Translation - Prokaryotes
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1 1 Translation - Prokaryotes Shine-Dalgarno (SD) Sequence rrna 3 -GAUACCAUCCUCCUUA-5 mrna...ggagg..(5-7bp)...aug Influences: Secondary structure!! SD and AUG in unstructured region Start AUG 91% GUG 8 UUG 1 Surrounding of SD and AUG!!! Ribosomal protein S1: present only in Gram-negatives (not in Gram-positives): binds to AU-rich sequences found in many prokaryotic mrnas nucleotides upstream of start-codon Translational coupling AUG UAA UAG UGA AUG AUG
2 2 Translation - Eukaryotes Start Codon mrna 5 -CAP...AUG Influences: Surrounding of AUG!!! Kozak Consensus...CC A / G CCAUGG... mammalian... A / T A A / C A A / C AAUGTC T / C... Yeast. gccgcc(a/g)ccaugg.. Wikipedia
3 3 Translation elongation - Codon usage - Secondary structures - Codon structure translational frameshifting AAAAAAAAAUCA Lys Lys Lys Ser AAAAAAAAAUCA Lys Lys Lys Ile
4 4 Taken from: J.E. Krebs, E.S. Goldstein, S.T. Kilpatrick; Lewin s Genes XI ; Jones&Bartlett Learning
5 5 Taken from: J.E. Krebs, E.S. Goldstein, S.T. Kilpatrick; Lewin s Genes XI ; Jones&Bartlett Learning
6 6 Universal Triplet Code rare exemptions Other mitochondrial codes Other codes in cellular chromosomes Universal Codon code Mycoplasma ParameciumEuplotes Yeast Protozoa Mammals UGA Stop Tryptophan Stop Cysteine Tryptophan Tryptophan Tryptophan UAA/UAG Stop Stop Glutamine Stop Stop Stop Stop AUA Isoleucine Isoleucine Isoleucine Isoleucine Methionine Methionine Methionine CUA Leucine Leucine Leucine Leucine Threonine Leucine Leucine AGA/AGG Arginine Arginine Arginine Arginine Arginine Arginine Stop The universal genetic code is used in the chromosomes of most cells, chloroplasts, plant mitochondria, and their viruses and plasmids. A few organisms use slightly different codes in their chromosomes (in the nucleus). The examples of these other nuclear codes are from Mycoplasma (Bacteria) and two different ciii ated protozoa (Eukarya). All nonplant mitochondria use variations of the universal code, whereas plant mitochondria use the universal code. The examples here are only a few of the different types known. 1
7 7 Taken from: J.E. Krebs, E.S. Goldstein, S.T. Kilpatrick; Lewin s Genes XI ; Jones&Bartlett Learning
8 8
9 9
10 10 Regulation of Gene Expression Escherichia coli Prokaryotes Lactose Metabolism Absence of lactose Presence of lactose Only few molecules of ß-galactosidase per cell about 5000 molecules of ß-galactosidase per cell Not enzyme is inhibited, enzyme synthesis is affected Detailed biochemical and genetic analysis Jacob, Monod, Pardee Nobel prize
11 11 Taken from: J.E. Krebs, E.S. Goldstein, S.T. Kilpatrick; Lewin s Genes XI ; Jones&Bartlett Learning
12 12 lac-operon Ort I Ort O Taken from: J.E. Krebs, E.S. Goldstein, S.T. Kilpatrick; Lewin s Genes XI ; Jones&Bartlett Learning
13 13 Heterogenote analysis o+ z+ o- z- Cis-configuration inducible o+ z- o- z+ Trans-configuration constitutive
14 14 Heterogenote analysis i+ z+ i- z- Cis-configuration inducible i+ z- i- z+ Trans-configuration inducible
15 15 Model for behaviour of heterogenotes laco located adjacent to lacz, mutation in laco results in loss of regulatory function when connected to lacz, no complementation by wt-allele in trans laci located upstream of lacz, mutation in laci results in maintenance of regulatory function in both configurations to lacz complementation by wt-allele laco laci DNA locus, mobile factor binds there and represses synthesis encodes a mobile factor (= protein) which binds at laco
16 16 Taken from: J.E. Krebs, E.S. Goldstein, S.T. Kilpatrick; Lewin s Genes XI ; Jones&Bartlett Learning
