by Nirenberg and others that codons specify amino acids. The document also discusses transcription, including the key parts of prokaryotic and eukaryotic transcription as well as post-transcriptional processing of eukaryotic pre-mRNA through capping, polyadenylation, splicing, and other mechanisms.

Uploaded by

Renz L. Salumbre
Copyright
© Attribution Non-Commercial (BY-NC)
We take content rights seriously. If you suspect this is your content, claim it here.
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Download as PDF, TXT or read online on Scribd
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Fundamental Genetics

Lecture 9

The Genetic Code


and Transcription

John Donnie A. Ramos, Ph.D.


Dept. of Biological Sciences
College of Science
University of Santo Tomas

Flow of Genetic Information

1
The Genetic Code
‰ Linear form (mRNA derived from DNA)
‰ Triplet codons (triplets of ribonucleotides coding for 1
amino acid)
‰ Unambiguous (1 codon = 1 amino acid only)
‰ Degenerate ( 1 amino acid can be specified by several
codons)
‰ Contains specific start and stop codons
‰ Commaless (no breaks once translation starts until the
stop codon is reached)
‰ Non-overlapping (single reading frame)
‰ Universal (same ribonucleotide used by all organisms)

The Discovery of the Genetic Code


‰ Francois Jacob and Jacques Monod
(1961) – messenger RNA (mRNA)
‰ Sydney Brenner (1960s) – codon in
triplets (minimal use of the 4 mRNA
bases to specificy 20 aa) (43=64)
‰ Francis Crick – frameshift mutations
alters the codons
‰ Mariane Manago and Severo Ochoa -
polynucleotide phosphorylase
(synthesis of RNA without template)-
paved the way to the production of
RNA polymeres in cell free-systems

2
The Discovery of the Genetic Code
‰ Marshall Nirenberg and J. Heinrich Matthaei (1661) – codons
‰ used cell-free protein synthesizing system and polynucleotide
phosphorylase
‰ RNA Homopolymers (UUUUUU…, AAAAAAA…, CCCCCC…, GGGGG…)
‰ UUU (Phenylalanine)
‰ AAA (Lysine)
‰ CCC (Proline)
‰ RNA Mixed Copolymers

1A:5C (1/6 A: 5/6C)

The Triplet Binding Assay


‰ Developed by M. Nirenberg and P. Leder (1964)
‰ Mimics the in vivo translation of proteins where a mRNA-tRNA-
ribosome complex is formed when all three macromolecules are
allowed to interact.

3
Repeating Copolymers
‰ Developed by Gobind Khorana (1960s)
‰ Synthetic long RNAs with repeating sequences

Results of Repeating Copolymers

4
The Universal Genetic Code

‰ Degeneracy
‰ Wobble
Hypothesis
‰ Start codon
(N-formylmethionine)

‰ Termination
codons
‰ Universal
Viruses
Bacteria
Archaea
Eukaryotes

Exceptions to the Universal Code

5
Transcription
‰ Uses DNA as a template
‰ Catalyzed by RNA polymerase
(holoenzyme of 500 kD)
‰ αββ’σ subunits
‰ Sense strand / template strand –
DNA strand used as a template
for transcription
‰ Promoter region – DNA sequence
recognized by σ factor to initiate
transcription (60 bases).
(upstream of a gene)
‰ TATA box (Pribnow box) –
TATAAT sequence
‰ Sigma factor (σ70, σ28, σ32, σ54)

Transcription
‰ RNA polymerase don’t need
primers
‰ Elongation in 5’ to 3’ direction
‰ Rate in E coli: 50 bases/sec at
37°C
‰ Termination is a function of rho
(ρ) factor – hexameric protein
interacting with the end of a
gene
‰ Polycistronic mRNA – bacterial
mRNA containing information for
the synthesis of proteins of
related function
‰ Monocystronic mRNA –
eukaryotic mRNA containing
information for a single protein.

6
Eukaryotic Transcription
‰ Features of eukaryotic transcription different from prokaryotic
transcription:
‰ Transcription inside the nucleus under the direction of 3 different
RNA polymerases

‰ Presence of protein factors (promoters, enhancers, etc.) binding to


the upstream portion of a gene (cis-acting elements) during
initiation step.
‰ Presence of post-transcriptional regulation.

Cis -acting Elements


‰ TATA Box (Goldberg-Hogness Box)
‰ Located 30 bases upstream from the start of
transcription (-30)
‰ Consensus sequence: TATAAAA
‰ Facilitates denaturation of helix because it is AT-
rich region

‰ CAAT Box
‰ Located 80 bases upstream from the start of
transcription (-80)
‰ Consensus sequence: GGCCAATCT
‰ Influence the efficiency of the promoter

7
Trans -acting Factors
‰ Transcription factors – facilitates template
binding during the initiation of transcription
‰ Example:
‰ TFIID (TATA-binding protein or TBP) – binds to
TATA-box

Post-transcriptional Processing
‰ 7-methylguanosine cap (7mG)
‰ Protection from nucleases
‰ Role mRNA transport across the
nuclear membrane
‰ 3’ cleavage site:
‰ AAUAAA
‰ Failure of 3’ cleavage results to
absence of poly A tail
‰ Split genes – contains intervening
sequences

8
RNA Splicing

‰ Ribozyme – RNA with


catalytic activity
‰ Self-excision process –
process of RNA splicing or
intron removal.
‰ Transesterification –
interaction between
guanosine and the
transcript.
‰ 2 successive
transesterification processes

The Spliceosome

‰ Alternative splicing
‰ Small nuclear ribonucleoproteins
(snRNP or snurps) – bonds to GU
or AG sites of introns
‰ 2 transesterification processes
‰ Snurps form a loop (lariat) in the
branch point region
‰ Produces isoforms of proteins

9
RNA Editing

‰ Substitution editing
‰ changes in the nucleotide bases of a given mRNA
‰ Common in mitochondrial RNA and chloroplast RNA
‰ Example: Apoliprotein B (Apo B) – C to U change CAA
to UAA
‰ Insertion / deletion editing
‰ addition or removal of nucleotide sequences
‰ Common in mitochondrial RNA or guide RNA (gRNA)

10

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