Gene expression Chromatin : Interphase Non coding DNA
Non-histone proteins 30,000 to 100,000 bp form loops or - Telomere
DNA - Enzymes of replication,transcription domains which are anchored on - centromere Chromosomes - Topoisomerases RNA pol scaffoldings - Nucleus of eukaryotic - Small amount of RNA - Orders of structures Telomere - Mitochondria - RNA pol complex - 10nm -- form 30 nm region of repetitive DNA at the end of a - Chloroplasts of plants coding and non-coding segments chromosome - Prokaryotes have single chromosome Five types of histones: H1 protects the end of the chromosome and non-chromosomal DNA as Core histones: (H2A, H2B, H3 and H4)2 Nucleosome assembly from deterioration plasmids H1 least tightly bound Factors: are consumed during cell division and - DNA sequence replenished by an enzyme, the Plasmids Core histones - Acetylation/ telomerase reverse transcriptase DNA molecule is separate from the subject to modifications; which play role - Histone chaperone Nucleosome Prevents ends of chromosomes from chromosome in chromatin structure and function assembly protein 1 fusing can replicate independently - Acetylation(activation, inactivation of Assembly regulates the DNA replication, Genetic information gene transcription)(condensation of transcription, repair in humans, arrays of guanine-rich, six- transferable genetic elements, provide chromosome during replication) Super-packing of nucleosome: to eight-base-pair-long repeats TG rich mechanism for horizontal gene transfer - Methylation - In nuclei sequences; TTAGGG; - Antibiotic resistance - Phosphorylation - interaction of H1 with adjacent - toxins - ADP-ribosylation nucleosomes Eukaryotic telomeres normally terminate - As vectors in recombinant DNA - Mono-ubuquitylation (gene with 3 single-stranded-DNA overhang technology activation,repression, heterochromatic layout of the genome within the nucleus are disposable buffers blocking the ends gene silencing) is not random - specific regions of the chromosomes DNA - Sumoylation (transcription repression) Chromosomes demonstrate patterns Encodes the information that directs the (banding) Telomere shortening : been associated development of the organism Chromatin beads on a string with aging, malignant transformation Information are selectively expressed to Nucleosomes, the combination of DNA, Coding regions Telomeres form large loop structures be able to perform the functions histone, and other proteins that make Non-coding called telomere loops, or T-loops up chromosomes (10nm chromatin - Separate the functional domains(?) (single-stranded telomere DNA is held Gene: segment of DNA sequence fibril) to double-stranded DNA to form triple- corresponding to a single protein (single A 30 nm condensed chromatin fiber Barr body stranded called a displacement loop or catalytic or structural RNA molecule) consisting of nucleosome arrays in their X-inactivation D-loop) Gene : most compact form only in somatic cells disrupting the double-helical DNA and - by specifying the protein the cell is to one of the two X chromosomes base pairing manufacture ; 3 levels of chromatin organization - the DNA sequence dictates the entire 1. nucleosomes Karyogram of human male showing the Centromere: chemistry of the cell, including its the "beads on a string" structure classic metaphase chromatin structure site for attachment for proteins that link form and behavior ( therefore, the (chrmatin) the chromosome to the mitotic spindle cell is constructed and controlled by A 30 nm condensed chromatin : most Metaphase proteins) compact form Chromatids (duplicates) of chromosome DNA components Higher-level DNA packaging into the arranged in equatorial plane <100,000 proteins encoded by ~1% Smallest known genome : mycoplasma metaphase chromosome Telomeres exonic DNA genitalium 2. Chromatin : Centromeres - Most of DNA : non-protein coding Human : 3B proposed structures of the 30 nm heterochromatin Some DNA sequences regulate Largest : ameba dubia 670 B chromatin filament - where the DNA is more condensed, expression during development, Linker DNA (yellow) no