Fundamental Genetics

Lecture 12

Translation and
Proteins

John Donnie A. Ramos, Ph.D.

Department of Biological Sciences
College of Science
University of Santo Tomas

The Products of Transcription

‰ Messenger RNA (mRNA)
‰ primary structure
‰ linear sequence of RNA bases
‰ carries the genetic information in
the form of codons
Codons
‰ Ribosomal RNA (rRNA)
‰ assumes a 3D structure
(complexed with proteins)
‰ site of protein synthesis
‰ Transfer RNA (tRNA)
‰ assumes a cloverleaf structure
‰ carries amino acids from
cytoplasm to ribosomes

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 Ribosome
‰ Encoded by rDNA
(ribosomal gene)
‰ Synthesized by RNA
polymerase I in the
nucleolus
‰ Complex of RNA and
proteins (monosome)
‰ Prokaryotes: 10K/cell

Transfer RNA
‰ 75-90 nucleotides
‰ Nucleotides are post-transcriptionally
modified
‰ 2D cloverleaf structure (Rodbert Holley)
due to base pairing

‰ Amino acid binding site - ends

in CCA3’ and 5’G
‰ Anticodon loop – contains
RNA bases complementary to
3D Structure the codons
‰ Other loops serves as
recognition sites for enzymes
2D Structure during translation

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 Steps in Translation
1. Charging of tRNA
2. Initiation of translation
3. Elongation of polypeptide chain
4. Termination of translation
Charging of tRNA
‰ Loading of specific amino acid to its own tRNA
‰ Catalyzed by aminoacyl tRNA synthetase
‰ 32 different tRNA (despite the presence of 61
codons (bec. Of wobbling mechanisms)
‰ 20 different aminoacyl tRNA synthetases
‰ Isoaccepting tRNA – tRNA that binds to aa
‰ End product: aminoacyl-tRNA complex

Initiation of Translation

‰ Shine-Dalgarmo sequence (5’AGGAGG3’)

– sequence that precedes the first codon
in prokaryote m RNA
‰ Formylmethionine (fmet) – the first amino
acid of most polypeptides

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 Elongation of Polypeptide Chain

‰ Peptidyl site (P site) – contains the elongating

peptide
‰ Aminoacyl site (A site) – contains the amino acid to
be added
‰ Exit site (E site) – exit of uncharged tRNA
‰ Peptidyl transferase – catalyzes the formation of
peptide bond
‰ High efficiency (error rate 10-4)
‰ Rate of elongation: 15 aa/sec at 37°C (E. coli)

Protein Factors Involved in Translation

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 Termination of Translation
‰ Signaled by stop codons (UAG, UAA,
UGA)
‰ Release Factor 1 (RF1) – recognizes stop
codon UAA and UAG
‰ Release Factor 2 (RF2) – recognizes stop
codons UGA and UAA
‰ Release factors are GTP dependent
‰ Post-translational modification starts
after release from ribosome

Polyribosomes
‰ Single mRNA being used by different ribosomes for the process of
translation
‰ Also called polysomes
‰ A mechanism to produce more polypeptide (protein) copies

Translation of hemoglobin Translation in giant salivary

mRNA in rabbit reticulocyte gland cells of midgefly

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 Translation in Eukaryotes
‰ mRNAs stays in the cytoplasm for longer periods before degradation by
RNAses (hours)
‰ Ribosomes are much bigger
‰ mRNA is capped with 7-methyguanosine (7MG)
‰ mRNA contains an initiation sequence called Kozak sequence (ACCAUGG)
discovered by Marilyn Kozak
‰ Formylmethionine (fMet) is not required for initiation but met is often used
as a start codon
‰ More complex protein factors involved in different steps
‰ Elongating polypeptide enters the ER immediately as translation occurs

Proteins Form Phenotypes

Phenyketonuria
‰ Mental retardation
‰ Autosomal
recessive
‰ Inability of Phe to
converted to Tyr
‰ Accumulation of
Phe and its
derivatives in
cerobrospinal fluid

Alkaptonuria
‰ Autosomal recessive
‰ Darkening ears and nose
‰ Benign arthritic conditions

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 Genes and Proteins
‰ One gene: one enzyme
hypothesis
‰ Proposed by George Beadle
and Edward Tatum (1940s)
‰ Experiments in Neurospora
mutants

Genes and Proteins

‰ One gene: one protein (polypeptide chain)

‰ Not all protein are enzymes
‰ Example: Sickle Cell Anemia (mutant hemoglobin)

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 Amino Acids

Protein Structure

Primary Structure Secondary Structure

Tertiary Structure
Quaternary Structure

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 Post-translational Modifications
‰ Cleavage of formylmethionine
‰ Cleavage of signal peptides
‰ Acetylation of amino group
‰ Phosphorylation of certain amino acids
‰ Glycosylation
‰ Trimmining of polypetides
‰ Addition of metallic groups

‰ Molecular Chaperons – help proteins undergo correct protein folding

to become functional molecules.

Protein Function
‰ Structural Function ‰ Defense Function
‰ Collagen ‰ Antibodies
‰ Keratin ‰ Complement proteins
‰ Actin ‰ Catalytic Function
‰ Myosin ‰ Enzymes
‰ Regulatory Function ‰ Ribozymes
‰ Hormones ‰ Others
‰ Hemoglobin ‰ Histones
‰ Myoglobin ‰ Receptors

Enzyme Activity

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Protein Domains and Exon Shuffling

Structural domains of a fibronnectin molecule

DNA organization of an LDL receptor gene

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