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PROTEIN SYNTHESIS 1 Protein Synthesis The production (synthesis) of polypeptide chains (proteins) Two phases: Transcription & Translation 2 • SCI.9-12.B-4.3 - [Indicator] - Explain how DNA functions as the code of life and the blueprint for proteins. • SCI.9-12.B-4.4 - [Indicator] Summarize the basic processes involved in protein synthesis (including transcription and translation). 3 • Transcription – mRNA copying DNA, happens in the nucleus and mRNA must be edited or processed before it leaves the nucleus • Translation – when mRNA goes to ribosome and tRNA brings the correct amino acids to the ribosome 4 DNA RNA Protein Central Dogma DNA Eukaryotic Cell Transcription Nuclear membrane Pre-mRNA RNA Processing mRNA Ribosome Translation Protein 5 Pathway to Making a Protein DNA mRNA tRNA (ribosomes) Protein 6 Nucleic Acids 7 RNA 8 • SCI.9-12.B-4.1 - [Indicator] Compare DNA and RNA in terms of structure, nucleotides, and base pairs. 9 How DNA and RNA are alike: • Both made of monomers called nucleotides. • Both had A, G and C bases 10 *RNA Differs from DNA 1. RNA DNA 2. RNA DNA 3. RNA DNA has a sugar ribose has a sugar deoxyribose contains the base uracil (U) has thymine (T) molecule is single-stranded is double-stranded 11 . *Three Types of RNA • Messenger RNA (mRNA) carries genetic information to the ribosomes • Ribosomal RNA (rRNA), along with protein, makes up the ribosomes • Transfer RNA (tRNA) transfers amino acids to the ribosomes where proteins are synthesized 12 Making a Protein 13 *Genes & Proteins Proteins are made of amino acids linked together by peptide bonds 20 different amino acids exist Amino acids chains are called polypeptides Segment of DNA that codes for the amino acid sequence in a protein are called genes 14 Two Parts of Protein Synthesis Transcription makes an RNA molecule complementary to a portion of DNA Translation occurs when the sequence of bases of mRNA DIRECTS the sequence of amino acids in a polypeptide 15 • SCI.9-12.B-4.3 - [Indicator] - Explain how DNA functions as the code of life and the blueprint for proteins. • SCI.9-12.B-4.4 - [Indicator] Summarize the basic processes involved in protein synthesis (including transcription and translation). 16 *Genetic Code DNA contains a triplet code Every three bases on DNA stands for ONE amino acid Each three-letter unit on mRNA is called a codon Most amino acids have more than one codon! There are 20 amino acids with a possible 64 different triplets The code is nearly universal among living organisms 17 18 19 *What is the enzyme responsible for the production of the mRNA molecule? 20 *RNA Polymerase Enzyme found in the nucleus Separates the two DNA strands by breaking the hydrogen bonds between the bases Then moves along one of the DNA strands and links RNA nucleotides together 21 Question: What would be the complementary RNA strand for the following DNA sequence? DNA 5’-GCGTATG-3’ 22 Answer: • DNA 5’-GCGTATG-3’ • RNA 3’-CGCAUAC-5’ 23 *Processing Pre-mRNA • Also occurs in the nucleus • Pre-mRNA made up of segments called introns & exons • Exons code for proteins, while introns do NOT! • Introns spliced out by splicesomeenzyme and exons re-join • End product is a mature RNA molecule that leaves the nucleus to the cytoplasm 24 *Messenger RNA (mRNA) • Carries the information for a specific protein • Made up of 500 to 1000 nucleotides long • Sequence of 3 bases called codon • AUG – methionine or start codon • UAA, UAG, or UGA – stop codons 25 Messenger RNA (mRNA) start codon mRNA A U G G G C U C C A U C G G C G C A U A A codon 1 protein methionine codon 2 codon 3 glycine serine codon 4 isoleucine codon 5 codon 6 glycine alanine codon 7 stop codon Primary structure of a protein aa1 aa2 aa3 peptide bonds aa4 aa5 aa6 26 Transfer RNA (tRNA) amino acid attachment site methionine amino acid U A C anticodon 27 • SCI.9-12.B-4.3 - [Indicator] - Explain how DNA functions as the code of life and the blueprint for proteins. • SCI.9-12.B-4.4 - [Indicator] Summarize the basic processes involved in protein synthesis (including transcription and translation). 