transcriptional activity differentiation, adaptation to Nucleosome nucleosomal DNA (pink) - Some will remain condensed environment segment of DNA (140bp)wound around 3. Histones : throughout the cell cycle - Binding sites for regulators a histone protein core (oxtamer) high content of lysine and arginine Euchromatin - Code for regulatory RNA Solenoid structure (+) - where the "active" genes are - Introns (24%) separate functional chromatin is organized into chromosome Form ionic bonds with negatively - usually this region is much less domains charged DNA condensed - Repeated sequences with no defined H1 : aid packing nucleosome functions H2A, H2B, H3, H4 Eukaryotic DNA : sequence classes Chromosomal recombination Origin of replication Eukaryotes (5, a-e) Unique sequence Rearrange genetic material Prepriming complex - Pol a: replication nuclear DNA - Single copy genes that code for Exchanges which occur in meiosis - Unwinding - d : with a proteins - Maintain separation - B and e : repir of nuclear DNA Repetitive sequence Unequal exchange of information 1. DnaA - G : in mitochondria - Many copies in a cell; transcriptionqlly Deletion Bind to sp sequence inactive, structural role in Insertion, duplication 2. Single-stranded DNA-binding prtns DNA polymerase chromosome (highly repetitive) Unequal crossover (SSB) ; Read parental strand 3 to 5 direction ~30% (human) Keep separated can only synthesize a new DNA strand in - Moderately repetitive: interspersed in Separation of the 2 complementary strands Protect DNA from nucleases a 5' to 3' manner unique at specific locations called origins (of 3. DNA helicase dRNT are precursors mobile jump into various sites replication) Bind to SS near fork Phosphodiester bond with 3 OH May end up in disease - Origin replication element, Move forward to unwind the double ORComplex, strands (requires ATP) the process of replication goes Mitochondrial DNA - multiple sites differently for the two strands , double Double stranded circular - Short sequence of exclusively AT As strands separate helix Genetic code slightly different from base pairs (consensus sequence-) - Chromosome will rotate - Leading genomic DNA bidirectional - Cause positive supercoils) - Lagging Of maternal origin DNA topoisomerases: remove supercoils Establish evolutionary relationships Origin of replication DNA polymerases between species Recognized by prepriming complex Type I topoisomerase can not initiate synthesis of 1% of total DNA DnaA protein: bind to specific Nuclease and ligase activities complimentary strand on a single Code for nucleotides sequences rich in ATbp Cut, allows to rotate at phosphodiester stranded template - mt ribosomal and 22 transfer RNA - ATP bond (1 strand); relieve supercoils Require primer - 13 proteins in Respiratory chain - Cause to separate Negative supercoils: fewer turns of the - RNA, 10 NT helix than relaxed DNA - Which serves as the first acceptor Replication Interphase in cell cycle Positive : contain more turns of the of nucleotide by DNA polymerase each strand of the original double- DNA structure allows easy access of helix than relaxed DNA at 3 OH stranded DNA molecule serves as transcription and repair factors to the Can copy in 3 to 5 direction and template for the reproduction of the DNA Type II produce new strand in the 5 to 3 complementary strand Varies according to the access required Bind to DNA double helix dirction Occurs during S-phase (synthetic phase) At looser structure Make transient breaks in both strands Cell cycle - (euchromatin) Allow second stretch to pass thru break Primase (RNA polymerase) - G-S-G-M then reseals Join RNA nucleotides without requiring - Interphase: Types of Chromatin (prokaryotes and eukaryotes) a preexisting strand of nucleic acid G1, S, G2 Heterochromatin : inactive Synthesizes short RNA (10) - Mitosis - Densely packed (interphase) Prokaryotes complimentary to DNA and results in Prophase, metaphase, anaphase, - Constitutive: inactive Type II isomerase of E. coli hybrid duplex (U-A; G-C) telophase - Facultative: sometimes are actively DNA gyrase Use ribonucleosides