PROTEIN SYNTHESIS 1 DNA AND GENES DNA ■ DNA contains genes, sequences of nucleotide bases ■ Genes have different alleles. ■ These genes code for polypeptides (proteins) Genes & Proteins Proteins are made of amino acids linked together by covalent bonds called peptide bonds Dehydration synthesis or condensation reactions form the bonds 20 different amino acids exist 4 Amino Acid Structure 5 Polypeptides ■ Amino acid chains are called polypeptides 6 DNA Begins the Process ■ DNA is found inside the nucleus ■ Proteins, however, are made in the cytosol of cells by organelles called ribosomes ■ Ribosomes may be free in the cytosol or attached to the surface of rough ER 7 Starting with DNA ■ DNA ‘s code must be copied and taken to the cytosol ■ In the cytosol, this code must be read so amino acids can be assembled to make polypeptides (proteins) ■ This process is called PROTEIN SYNTHESIS 8 RNA Roles of RNA and DNA ■ DNA is the MASTER PLAN ■ RNA is the copy of the Master Plan 10 RNA Differs from DNA ■ RNA has a sugar ribose DNA has a sugar deoxyribose 11 Other Differences ■ RNA contains the base uracil (U) DNA has thymine (T) ■ RNA molecule is singlestranded DNA is double-stranded DNA 12 Structure of RNA 13 . Three Types of RNA ■ Messenger RNA (mRNA) copies DNA’s code & carries the 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 14 Messenger RNA ■ Long Straight chain of Nucleotides ■ Made in the Nucleus ■ Copies DNA & leaves through nuclear pores ■ Contains the Nitrogen Bases A, G, C, U ( no T ) 15 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 16 Ribosomal RNA (rRNA) ■ rRNA is a single strand 100 to 3000 nucleotides long ■ Globular in shape ■ Made inside the nucleus of a cell in the nucleolus ■ Associates with proteins to form ribosomes ■ Site of protein Synthesis 17 Transfer RNA (tRNA) ■ Clover-leaf shape ■ Single stranded molecule with attachment site at one end for an amino acid ■ Opposite end has three nucleotide bases called the anticodon 18 Transfer RNA amino acid attachment site U A C anticodon 19 Codons and Anticodons ■ The 3 bases of an anticodon are complementary to the 3 bases of a codon ■ Example: Codon ACU Anticodon UGA UGA ACU 20 The Genetic Code ■ A codon designates an amino acid ■ An amino acid may have more than one codon ■ There are 20 amino acids, but 64 possible codons ■ Some codons tell the ribosome to stop translating 21 The Genetic Code •Use the code by reading from the center to the outside •Example: AUG codes for Methionine 22 Name the Amino Acids ■ GGG? ■ UCA? ■ CAU? ■ GCA? ■ AAA? 23 TRANSCRIPTION AND TRANSLATION Pathway to Making a Protein DNA mRNA tRNA (ribosomes) Protein 25 Protein Synthesis The production or synthesis of polypeptide chains (proteins) Two phases: Transcription & Translation mRNA must be processed before it leaves the nucleus of eukaryotic cells 26 DNA RNA Protein Eukaryotic Cell Nuclear membrane DNA Transcription Pre-mRNA RNA Processing mRNA Ribosome Translation Protein 27 Transcription ■ The process of copying the sequence of one strand of DNA, the template strand ■ mRNA copies the template strand ■ Requires the enzyme RNA Polymerase 28 Question: What would be the complementary RNA strand for the following DNA sequence? DNA 5’-GCGTATG-3’ 29 Answer: ■DNA 5’-GCGTATG-3’ ■RNA 3’-CGCAUAC-5’ 30 Transcription ■ During transcription, RNA polymerase binds to DNA and separates the DNA strands ■ RNA Polymerase then uses one strand of DNA as a template to assemble nucleotides into RNA 31 Transcription ■ Promoters are regions on DNA that show where RNA Polymerase must bind to begin the Transcription of RNA ■ Called the TATA box ■ Specific base sequences act as signals to stop ■ Called the termination signal 32 RNA Polymerase 33 mRNA Processing ■ After the DNA is transcribed into RNA, editing must be done to the nucleotide chain to make the RNA functional ■ Introns, non-functional segments of DNA are snipped out of the chain 34 mRNA Editing ■ Exons, segments of RNA that code for proteins, are then rejoined by the enzyme ligase ■ A guanine triphosphate cap is added to the 5” end of the newly copied mRNA ■ A poly A tail is added to the 3’ end of the RNA ■ The newly processed mRNA can then leave the nucleus 35 Result of Transcription CAP New Transcript Tail 36 mRNA Transcript •mRNA leaves the nucleus through its pores and goes to the ribosomes 37 Translation ■ Translation is the process of of decoding the mRNA into a polypeptide chain ■ Ribosomes read mRNA three bases or 1 codon at a time and construct the proteins 38 Transcription Translation 39 Ribosomes ■ Made of a large and small subunit ■ Composed of rRNA (40%) and proteins (60%) ■ Have two sites for tRNA attachment --- P and A 40 Step 1- Initiation ■ mRNA transcript start codon AUG attaches to the small ribosomal subunit ■ Small subunit attaches to large ribosomal subunit mRNA transcript 41 Ribosomes Large subunit P Site A Site mRNA Small subunit A U G C U A C U U C G 42 Step 2 - Elongation ■ As ribosome moves, two tRNA with their amino acids move into site A and P of the ribosome ■ Peptide bonds join the amino acids 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 44 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 45 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 46 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 47 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 48 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 49 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 50 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 51 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 52 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 aa4 aa5 aa6 peptide bonds 53