Chapter 6 Expression of Biological Information (Part IV) By : Biology Unit Kedah Matriculation College Concept review… Expression of Biological Information DNA & genetic information DNA structure DNA as a carrier DNA replication Protein synthesis Gene regulation & expression concept Components Gene concept Mechanism Transcription Definition Models of DNA replication DNA replication process Enzyme involved Translation Learning outcome Explain translation in protein synthesis Describe the stages involved: i. initiation ii. elongation (codon recognition, peptide bond formation and translocation) iii. termination Translation Translation Definition Translation - a process - whereby the genetic information in mRNA strand - is translated into sequence of amino acids to form polypeptide/protein Occurs in cytoplasm Translation RNA direct the synthesis of polypeptide. cell interprets a genetic message and builds a protein accordingly. The message is a series of codons along an mRNA molecule and interpreter is transfer RNA Structures that involve in translation Ribosome tRNA triplet bases on tRNA: anticodon mRNA Structures that involve in translation: 1. tRNA tRNA transfer amino acid from cytoplasm to ribosome tRNA can be used repeatedly. Each type of tRNA molecule associates a particular mRNA codon with a particular amino acid. Shape: 2dimensional – cloverleaf shape. tRNA includes the anticodon, the specialized base triplet that binds to a specific mRNA codon. e.g.: mRNA codon UUU has AAA as its anticodon. From the other end of the Lshaped tRNA molecule protrudes its 3’end; the attachment site for amino acid. tRNA with amino acid; aminoacyl tRNA. 2. Aminoacyl-tRNA synthetase Catalyze the matching up of amino acid with the correct tRNA and using energy of ATP (activation of amino acids) Catalyzes the covalent attachment of amino acid to the 3’- OH of tRNA. Resulting aminoacyl tRNA Activation of aminoacyl tRNA 3. Ribosomes Facilitate the specific coupling of tRNA anticodons with mRNA codons during protein synthesis. A large and small subunit join to form a functional ribosome only when they attach to mRNA molecule. Each ribosomal subunit is consist proteins and ribosomal RNA (rRNA). Ribosome has TWO binding site for mRNA: P site (peptidyl-tRNA site) Holds the tRNA carrying the growing polypeptide chain. A site (aminoacyl-tRNA site) Holds the tRNA carrying the next amino acid to be added to the chain. Translation process Overall Process… 3 steps of translation process, synthesis of a polypeptide chain : 1. 2. 3. Initiation Elongation Termination 1. Initiation Bring together mRNA and tRNA bearing the first amino acid of polypeptide and two subunits of ribosome. Small ribosomal subunit binds to mRNA that being read from 5’ to 3’. Initiation Then initiator tRNA base-pairs with the start codon Arrival of a large subunit completes the initiation complex Initiation initiation factor: required to bring all the translation components together Initiator tRNA site in the P site of ribosome and vacant A site is ready for next tRNA. initiation complex of translation Ribosome (large) tRNA mRNA Ribosome (small) 2. Elongation Amino acid are added one by one to the initial amino acid. Each addition, occurs in 3 steps cycle. i. ii. iii. Codon recognition Formation od peptide bond Translocation i. Codon recognition mRNA codon in A site forms hydrogen bonds with the anticodon of an incoming molecule of tRNA carrying its appropriate amino acid. Elongation factors lead the tRNA into A site. hydrogen bonds ii. Peptide bond formation Polypeptide detaches from tRNA (in P site) to which it was bond and is link to amino acid carried by the tRNA (in A site) large ribosomal subunit catalyzes the formation of peptide bond between polypeptide extending from P site and newly arrived amino acid in A site. 2. Peptide bond Formation 3. Translocation iii. Translocation tRNA in P site dissociates from ribosome. tRNA in A site now attached to the growing polypeptide, is translocated to the P site iii. Translocation as the tRNA changes sites, its anticodon remains hydrogen bonded to the mRNA codon, allowing the mRNA and tRNA molecules to move as a unit. This movement, in turn, brings the next codon to be translated into A site. Elongation is repeated as each amino acid is added to the chain until the polypeptide is completed. Peptide bond formation 3. Termination Elongation continues until A site reaches termination codon. do not code for amino acids and stop translation. - No aminoacyl tRNA recognize Termination Protein called ‘release factor’ binds to the termination codon in A site. - freeing the polypeptide from the ribosome. - mRNA detaches - ribosome separates Initiation Large Unit Met Initiation codon AUG P A UAC 5’ UUG UGG UAC AUG GAA GAG Small Unit Initiation Complex Small Unit CAU … UGA 3’ ACU CUU Met UUU UAC 5’ UUG UGG UAC AUG GAA GAG Elongation CUC CAA ….UGA 3’ ACU GUG 5’ UUG UGG UAC Termination AUG GAA GAG STOP codon UAA UAG UGA CAU UGA 3’ 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 copyright cmassengale 41 mRNA U C C A U C A U G G G C U C C A U C G G A Codon 1 methionine Polyribosomes Polyribosomes/polysome A group of several ribosomes attached to, and translating the same messenger RNA simultaneously Increase the rate of same protein synthesis/protein produced rapidly more of same protein can be made simultaneously Animation RNA polymerase DNA Can you tell the story? amino acids exon intron tRNA pre-mRNA 5' GTP cap mature mRNA large ribosomal subunit 5' small ribosomal subunit poly-A tail aminoacyl tRNA synthetase 3' polypeptide tRNA E P A ribosome