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Elongation Enzyme moves 5’ 3’.
Rate is about 60 nucleotides per second.
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Comment Each gene can be read by sequential RNA Polymerases giving several copies of RNA. Result - several copies of the protein can be made.
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Termination DNA sequence that tells RNA Polymerase to stop. Ex: AATAAA
RNA Polymerase detaches from DNA after closing the helix.
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Final Product Pre-mRNA This is a “raw” RNA that will need processing.
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Modifications of RNA 1. 5’ Cap 2. Poly-A Tail 3. Splicing
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5' Cap Modified Guanine nucleotide added to the 5' end.
Protects mRNA from digestive enzymes. Recognition sign for ribosome attachment.
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Poly-A Tail 150-200 Adenine nucleotides added to the 3' tail
Protects mRNA from digestive enzymes. Aids in mRNA transport from nucleus.
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Comment The head and tail areas often contain “leaders” and “trailers”, areas of RNA that are not read. Similar to leaders or trailers on cassette tapes.
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RNA Splicing Removal of non-protein coding regions of RNA.
Coding regions are then spliced back together.
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Introns Intervening sequences. Removed from RNA.
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Exons Expressed sequences of RNA. Translated into AAs.
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Spliceosome Cut out Introns and join Exons together.
Made of snRNA and snRNP.
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snRNA Small Nuclear RNA. 150 nucleotides long.
Structural part of spliceosomes.
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snRNPs ("snurps") Small Nuclear Ribonucleoprotiens
Made of snRNA and proteins. Join with other proteins to form a spliceosome.
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Ribozymes RNA molecules that act as enzymes.
Are sometimes Intron RNA and cause splicing without a spliceosome.
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Introns - Function Left-over DNA (?) Way to lengthen genetic message.
Old virus inserts (?) Way to create new proteins.
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Final RNA Transcript
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Translation Process by which a cell interprets a genetic message and builds a polypeptide.
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Materials Required tRNA Ribosomes mRNA
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Transfer RNA = tRNA Made by transcription. About 80 nucleotides long.
Carries AA for polypeptide synthesis.
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Structure of tRNA Has double stranded regions and 3 loops.
AA attachment site at the 3' end. 1 loop serves as the Anticodon.
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Anticodon Region of tRNA that base pairs to mRNA codon.
Usually is a compliment to the mRNA bases, so reads the same as the DNA codon.
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Example DNA - GAC mRNA - CUG tRNA anticodon - GAC
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Comment "Wobble" effect allows for 45 types of tRNA instead of 61.
Reason - in the third position, U can pair with A or G. Inosine (I), a modified base in the third position can pair with U, C, or A.
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Importance Allows for fewer types of tRNA.
Allows some mistakes to code for the same AA which gives exactly the same polypeptide.
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Aminoacyl-tRNA Synthetases
Family of Enzymes. Add AAs to tRNAs. Active site fits 1AA and 1 type of tRNA. Uses a “secondary genetic” code to load the correct AA to each tRNA.
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Ribosomes Two subunits made in the nucleolus.
Made of rRNA (60%)and protein (40%). rRNA is the most abundant type of RNA in a cell.
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Large subunit Proteins rRNA
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Both sununits
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Large Subunit Has 3 sites for tRNA.
P site: Peptidyl-tRNA site - carries the growing polypeptide chain. A site: Aminoacyl-tRNA site -holds the tRNA carrying the next AA to be added. E site: Exit site
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Translation Steps 1. Initiation 2. Elongation 3. Termination
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Initiation Brings together: mRNA A tRNA carrying the 1st AA
2 subunits of the ribosome
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Initiation Steps: 1. Small subunit binds to the mRNA.
2. Initiator tRNA (Met, AUG) binds to mRNA. 3. Large subunit binds to mRNA. Initiator tRNA is in the P-site
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Initiation Requires other proteins called "Initiation Factors”.
GTP used as energy source.
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Elongation Steps: 1. Codon Recognition 2. Peptide Bond Formation
3. Translocation
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