TP - cytokinesis transcribed - Able to introduce negative supercoils Creates RNA primer - Becomes active during embryogenesis to resting (neutralize positive that will - DNA-RNA hybrid Prophase Euchromatin : active occur) primer initiates DNA polymerase III This is the phase where the - uncondensed - Requires ATP chromosomes of a cell are first visible Target of quinolones Hybrid complex under a microscope. Active site Template with RNA Primer Metaphase DNA : relatively more sensitive to Relieve negative supercoil Complimentary to the DNA template chromosomes will line up at the cell digestion by nuclease Many anti-cancer drugs function by - U pairs with A equator. - Hypersensitive site : slightly upstream inhibiting topoisomerases, which are Anaphase . to the active site (100-300 bp) especially active in rapidly growing when the sister chromatids separate - This provides clue about the presence tumor cells away from each other. and location of trsanscription element Telophase. DNA polymerase when two nuclear membranes are Unwinding of DNA at the origin, and Prokaryotes (3, pol I-III) pulling away from each other and are synthesis of new strands, forms a - Pol1 : repair being formed. replication fork - Pol III: replicative Primosome Formed by discontinuous copying of Eukaryotic telomeres normally terminate 3rd strand pairs with one main strand (Pre-priming complex proteins, primase) parental strand that runs 3 to 5 away with 3 single-stranded-DNA overhang foming D-loop Initiates formation of Okazaki fragments from the fork - Protective by sequestering the - Recognizes specific sequences that RNA primer is placed on the DNA strand Telomerase overhang terminal direct synthesis of RNA primer, moves 3' to the origin of replication ribonucleoprotein polymerase Function along the lagging strand in 5 to 3 Primosome moves opposite from DNA maintains telomere ends by addition of DNA pol polymerization direction (opposite from that of DNA synthesis the telomere repeat TTAGGG synthesis on lagging) Components: Helicase Processive unwinding Binds to SSDNA displacing Ssbinding DNA pol I - protein component with reverse proteins Recognizes nick between 3 end of DNA transcriptase activity Topoisomerases Relieve torsion. coils and the 5 end of RNA primer - RNA component that serves as a DNA synthesis Removes RNA (moving in 53 direction template for the telomere repeat DNA primase Initiates Synthesis of Replaces with DNA, synthesizing in 5 when it elongates telomeres RNA primase DNA pol III 3 direction TERT (reverse transcriptase) can add a Can only copy a DNA template in the 3 six-nucleotide repeating sequence, 5'- Single-strand Prevemnt premature Proofreads the new chain using its 35 to 5 TTAGGG to the 3' strand of binding proteins reannealing of exonuclease acitivity produce new strand in 5 to 3 chromosomes (in all vertebrates) (SSB) dsDNA Add dRNT to the 3 end of the RNA Activity of DNA pol I - These TTAGGG repeats DNA ligase Seals okazaki primer 53 can synthesize DNA in 35 telomerase can bind the first few fragments To the 3 end of the growing chain 35 exonuclease activity that nucleotides of the template to the last Synthesis is called continuous since it proofreads new DNA telomere sequence on the chromosome involves one nucleotide being placed 53 exonuclease that removes RNA add a new telomere repeat (5'-GGTTAG- E coli Mammalian Function
right after another in a series primer 3') sequence
DNA pol III proofreading activity I Gap filling, synth of Poofreads and removes mismatched and Finally, the DNA fragments on the end replication problem during lagging replaces (by 3 to 5 exonuclease activity lagging strand are hooked together by replication II DNA proofreading, done in reverse not the 5to 3; the enzyme DNA ligase + ATP In lagging strand repair degrades the chain only from the end) 5 from synthesized by DNA pol III and RNA primer attach ahead of initiation Synthesis continues until it is near the 3 OH made by pol I site DNA repair primer where it is blocked (lagging DNA polyerase (III) makes DNA strand Mitochondrial DNA strand) DNA polymerase until