281 lec17 2019 - hageneticshagenetics.org/hh/wp-content/uploads/2019/09/281_lec17_2019.pdfthe trna...
TRANSCRIPT
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Lecture 17:
Translation: The tRNA and rRNA
Course 281
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AIMS
• Understand the structure and function of tRNA.
• Understand the process in which the tRNA carries an amino acid.
• Understand the components and structures of the ribosomes of prokaryotes and eukaryotes.
• Understand the process of making the ribosome from genes to their final structure.
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Translation – the process
What is translation?
Use the genetic code in the mRNA that reads 5’ ➔ 3’ to make a protein that reads N ➔ C.
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Translation – the process
What do we need to translate the genetic code?
1. mRNA 2. tRNA 3. Amino acids 4. Ribosomes
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Transfer RNA (tRNA)
• tRNA carries the amino acid to the ribosome to make protein.
• There are specific tRNA for each codon and amino acid.
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Transfer RNA (tRNA)
• The codon in the mRNA has a complementary sequence in the tRNA and it is called Anti-codon.
• Why is it called anti-codon?
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tRNA characteristics• It is 75-90 nucleotide in sequence.
• tRNA folds to form a specific shape called cloverleaf.
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tRNA characteristics
• How is the shape of tRNA formed?
• Contains three loops and the loop that contain the anticodon is called anticodon loop.
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tRNA characteristics
• There are two ends in the tRNA: • 5’ end • 3’ end
• The 3’ end is where the amino acid is attached and it is called the amino acid attachment site.
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tRNA characteristics
tRNA genes are found in multiple copies in the cell
(Why?)
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tRNA-amino acid
• Adding an amino acid to a tRNA is called aminoacylation or tRNA charging.
• An enzyme called aminoacyl-tRNA synthetase adds the correct amino acid to the corresponding tRNA.
• The process produces a charged tRNA or aminoacyl tRNA.
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tRNA-amino acidAminoacylation or tRNA charging
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tRNA-charging
• tRNA charging uses ATP as a source of energy.
• Each amino acid has a specific aminoacyltransferase.
• The 3’ nucleotides of tRNA is always CCA in all tRNAs.
• The amino acid binds to the 2’ or 3’ sugar of adenine (A) of the 3’ CCA.
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tRNA-charging
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Ribosomes
• Peptide synthesis and translation of the genetic code takes place on ribosomes.
• Ribosomes attach to mRNA and charged tRNA to make polypeptide chains.
• Both in prokaryotes and in eukaryotes the ribosome is made of:
Large subunit Small subunit
• Each subunit is composed of rRNA and ribosomal proteins.
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Bacterial ribosomes
• Bacterial ribosomes are called 70S ribosomes.
• The 70S ribosome is composed of two subunits:
• Large subunit (50S): • 23S rRNA • 5S rRNA
• Small subunit (30S): • 16S rRNA
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Bacterial ribosomes
• The S numbers do not add up correct?
• WHY? 50 S
30 S
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Eukaryotic ribosomes
• Eukaryotic ribosomes are called 80S ribosomes.
• The 80S ribosome is composed of two subunits:
• Large subunit (60S): • 28S rRNA • 5.8S rRNA • 5S rRNA
• Small subunit (40S): • 18S rRNA
80 S
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The rRNA contributes to the structure and function of the ribosomes.
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Bacterial ribosomal genes
• Both prokaryotic and eukaryotic rRNA is coded in the DNA by genes called ribosomal DNA (rDNA) or rRNA transcription units.
• In bacteria (E. coli), 7 rRNA transcription units are scattered through out the chromosome.
• Why many copies?
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Bacterial ribosomal genes
• Each rRNA transcription unit is composed of:
16S 23S 5S
• The transcription of these genes produces a precursor rRNA (pre-rRNA).
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Bacterial ribosomal genes
Spacers
• The pre-rRNA transcribed is 5’ 16S-23S-5S 3’ with non-rRNA sequences in between.
• The non-rRNA sequences are called spacers.
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Bacterial ribosomal genes
Spacers
• Ribonucleases remove the spacers and release three rRNA separate molecules.
• Is this like intron splicing?
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Prokaryotic ribosome
Ribosomal proteins with the three rRNA make the ribosome two subunits.
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Eukaryotic ribosomal genes
• Eukaryotic rRNA is coded by rDNA genes.
• The rDNA genes are composed of units containing:
18S-5.8s-28S
• This unit is repeated in eukaryotic genomes 100-1000 times.
• Why?
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Eukaryotic ribosomal genes
• The rDNA repeat units get transcribed by RNA Pol I producing a pre-rRNA with spacers.
• Pre-rRNA: 5’ 18S-5.8s-28S 3’
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Eukaryotic ribosomal genes
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Eukaryotic ribosomal genes
• Ribonucleases process the rRNA unit and remove spacers.
• 5S rRNA is located in other location than the repeat unit.
• What is the effect of the location 5S gene on its transcription?
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Eukaryotic ribosomal genes
• 5S rRNA is transcribed independently by RNA Pol III.
• The ribosomal proteins and the four rRNA molecules make the eukaryotic ribosome.
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To think about
• Where does the polymerase come from?
• The number of copies of rDNA genes in prokaryotic and eukaryotic genomes.
• The difference between intron splicing in eukaryotic genes and spacer removal in prokaryotic and eukaryotic pre-rRNA processing.
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Stuff to know
tRNAtRNA loops
Anticodon loop
tRNA charging
Aminoacylation CCA
Aminoacyl-tRNA synthetase
Cloverleaf shapeRibosomes
70S ribosome
80S ribosome
28S rRNA
23S rRNA
5S rRNA
16S rRNA
5.8S rRNA
Ribosomal large subunit
Ribosomal small subunit
rDNARibosomal DNA
rRNA transcription unit
Pre-rRNA
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Expectations
• You know the structure of tRNA and the process of charging the molecule with amino acids.
• You know the ribosomes of prokaryotes and eukaryotes:
• Components
• Genes
• Structure
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For a smile