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Lecture 12
Translation and
Proteins
1
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
2
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
3
Elongation of Polypeptide Chain
4
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
5
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
Alkaptonuria
Autosomal recessive
Darkening ears and nose
Benign arthritic conditions
6
Genes and Proteins
One gene: one enzyme
hypothesis
Proposed by George Beadle
and Edward Tatum (1940s)
Experiments in Neurospora
mutants
7
Amino Acids
Protein Structure
Tertiary Structure
Quaternary Structure
8
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
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
9
Protein Domains and Exon Shuffling
10