an RNA primer synthesis Recognizes the gap between the 3 end Prokaryotes RNA is excised and the gap is filled by of the synthesized DNA (by pol III) and - Pol I (involved in repair) DNA polymerase I III Leading strand the 5 end of the RNA primer ahead - pol II To replace RNA with DNA there must be synth exonuclease activity removes the primer - pol III (replicative) another DNA in frnt of the RNA primer Replaces with DNA (DNA pol I) Eukaryotes At the end where the last RNA primer is Histones - a, replication of nuclear DNA attached, DNA pol 1 can not convert the During replication chromatin is in Nucleoside analogs - B, in repair of nuclear DNA RNA to DNA relaxed form Cytosine arabinoside - g, in mitochondria RNA is destroyed by enzymes Nucleosomes are displaced Adenine arabinoside (antiviral) - d acts with a in replication Histones remain loosely associated with Zidovudine, acyclovir - E repair of nuclear DNA Lagging strand: one DNA strand At Terminal RNA cannot be replaced Synthesis : conservative leading strand template 53 exonuclease activity (by DNA pol with DNA template strand of the DNA double helix I) When RNA is excised Replication that is oriented in a 3' to 5' manner - Can remove 1 NT at a time from DNA The gap is not filled Occurs in both strands simultaneously read 3'-5' to produce a 5'-3' nascent that is properly base-paired, RNT or Section of telomere is lost at the 5 Leading strand: continuous strand dRNT 5 to 3 endexonuclease degrades ~130 Lagging: semidiscontinuous Copied towards the fork - Also removes groups of altered NT in 210 nucleotides so the 5 end is 5to 3direction 53 (1-10NT at a time) for rpair of shortened and terminal gap is enlarged lagging strand template damages Form 3 overhang Nuclear RNA the coding strand of the DNA double add a new telomere repeat (5'-GGTTAG- RNA primers removed helix that is oriented in a 5' to 3' manner Prokaryotes: 3') sequence DNA Polymerase reads 3'-5' on the DNA polymerase I "reads" the Telomere forms T loops and associates Mitochondria: original DNA to produce a 5'-3' nascent fragments, removes the RNA with double strand : D loop RNA remains as part of the closed strand circular DNA structure Bidirectional mRNA Difference from DNA replication Tanscription unit : DNA segment - Prokaryotes : one site of origin Cap: methylated guanine triphosphate Ribonucleotides are used sequence that will be copied (the - Eykaryotes : multiple attached to 5 OH end Uses U coding sequence) semiconservative - N7 in guanine is methylatd Primer is not used Primary transcript : product - 2 OH of 1st 2nd ribose may also be Only a semegnt of gene is copied Error rate : 10-9 to 10-10 methylated No proofreading during transcription In ds DNA with many genes, a strand Corrected during replication Poly A contains 200 adenine nucleotides will serve as the template for some Post-replication repair attached to OH at 3 end Prokaryotes (E, coli) genes and coding for others Mutation rRNA associate with proteins and form RNA pol: synthesizes all except RNA Changes in DNA ribosomes primer (replication) Prokaryotic transcription occurs in the Result in permanent change of base Multiunits: cytoplasm alongside translation sequence tRNA - Core enzyme : 4 subunits 2 alpha, 1 transcription and translation can occur Uncorrected errors during replication Modified nucleotides: pseudouridine, B, 1B simultaneously Damage caused by oxidative dihydrouridine, ribothymidine 53 RNA pol activity in eukaryotes transcription occurs in deamination, radiation, chemicals, Cloverleaf Lacks specificity, cannot nucleus while translation occurs in the results in cleavae of strands or removal CCA sequence at 3 end carries amino recognize promoter region cytoplasm of bases, chemical alteration acid - a 5th unit: sigma factor, required for initiation of synthesis (recognize Termination Types of mutations Transcription promoter regions) RNA pol can recognize signal on DNA: Point production of RNA copies of the DNA Recognize termination rho independent factor - Substitution of one base for another RNA polymerase - ( in E. coli, specific termination