Genetic code The language of nucleic acids: letters and words (nucleotides and codons) Initiation and termination codons of translation

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1 Molecular Biology: From DNA to Protein (cont d) Transcription Purpose and sub-cellular compartment. Steps of transcription and the bio-molecules involved. Terminology: promoter and terminator DNA sequences Genetic code The language of nucleic acids: letters and words (nucleotides and codons) Initiation and termination codons of translation Translation or protein synthesis Purpose and sub-cellular compartment and its three steps. The interpreter the language of nucleic acids into the language of proteins Mutation Definition, sources, impact the protein product Use the genetic code table to translate a sequence of codons into a sequence of amino acids and the reverse as well as the sequence of normal and mutated proteins

2 Transcription Producing copies of the genetic messages in the form of RNA

3 Transcription of a gene 1. DNA: double stranded 2. Enzyme: RNA Polymerase 3. Monomers: RNA nucleotides 4. Steps: Initiation- Starts at promoter sequence Elongation Termination- Stops at termination sequence Promoter DNA RNA polymerase 1 Initiation 2 Elongation DNA of gene 3 Termination Growing RNA Terminator DNA Figure 10.9B Completed RNA RNA polymerase

4 RNA polymerase unwinds the two DNA strands Uses one strand as a template strand Polymerizes RNA nucleotides against the template strand following the base pairing rules Single-stranded messenger RNA (mrna) peels away from the template strand The DNA strands rejoin RNA polymerase T C C A A T RNA nucleotides A U C C A T A G G T T A Figure 10.9A Direction of transcription Newly made RNA Template Strand of DNA

5 Template and coding strands of DNA - The two strands of DNA have complementary sequences - (template and coding strand) - RNA transcripts are single stranded and are made Only one DNA strand, the template strand is copied into RNA If given the sequence of one of the strands, predict the sequences of the other two using the base-pairing rule

6 In-Class Activity Write down the sequence of the mrna transcript of the following DNA sequence, given the information about the two strands DNA: 5'...CCATGGATCTTAGCA...3' (template strand) 3'...GGTACCTAGAATCGT...5 (sense Strand)

7 THE FLOW OF GENETIC INFORMATION FROM DNA TO RNA TO PROTEIN The information carried by sequence of DNA bases constitutes an organism s genotype The DNA genotype is expressed as proteins, which provide the molecular basis for the phenotype

8 Genetic information written in a code that is translated into amino acid sequences The words of the DNA language are triplets of bases (3 bases long) called codons Each codons in a gene specify one amino acid sequence of the polypeptide

9 Protein synthesis Translation is the RNA directed synthesis of a polypeptide Translation of the language of nucleic acids into the language of proteins (amino acids) One codon one amino acid

10 DNA molecule Gene 1 Gene 2 Gene 3 DNA strand- template A A A C C G G C A A A A Transcription U U U G G C C G U U U U RNA Codon Translation Polypeptide Figure 10.7 Amino acid

11 The genetic code is the Rosetta stone of life U UUU UUC UUA Second base U C A G Phe Leu UCU UCC UCA UCG Ser UAU UAC Tyr UAA Stop UAG Stop UGU UGC Cys UGA Stop UGG Trp U C A G Nearly all organisms use exactly the same genetic code Figure 10.8A First base C A G CUU CUC CUA CUG AUU AUC Leu Ile AUA AUG Met or start GUU GUC GUA GUG Val CCU CCC CCA CCG ACU ACC ACA ACC GCU GCC GCA GCG Pro Thr Ala CAU CAC CAA CAG AAU AAC AAA AAG GAU GAC GAA GAG His Gln Asn Lys Asp Glu CGU CGC CGA CGG AGU AGC AGA AGG GGU GGC GGA GGG Arg Ser Arg Gly U C A G U C A G U C A G

12 In-class activity/genetic code Use the genetic code table to answer the following: 1. How many codons are there for leu (leucine)? 2. How many codons are there for Met (Methionine)? 3. How many codons are there for Phe (phenylalanine)? Draw a conclusion about the number of codons for amino acids. 4. How many stop codons are there? Answer the following questions using this genetic code: 5 -AUGACCCCUUUGUUAUACUAA-3 5. How long is this message in nucleotides? 6. Is this the information present in DNA or in mrna? Explain your answer. 7. Write down the sequence of amino acids coded for by the above stretch of nucleotides. how long is this polypeptide?.

13 Translating the genetic code Strand to be transcribed (template) DNA T A C T T C A A A A T C A T G A A G T T T T A G Transcription m-rna 5 - A U G A A G U U U U A G - 3 Start condon Translation Stop condon Figure 10.8B Polypeptide Met Lys Phe

14 - The coding sequence of mrna is between the start codon, which marks the translation initiation and a stop codon, which marks the end of translation Start of genetic message RNA nucleotides that are not part of the coding sequence End RNA nucleotides that are not part of the coding sequence Figure 10.13A

15 Translation Takes place in cytoplasm Involves: Ribosomes: Two subunits (each made of many proteins & r-rna) mrna t-rna (interpreter) 20 amino acids

16 Transfer RNA molecules serve as interpreters during translation Amino acid attachment site Hydrogen bond RNA polynucleotide chain Figure 10.11A Anticodon

17 Each trna molecule is a folded molecule bearing a base triplet called an anticodon on one end A specific amino acid is attached to the other end Amino acid attachment site Anticodon Figure 10.11B

18 A ribosome attaches to the mrna translates its message into a specific polypeptide aided by transfer RNAs (trnas) Protein Synthesis animation 5/animations.html# Translation Intro (Flash Animation)

19 Steps of protein Synthesis 1. Initiation a. Binding of mrna to small ribosomal subunit b. Binding of Met-tRNA to the initiation codon AUG on mrna c. Binding of large ribosomal subunit 2. Elongation 3. Termination

20 mrna, a specific trna, and the ribosome subunits assemble during initiation Initiator trna Met U A C A U G P site Start codon mrna Small ribosomal 1 subunit 2 Met U A C A U G Large ribosomal subunit A site Figure 10.13B

21 Steps of protein Synthesis 1. Initiation a. binding of mrna to small ribosomal subunit b. binding of Met-tRNA to AUG on mrna c. Binding of large ribosomal subunit 2. Elongation a. binding of a second trna to next codon b. formation of peptide bond c. sliding of ribosome by one codon Amino acids are added to the growing polypeptide chain until a stop codon is reached. 3. Termination Disassembly of the protein synthesis machinery

22 Elongation steps Polypeptide mrna P site Codons A site Amino acid Anticodon 1 Codon recognition mrna movement Stop codon 2 Peptide bond formation New Peptide bond Figure Translocation

23 Review: The flow of genetic information in the cell is DNA RNA protein The sequence of codons in DNA, via the sequence of codons in mrna spells out the primary structure of a polypeptide

24 Mutations are changes in the DNA base sequence Caused by errors in DNA replication or recombination, or by mutagens Normal hemoglobin DNA Mutant hemoglobin DNA C T T C A T mrna mrna G A A G U A Figure 10.16A Normal hemoglobin Glu Sickle-cell hemoglobin Val

25 Substituting, inserting, or deleting nucleotides alters a gene with varying effects on the organism. Normal gene mrna Protein A U G A A G U U U G G C G C A Met Lys Phe Gly Ala Base substitution A U G A A G U U U A G C G C A Met Lys Phe Ser Ala Base deletion U Missing A U G A A G U U G G C G C A U Figure 10.16B Met Lys Leu Ala His

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