28 *Ribosomes • Made of a large and small subunit • Composed of rRNA (40%) and proteins (60%) • Have two sites for tRNA attachment --- P and A 29 Translation • Synthesis of proteins in the cytoplasm • Involves the following: 1. mRNA (codons) 2. tRNA (anticodons) 3. ribosomes 4. amino acids 30 *Translation • Three steps: 1. initiation: start codon (AUG) 2. elongation: amino acids linked 3. termination: stop codon (UAG, UAA, or UGA). Let’s Make a Protein ! 31 http://highered.mcgrawhill.com/olcweb/cgi/pluginpop.cgi?it=swf ::535::535::/sites/dl/free/0072437316/12 0077/micro06.swf::Protein%20Synthesi s 32 http://sciencenetlinks.com/interactives/pro tein.html 33 mRNA Codons Join the Ribosome Large subunit P Site A Site mRNA A U G Small subunit C U A C U U C G 34 Initiation aa1 aa2 2-tRNA 1-tRNA anticodon hydrogen bonds U A C A U G codon G A U C U A C U U C G A mRNA 35 Elongation peptide bond aa1 aa3 aa2 3-tRNA 1-tRNA anticodon hydrogen bonds U A C A U G codon 2-tRNA G A A G A U C U A C U U C G A mRNA 36 aa1 peptide bond aa3 aa2 1-tRNA 3-tRNA U A C (leaves) 2-tRNA A U G G A A G A U C U A C U U C G A mRNA Ribosomes move over one codon 37 aa1 peptide bonds aa4 aa2 aa3 4-tRNA 2-tRNA A U G 3-tRNA G C U G A U G A A C U A C U U C G A A C U mRNA 38 aa1 peptide bonds aa4 aa2 aa3 2-tRNA 4-tRNA G A U (leaves) 3-tRNA A U G G C U G A A C U A C U U C G A A C U mRNA Ribosomes move over one codon 39 aa1 peptide bonds aa5 aa2 aa3 aa4 5-tRNA U G A 3-tRNA 4-tRNA G A A G C U G C U A C U U C G A A C U mRNA 40 peptide bonds aa1 aa5 aa2 aa3 aa4 5-tRNA U G A 3-tRNA G A A 4-tRNA G C U G C U A C U U C G A A C U mRNA Ribosomes move over one codon 41 aa4 aa5 Termination aa199 aa3 primary structure aa2 of a protein aa200 aa1 200-tRNA A C U terminator or stop codon C A U G U U U A G mRNA 42 *End Product –The Protein! • The end products of protein synthesis is a primary structure of a protein • A sequence of amino acid bonded together by peptide bonds aa2 aa1 aa3 aa4 aa5 aa199 aa200 43 • SCI.9-12.B-4.3 - [Indicator] - Explain how DNA functions as the code of life and the blueprint for proteins. • SCI.9-12.B-4.4 - [Indicator] Summarize the basic processes involved in protein synthesis (including transcription and translation). 44 *Types of Mutations 1. Point mutation – one DNA bases changes. Sometimes this mistake is corrected by an enzyme. Types: a. insertion – where a base is added b. deletion – where a base is lost c. substitution – where a base is changed 45 Substitution Wild type A mRNA U G A 5 Protein A G U U G G C U A A 3 Lys Met U Phe Gly Stop Amino end Carboxyl end Base-pair substitution No effect on amino acid sequence U instead of C A U G A A G U Lys Met U G G Phe Missense A U U U Gly A A Stop A instead of G U G A A G U Lys Met U U A Phe G U U Ser A A Stop Nonsense U instead of A A U Met G U A G U U U G G C U A A Stop 46 • *The change of a single nucleotide in the DNA’s template strand – Leads to the production of an abnormal protein Wild-type hemoglobin DNA 3 Mutant hemoglobin DNA 5 C T T In the DNA, the mutant template strand has an A where the wild-type template has a T. G U A The mutant mRNA has a U instead of an A in one codon. 3 5 T C A mRNA mRNA G A A 5 3 5 3 Normal hemoglobin Sickle-cell hemoglobin Glu Val Figure 17.23 The mutant (sickle-cell) hemoglobin has a valine (Val) instead of a glutamic acid (Glu). 48 * 2. Frameshift mutation – insertion or deletion of a nucleotide (or base) changes the group of three bases that code for the amino acid ACT CAT TAG GAG (first T is deleted) ACC ATT AGG AG 49 Wild type mRNA 5 A A G U A Met Protein G U U Lys G U G C U Gly Phe A A 3 Stop Amino end Carboxyl end Base-pair insertion or deletion Frameshift causing immediate nonsense Extra U A G U A U Met A G U U U G C U A Stop Frameshift causing extensive missense Missing U A G G U A A Met G U Lys U G G C U A A Ala Leu Insertion or deletion of 3 nucleotides: no frameshift but extra or missing amino acid A A U Met A G Missing G U U Phe U G G Gly C U A A Stop 50