PROTEIN SYNTHESIS: TRANSLATION AND THE GENETIC CODE
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1 PROTEIN SYNTHESIS: TRANSLATION AND THE GENETIC CODE HLeeYu Jsuico Junsay Department of Chemistry School of Science and Engineering Ateneo de Manila University 1
2 Nucleic Acids are important for their roles in the storage, transfer and expression of genecc informacon. HOW DO YOU TRANSFER INFORMATION TO THE NEXT GENERATION? HOW DO YOU DECODE THE INFORMATION AND MAINTAIN THE CELL S STRUCTURE AND FUNCTION? 2
3 Transfer and interpretacon of genecc informacon is described in the central dogma of molecular biology. Transla<on is required to convert the language of nucleic acids to the language of proteins. 3
4 TRANSLATION PROCESS 4
5 TranslaCon occurs in the ribosomes. Ribosomes are composed of two (2) major subunits. Each subunit is a complex structure of rrna and several proteins. 5
6 TranslaCon occurs in the ribosomes. Ribosomes are composed of two (2) major subunits. Each subunit is a complex structure of rrna and several proteins. 6
7 Ribosomes move along mrna templates deciphering the code, bring along adaptor molecules carrying amino acids capable of specific binding to mrna to decode informacon, and can catalyze the formacon of the pepcde bond. 7
8 Protein synthesis begins at the amino terminus. 8
9 Amino acids are carried by t RNA 9
10 Aminoacyl trna synthetase a[aches the amino acid to the t RNA (very specific) Mechanism and specificity Deacylase accvity "edits" and hydrolyzes misacylated aminoacyl trnas Despite common funccon, the synthetases are a diverse colleccon of enzymes Four different quaternary structures: α, α 2, α 4 and α 2 β 2 Subunits from 334 to more than 1000 residues 10
11 Amino acids are carried by trna. Each unique amino acid is carried by a specific trna Specificity is determined by a 3 nucleocde sequence in mrna called codons and the corresponding complementary sequence in trna called an<codons 11
12 This specific interaccon between the codon and a corresponding translacon to an amino acid is determined by the gene<c code. 3 nucleocdes (codon) encode an amino acid The code is nonoverlapping The code has no punctua6on The code is degenerate and universal 12
13 This specific interaccon between the codon and a corresponding translacon to an amino acid is determined by the gene<c code. GENOME ENGLISH analogy Nitrogenous Bases (ATCG) Letters (a,s,f,t,r,e, ) Codons Words Gene Sentences Chromosome Chapters Genome Book 13
14 14
15 Problem 1 What is the pepcde encoded by the following mrna? 3 AGAAUAUCGAAGCAGGGGUAGUGA 5 15
16 Problem 2 The following is the parent DNA strand. Assuming that splicing does not occur anymore ader transcripcon, give the pepcde it expresses. 5 -CTATAGAATCCCCCAATGACCACGCAT-3 16
17 TranslaCon is started by recognicon of start codon of the smaller ribosomal unit, start trna, inicacon factors (IF) and GTP. 17
18 NOTE: Prokaryote start is different from the eukaryote start. Prokaryote START fmet (formylmethionine) bound to inicator trna Recognizes AUG and somecmes GUG (but they also code for Met and Val respeccvely) AUG (or GUG) only part of the inicacon signal; preceded by a purine rich sequence Shine Dalgarno sequence 18
19 NOTE: Prokaryote start is different from the eukaryote start. 19
20 NOTE: Prokaryote start is different from the eukaryote start. Eukaryote START AUG nearest the 5 end is usually the start signal 20
21 When translacon is inicated, the large ribosomal subunit engages to complete the ribosomal catalycc sites 21
22 When translacon is inicated, the large ribosomal subunit engages to complete the ribosomal catalycc sites 22
23 When translacon is inicated, the large ribosomal subunit engages to complete the ribosomal catalycc sites 23
24 A second Aminoacyl trna molecule comes into the Asite and pep<dyl transferase creates the pepcde bond 24
