DNA VI - Translation and the Ribosome

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Karteikarten am DNA VI - Translation and the Ribosome, erstellt von J yadonknow am 27/12/2017.
J yadonknow
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J yadonknow
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Frage Antworten
Define a Ribosome - An organelle that provides the interface for translation of the genetic code into the formation of a functional protein
Describe the functions of a ribosome Provide an interface for mRNA and aminoacyl tRNAs Ribosomes catalyse the formation of peptide bonds
How does a ribosome facilitate tRNA binding? The ribosome has 3 tRNA binding sites: The (E) exit site The (P) peptidyl site The (A) aminoacyl site
What are ribosomes composed of? By mass ~35%/40% protein 60%/65% RNA
How is the size of a ribosome measured? Measured in Svedbergs (S) - Rate of sedimentation during centrifugation - Bigger the particle, faster it moves down a centrifugation tube
What are the subunits of a ribosome? Small 30s subunit, which binds to the mRNA being translated Large 50s subunit which catalyses peptide bond formation Size in total is 70s!!! Not 80s! (confusing, right?)
Draw a diagram of an E. Coli cell ribosome. 20nm high Large 50s subunit made of rRNA and proteins Small 30s subunit made of 16s rRNA and 21 proteins EPA sites
Human ribosome Large 60s subunit small 40s subunit EPA site overall size is 80s
How do mitochondrial ribosomes differ to human ribosomes Mitochondria have 55s ribosomes that are more similar to bacterial ribosomes than cytoplasmic ribosomes
Draw a diagram showing the process of translation Coding strand/promoter/coding region/terminator transcription (RNA polymerase) translation (tRNAs and ribosomes) Protein NH3+-COO-
Describe very briefly this translation mRNA is read 5'-3' Protein is synthesised NH3+-COO-
What is the first amino acid encoded in prokaryotes? N-formyl-methionine instead of methionine
Describe the process of initiation in E.Coli Small (30s) subunit assemble and binds to the ribosome binding site on the mRNA strand as a result of complementary base-pairing between it and the 16s rRNA.
What is this sequence known as? The Shine-Dalgarno sequence, exists just before the 1st AUG codon.
Describe the binding of the tRNA molecule to the ribosome F-met-tRNA^met binds to the initiation codon The 50s ribosome subunit then binds to generate the complete ribosome
Draw a diagram showing this binding corfu 06
Describe translation initiation in Eukaryotes First amino acid is methionine, not N-formyl-methionine 40s and met-tRNA^met combine prior to binding to mRNA
How does the ribosome bind to the mRNA molecule? There is no ribosome binding site on mRNA Instead, the 40s subunit recognises the cap at the 5' end of the mRNA
Draw a diagram you cunt well done you cunt
Draw a diagram translation elogation 1. Met-tRNA^met bound at P site 2. Binding of tRNA to A site, facilitated by EF-Tu/EF-Tα, requires 1 ATP equiv. 3. Peptidyl transferase reaction, peptide bond formed 4. Translocation, needs EF-G/EF-2 and ATP equiv. tRNA move along by 1 binding site 5.Release of 1st tRNA, binding of 3rd tRNA at A site, requires ATP equiv. 6. carries on until a termination codon is reached
What does the elongation cycle consist of? Repeated cycles of: Binding of aminoacyl-tRNA (needs GTPs) The peptidyl transferase reaction Translocation - movement of the ribosome one codon along the mNRA (needs GTP)
What is the result of each cycle? Incorporation of one amino acid into the protein
What terminates a cycle? Reaching a stop codon
Why does a stop codon terminate a cycle? As there is no tRNA molecule complementary to a stop codon
What is the energy expenditure during translation? Equivalent of 3 ATP used for each amino acid incorporated
What is each molecule equivalent of ATP used for? One to link the amino acid to its tRNA One is used when the aminoacyl-tRNA binds to the ribosome One is used to faciliate translocation
Peptidyl transferase diagram. Cunt. Well done. Prick.
What component of the ribosome catalyses the peptidyl-transferase reaction in E.Coli? The 23s rRNA
How do we know this? An E.Coli ribosome that has had ~95% of its proteins removed can still catalyse peptide formation X-ray crystallography shows that the active site for peptidyl transferase contains 23s rRNA but no proteins
What are elongation factors? Accessory proteins that are not part of the ribosome required for translation
Name the elongation factors required for prokaryotic elongation and its eukaryotic equivalent EF-Tu and EF-1α
Describe Step 1 of Elongation Deliver aminoacyl-tRNAs to the ribosome
Describe Step 2 of Elongation EF-G (EF-2 in eu) uses energy provided by the hydrolysis of GTP to move the ribosome along the mRNA during translocation Look at your notes here in detail to note the EPA site translocation
Compare eukaryotic and prokaryotic ribosomes Eu ribosomes larger Initiation is different: Recog of ribosomal binding sites (Pro) or cap (Eu) Elongation is very similar Both very sensitive to antibiotics and inhibitors
Name and describe the effects of a class of antibiotics that effect ribosomes Erythromycin/Clarithromycin target prokaryotic large ribosomal subunit Inhibits peptidyl transferase reaction & translocation
Name and describe the effects of a class of antibiotics that effect ribosomes cont. Cholramphenicol targets prokaryotic and mitochondrial large subunit Inhibits peptidyl transferase reaction
Name and describe the effects of a class of antibiotics that effect ribosomes (Cont. cont.) Ricin Targets Eu 60s subunit Catalytic action, removes a base fom rRNA, lethal dose in humans is approx 10^-4g
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