17 17 λ Gene isolation lac operon Φ 80 Isolation of Lac Repressor laci q mutant Binding studies
18 18 Taken from: J.E. Krebs, E.S. Goldstein, S.T. Kilpatrick; Lewin s Genes XI ; Jones&Bartlett Learning
19 19 Inducer: ß-1,6- allolactose (by product of ß-galactosidase produced by transglucosylation) ß-Galactosidase Permease Transacetylase Taken from: J.E. Krebs, E.S. Goldstein, S.T. Kilpatrick; Lewin s Genes XI ; Jones&Bartlett Learning
20 20 Mutant O c Mutation in laco prevents binding of LacI Repressor protein to Operator Taken from: J.E. Krebs, E.S. Goldstein, S.T. Kilpatrick; Lewin s Genes XI ; Jones&Bartlett Learning
21 21 Mutant I - Mutation in laci no binding capacity of LacI repressor protein Taken from: J.E. Krebs, E.S. Goldstein, S.T. Kilpatrick; Lewin s Genes XI ; Jones&Bartlett Learning
22 22
23 23 Taken from: J.E. Krebs, E.S. Goldstein, S.T. Kilpatrick; Lewin s Genes XI ; Jones&Bartlett Learning
24 24 Taken from: J.E. Krebs, E.S. Goldstein, S.T. Kilpatrick; Lewin s Genes XI ; Jones&Bartlett Learning
25 25 Taken from: B. Lewin, Essential Genes, Pearson Ed. International
26 26 Taken from: J.E. Krebs, E.S. Goldstein, S.T. Kilpatrick; Lewin s Genes XI ; Jones&Bartlett Learning
27 27 Taken from: J.E. Krebs, E.S. Goldstein, S.T. Kilpatrick; Lewin s Genes XI ; Jones&Bartlett Learning
28 28 LacI repressor has general low affinity to DNA Unspecific weak binding LacI repressor has high affinity to specific operon Region on DNA Specific strong binding Taken from: J.E. Krebs, E.S. Goldstein, S.T. Kilpatrick; Lewin s Genes XI ; Jones&Bartlett Learning
29 29 Taken from: J.E. Krebs, E.S. Goldstein, S.T. Kilpatrick; Lewin s Genes XI ; Jones&Bartlett Learning
30 30 Taken from: J.E. Krebs, E.S. Goldstein, S.T. Kilpatrick; Lewin s Genes XI ; Jones&Bartlett Learning
31 31 Negative Regulation
32 32 Negative Regulation Taken from: J.E. Krebs, E.S. Goldstein, S.T. Kilpatrick; Lewin s Genes XI ; Jones&Bartlett Learning
33 33 Positive Regulation Taken from: J.E. Krebs, E.S. Goldstein, S.T. Kilpatrick; Lewin s Genes XI ; Jones&Bartlett Learning
34 34 Taken from: J.E. Krebs, E.S. Goldstein, S.T. Kilpatrick; Lewin s Genes XI ; Jones&Bartlett Learning
35 35 Influence of Glucose on expression of lac Operon Glucose controls import of lactose and of other alternative carbon sources
36 36 Carbon Catabolite Regulation Cyclic AMP acts as an inducer CAP (CRP) protein is a positive acting regulator protein Taken from: J.E. Krebs, E.S. Goldstein, S.T. Kilpatrick; Lewin s Genes XI ; Jones&Bartlett Learning
37 37 Taken from: J.E. Krebs, E.S. Goldstein, S.T. Kilpatrick; Lewin s Genes XI ; Jones&Bartlett Learning
38 38 Regulation at translation level Taken from: B. Lewin, Essential Genes, Pearson Ed. International
39 39 Taken from: J.E. Krebs, E.S. Goldstein, S.T. Kilpatrick; Lewin s Genes XI ; Jones&Bartlett Learning
40 40 Attenuation Taken from: J.E. Krebs, E.S. Goldstein, S.T. Kilpatrick; Lewin s Genes XI ; Jones&Bartlett Learning
41 41 Taken from: J.E. Krebs, E.S. Goldstein, S.T. Kilpatrick; Lewin s Genes XI ; Jones&Bartlett Learning
42 42 Taken from: J.E. Krebs, E.S. Goldstein, S.T. Kilpatrick; Lewin s Genes XI ; Jones&Bartlett Learning
43 43 Antisense RNA Taken from: B. Lewin, Essential Genes, Pearson Ed. International
44 44
45 45 Taken from: B. Lewin, Essential Genes, Pearson Ed. International
46 46 Taken from: B. Lewin, Essential Genes, Pearson Ed. International
47 47 Taken from: B. Lewin, Essential Genes, Pearson Ed. International
48 48
49 49 Taken from: B. Lewin, Essential Genes, Pearson Ed. International
50 50 Taken from: B. Lewin, Essential Genes, Pearson Ed. International
51 51 Taken from: B. Lewin, Essential Genes, Pearson Ed. International
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