25 A second Aminoacyl trna molecule comes into the Asite and pep<dyl transferase creates the pepcde bond 25
26 A second Aminoacyl trna molecule comes into the Asite and pep<dyl transferase creates the pepcde bond 26
27 A second Aminoacyl trna molecule comes into the Asite and pep<dyl transferase creates the pepcde bond 27
28 28
29 29
30 30
31 mrna is translated by lots of ribosomes, one ader the other. 31
32 Some ancbioccs inhibit translacon to kill off bacteria. Streptomyces venezuelae produces puromuycin 32
33 RECAP: 1. TranslaCon occurs at the ribosomes (large subunit and small subunit) 2. t RNAs carry specific amino acids to ribosomes. 3. Specific interaccons between mrna and trna allow for the decoding and translacon of nucleic acid to proteins 4. Ribosomes catalyze reaccon of pepcde bond formacon
34 POST TRANSLATIONAL MODIFICATIONS AND PROCESSING 34
35 Ader translacon, proteins fold due to intermolecular interaccons with water and itself. Chaperones facilitate and guide protein folding 35
36 Proteins are also biologically modified by a[aching different groups ProteolyCc cleavage (Zymogens) AddiCon of prosthecc groups (heme, etc.) PROTEIN Amino acid modificacon A[achment of Carbohydrates 36
37 Proteins are then transported to different parts of the cell by means of protein targekng. Protein that need to pass thru membranes have an extra amino acid sequence (cealled signal sequence) to tell the cell that they need to be transported. 37
38 Proteins also need to be concnuously degraded for funccon regulacon and removal of damaged or misfolded proteins. The protein s kiss of death marker is Ubiqui<n. 38
39 Proteins also need to be concnuously degraded for funccon regulacon and removal of damaged or misfolded proteins. The protein s kiss of death marker is Ubiqui<n. 39
40 REGULATION OF PROTEIN SYNTHESIS AND GENE EXPRESSION 40
41 Humans have about genes. Some are needed at all Cmes (cons<tu<ve genes). Some are needed only at specific points in a cell s life (inducible genes or repressible genes) 41
42 A gene cluster containing genes coding for different proteins of related funccons are called operons. 42
43 The RNA Polymerase DNA interaccon is Cghtly regulated by ac<vators and deac<vators. 43
44 The RNA Polymerase DNA interaccon is Cghtly regulated by ac<vators and deac<vators. 44
45 Regulatory proteins have discrete binding domains that allow them to recognize and bind to specific DNA. They parccipate via H bonding to DNA sequences. 45
46 About 80% of known regulatory proteins can be classified as : 1. Helix turn helix mo<f 46
47 About 80% of known regulatory proteins can be classified as : 1. Helix turn helix mocf; 2. Zinc finger mo<f 47
48 About 80% of known regulatory proteins can be classified as : 1. Helix turn helix mocf; 2. Zinc finger mo<f 48
49 About 80% of known regulatory proteins can be classified as : 1. Helix turn helix mocf; 2. Zinc finger mocf; 3. Leucine Zipper mo<f 49
50 About 80% of known regulatory proteins can be classified as : 1. Helix turn helix mocf; 2. Zinc finger mocf; 3. Leucine Zipper mo<f 50
51 Genes may also be silenced ader transcripcon (Post transcrip<onal gene silencing, PTGS) by use of RNAinduced silencing complexes (RISCs) or riboswitches. 51
52 Genes may also be silenced ader transcripcon (Post transcrip<onal gene silencing, PTGS) by use of RNAinduced silencing complexes (RISCs) or riboswitches. 52
53 Genes may also be silenced ader transcripcon (Post transcrip<onal gene silencing, PTGS) by use of RNAinduced silencing complexes (RISCs) or riboswitches. 53
54 Genes may also be silenced ader transcripcon (Post transcrip<onal gene silencing, PTGS) by use of RNAinduced silencing complexes (RISCs) or riboswitches. 54
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