Rho factor Insertion Highly selective factor : rho factor) Rho (p) factor aids in termination in - addition of NT within DNA sequence - Signals (in nucleotide sequence of some Deletions DNA) initiation - Removal of NT - Instruct RNA polymerase where, how RNA pol binds to a promoter region in In a region where the transcript forms a often, and where to stop the gene hairpin loop that slows down and cause Rearrangement of genes - Biochemical differentiation of tissues Promoters : process to pause Recombination: is due to selectivity of the - Specificity of transcription of the At palindrome sequence rich in G and C - Between homlogous DNA segments transcription process gene at base of the hairpin turn Transposition Transcripts may undergo modifications Pribnow or TATA box; TATAAT, 10 base After the turn, Us occur which binds - Movement of DNA segment to a non- (terminal additions, base modifications, upstream the start point weakly with the DNA A homologous site trimming, internal segmant removal, Second sequence : TTGACA, 35 NT Separation of the RNA from DNA - Transposons (jumping genes) mobile splicing) upstream from start point template genetic elements that facilitate movement of genes Modulation of synthesis may result in RNA pol binds to promoter, then Rho-dependent - (in bacteria: resistqnce) altered rates of protein synthesis and unwinding of DNA occurs Rho factor which has RNA-dependent thus metabolic and phenotypic changes Copying starts ATPase activity RNA : working copies of DNA How organisms adapt to changes in RNA pol recognizes site of Termination environment mRNA produced: polycistronic transcript Major classes Template strand : DNA to be copied which is translated as it is transcribed rRNA: 80% of total DNA template is copied I 3 to 5; RNA (non-coding) - (produces several different proteins) mRNA: Total 2-5% chain grows in 5 to 3 Coding strand: same sequence as RNA - E. coli mRNA degraded in minutes tRNA: 60 sp, 15% transcript rRNA : produced as large transcript, Small nuclear RNA and microRNA Steps cleaved to form the ribosomal units - mRNA splicing and gene regulation Initiation RNA polymerase adds one RNA tRNA - <1% Elongation nucleotide at a time to a growing RNA - Produced from large transcripts that Termination strand are cleaved Small amount of T is present in tRNA Post-transcription modification from 5 end to 3 end - Cleavage enzymes RNase RNA molecules have extensive base - Into mature and active RNA Antiparallel to DNA template pairing which produces 2ry and 3ry - Errors may be cause of disease The resulting 5' 3' RNA strand is Rifampicin inhibits initiation of structures important for RNA function identical to the coding DNA strand rranscription Ribozymes , remove and splice thymines are replaced with uracils in the - Binds to B-subunit preventin themselves RNA formation of phsphodiester bond Act as ribonucleases, Rnase P cleaves Dactinomycin binds to DNA intrferes tRNA precursor with movement of RNA pol Synthesis in eukaryotes Splicing Termination: In cytoplasm the rsubunits bind with RNA pol in nucleus - Small nuclear RNA complexed with Eukaryotes mRNA to form 80S - Recognize particular tpes or genes proteins (snRNP) involved in Poorly understood - Ribozymes : autocatalytic RNAs in RNA pol I : synthesis of precursor of cleavage and splicing Signals downstream rRNA precursors large rRNA in nucleolus (28,18,5.8S) RNA endonuclease cleaves (15 bases3) - snRNA : in fomation of mRNA RNA pol II : precursor of mRNA Splice point consensus AAUAAA - Ribozymes translated to proteins, small nuclear Left flank of intron has sequence AG followed by addition of As at 3 end - RNA metab RNA and used by viruses to produce followed by invariant GU (polyadenylation) - Splicing viral RNA Right flank , invariant AG is followed by - Endoribonucleases hydrolyze RNA pol III: small RNAs, tRNA, 5s rRNA, GU Hairpin loop causes mechanical stress aminoacyl ester, peptidyl snRNAs which breaks the U-A bonds transferase Mitochondrial RNA pol : resembles Splicing mechanism Rho factor acts as an ATP-dependent bacteria polymerases Consensus sequences at the 2 splice unwinding enzyme, moving along the tRNA junctions newly forming RNA molecule towards its Produced by RNA pol III Synthesis of mRNA - 20-40 nt upstream fom 3 splice 3' end and releasing it from the DNA Primary transcript are cleaved at 5 and Basal promoter region site template 3 ends some may have introns Transcription factors : proteins that bind Spliceosome : complex that converts Undergoes extensive modification RNA polymerase II transcipt to mRNA Inhibitors of RNA pol II - Pseudouridine, rbothymidine, - Consists of: small ribonucleoprotein Alpha amanitia dihydrouridine Promoter: complex (snurp) - Toxin that binds and inhibits RNA pol - Methylation, reduction, deamination, TATA (Hogness) box: sequence - Pimary transcript II (stops mRNA synthesis) rearranged glycosidic bonds TATATAA ; 25 bp upstream (-25) - Small nuclear RNAs (5, - Toxin produced from Amanita - Addition CCA at 3end by CAAT box : 70 bp upstream U1,U2,U5,U4,U6)) phalloides Nucleotidyl transferase GC-rich regions (GC box) often - proteins - Form complex with polymerase and between-40 and -110 upstream inhibit mRNA synthesis Translation snurp messenger RNA (mRNA) :information transcription factors combine around to Positions the RNA sequence for splicing RNA editing in mRNA carrier coding for the synthesis of form preinitiation complex hnRNA heterologous nuclear RNA : Information is changed at the mRNA proteins Helicase: melts DNA unedited mRNA Ex: apoB gene - mRNA carrying a single protein snRNA with sn ribonucleoprotein form - In liver : is transcribed into mRNA sequence (common in eukaryotes) is Initiation base pairs with end of intron and that synthesizes 100kD : apoB100 monocistronic RNA polymerase binds to core facilitates splicing - In intestine - mRNA carrying multiple protein promoters in the presence transcription SLE: antibodies against snRNA - Cytidine deaminase convertsCAA sequences (common in prokaryotes) factors codon to UAA at one site; instead of is known as polycistronic Splicing errors coding for glutamine, becomes Enhancers - Ex B-thalassemia termination signal; result is apoB48 Genetic code : DNA sequences that function in the Composed of codons stimulation of transcription rate Summary rRNA 3nt-word (3 nucleotide bases/codon) Located thousands of bp upstream or Initiation Genes are in nucleoli 5 to 3 end douwnstream from the start site - finding of the start sequence Preribosomal (45s) 64 (43) Silencers : - Purine nucleotide Prokaryotes: 23s, 16s, 5s 61 code for 20 AA function in inhibition of transcription Promoter clearance Eukaryotes: 28s, 18s, 5.8s 3 : do not code for specific AA : RNA polymerase II - RNAP moves away from P after - 5s : synthesized by RNA pol III, nonsense codon Primary transcript: hnRNA chain is 10-20nt down to modified separately Termination signals: UAG, UGA, UAA (heterogenous nuclear RNA transcription unit most common start codon is AUG which Contains exons and introns Elongation rRNA is read as methionine - Addition to 3 RNA polymerase I stop codons have been given names: hnRNA Termination Genes in nucleolus UAG is amber, UGA is opal (sometimes Capped at its 5 end - Signalled by sequence in template 45s precursor modified by: also called umber), and UAA is ochre polyA tail (20 200 nucleotides, is strand of DNA - Methylation - 1 error in every 10100 million added to the 3 end - Cleavages bases Sequence AAUAAA in hnRNA serves as Elongation : RNA pol synthesize - Produce: 18s, 28s bonded to 5.8s signal for cleavage and addition of the transcript , adds NT from RNT 18s rRNA forms 40s polyA tail +supercoils are relaxed by gyrase and 60S is formed by 28S, 5.8 S, and 5S - ATP serves as precursor supercoils by topoisomerases Features of genetic code At least 1 tRNA per AA Example: Releasing factor 1 recognizes that a Degenerate(redundant, AA is encoded Some AA have more tRNA bond between the tRNA and methionine srop codon is in the A site by > 1 codon) is broken (releases free energy) Release factor falls into A, cause Each of the 20 AA has at least one Aminoacyl tTNA Is used to form the peptide bond release codon AA reacts with ATP to form enzyme large sub-unit then moves to three - Many AA are encoded by many complex: aminoacyl-adenosine MP + 2Pi bases (one codon) towards the 3' end of ribosome encounters a termination codon Aminoacyl-AMP ester with 2 or 3 the mRNA sequence hydroxyl of tRNA specific for the AA to The ribosome moves the alanine tRNA In general the 3rd nt in a codon is less form aminoacyl-tRNA and AMP tRNA for met is on the E and is to the P site. important than the 1st 2 in determining released from the ribosome (will The polypeptide is released the specific AA Protein synthesis beceom recharged with methionine) The ribosome has no new codons read. Degeneracy is in the 3rd nt 3 steps tRNA for proline is in the P site Wobble 1. Initiation (b) The A site for the next tRNA is free The protein will now be further modified 2. Elongation tRNA for Alanine has the anti-codon in either the endoplasmic reticulum, Unambiguous 3. termination CGA golgi or secreted in a vesicle. Given a specific codon, only 1 AA Is ribosome checks that this is correct indicated Initiation Termination Unambiguous but degenerate Assembly of initiation complex The bond between the tRNA and Proline Hydrolysis of bond between peptide Given specific codon ; only 1 specific AA 2ribosomal subunits is broken and tRNA in the P will be incorporated mRNA Free energy is released releases the protein from P site Given specific AA more than 1 codon AminoacyltRNA specified by the first A peptide bond is formed between Ribosomal subunits dissociate from the may be used codon Proline and Alanine. mRNA tRNA can use the anticodon to recognize GTP The peptide chain will be folding and The two sub units move and separate more than 1 codon Initiation factors shaping. Initiation Polysomes : Nonoverlapping assembly, met tRNA to form preinitiation Codon recognition Multiple ribosome on the same mRNA at NT is used only once complex Complimentary and antiparallel one time Reading of the code does not involve Binding of mRNA to form 43s ( initiation - mRNA is read 5 to 3 by anticodon in overlap of codons complex) 3 to 5 orientation Post translational processing Start : AUG (5 end) Binds to ribosome 60s forms 80s wobble hypothesis Phosphorylation - Determines the reading frame positions met tRNA at P site - Mechanism in which tRNA can Glycosylation Stop codon: UGA, UAG, UAA recognize more than one codon for a ADP-ribosylatin Nucleotide sequence that initiates specific AA Hyddroxylation Not punctuated, commaless Shine-Dalgarno sequence (E. coli) - (5') base of the anticodon pair with Addition of groups No pause between codons Locted upstream of Start (AUG); either as appropriate: it "wobbles" Sequence that is complimentary to Secretory proteins : synthesized in Universal Shine-D is near 3 end of 16Sr (in 30Sr) Pairing proceeds from the 5' end of ribosomes attached to RER All organisms use same codon for same mRNA 5end and 3 end of 16S r form the codon. AA complimentary base pairs thus Once the first two positions are paired, Exception: tRNA in mitochondria reads 4 positioning of mRNA on the 30S subunit exact base pairing of the third position codons differently from cytoplasm is less critical - AUA Met Codon AUG is recognized by special The third (5') base of the anticodon can - UGA Trp initiator tRNA (facilitated by IF) which typically pair with either member of the - As stop carries N-formylated met (prok) or purine or pyrimidine pair in the codon as AGA nonformylated in euk appropriate: it "wobbles AGG example, the double-ringed G can pair Requires only 22 tRNA to read the Elongation with either a single-ringed U or C genetic code Proper aminoacyl tRNA binds to A site This allows mRNA to be translated with In cytoplasm : 31 tRNA Peptide bond formation fewer than the 64 tRNAs that would be Translocation of peptidyl-tRNA to the P required without wobble. Specific tRNA for each AA site At least 20 enzymes which recognize Termination and attach AA to specific tRNA Termination codon appears in the A site Aminoacyl tRNA synthetase (UGA, UAG, UAA) To adenosyl terminal of tRNA Not recognized by tRNA