05.

Genetic code and Translation

mRNA export to the cytoplasm

Reticulocyte lysates . Prepared from red blood cells (no nucleus) . Large quantities of Ribosomes . tRNAs . rNTPs . ATP . mRNA haemoglobin

O Cdigo Gentico

- - - - - - -

cada 3 nucleotideos = 1 codo Conjunto de codes = Quadro de leitura (Open Reading Frame ORF) 64 codes (4 x 4 x 4) = 61 para aminocidos 3 codes STOP Cdigo gentico e Universal Codes so seguidos sem interupo degenerado = + do que 1 codo para a maior parte dos aminocidos Codes especfico de iniciao = AUG Codes especficos de terminao = UAG (Amber); UAA (Ocare); UGA (Opal)

8 grupos de codes com os primeiros dois nucletidos idnticos e o terceiro pode ser qualquer um 7 pares de codes em que os dois primeiros so iguais e o terceiro e uma pirimidina (C,T,U) 5 pares de codes em que os dois primeiros so iguais e o terceiro e uma purina (A,G) 2 codes que so especficos 3 codes em que no tem amino cido associado

O Codigo gentico

Quadro de leitura (Open reading frame ORF)

5' 3' ! ATGCCCAAGCTGAATAGCGTAGAGGGGTTTTCATCATTTGAGGACGATGTATAA! TACGGGTTCGACTTATCGCATCTCCCCAAAAGTAGTAAACTCCTGCTACATATT! 3 ! ! ! ! ! !5!
5 1 ATG CCC AAG CTG AAT AGC GTA GAG GGG TTT TCA TCA TTT GAG GAC GAT GTA TAA !

M 2 3

*!

TGC CCA AGC TGA ATA GCG TAG AGG GGT TTT CAT CAT TTG AGG ACG ATG TAT ! C P S * I A * R G F H H L R T M Y! GCC CAA GCT GAA TAG CGT AGA GGG GTT TTC ATC ATT TGA GGA CGA TGT ATA ! A Q A E * R R G V F I I * G R C I !

Quadro de leitura (Open reading frame ORF)

Alteraes no cdigo gentico na mitocndria

Mitorcndria - Fosforilao oxidativa - Genoma circular 16,000 pb - Materno

Alteraes no cdigo gentico

- - - - UGA STOP=Triptofano Metionina Interna = AUG e AUA Metionina iniciadora = AUG;AUA;AUU;AUC Codes STOP = UAA;UAG;AGA;AGG

Traduo: overview

t-RNAs

t-RNA synthetases

Humanos 500 tRNA genes S 48 codes representados

Transfer RNA (tRNA) is the 'adaptor' molecule that enables the Genetic Code contained
in the nucleotide sequence of a messenger RNA (mRNA) molecule to be translated into
the amino acid sequence of a polypeptide chain. The key to this process lies in the specic
recognition of the correct tRNA molecule by an aminoacyl-tRNA synthetase enzyme which
attaches the correct amino acid for the tRNA to the acceptor stem at the 3' end of the molecule.

Existem so 48 tRNAs embora o cdigo gentico tem 64 codes. (61 funcionais) Emparelhamento wobble permite a terceira base do anticodo na posio 5 um emparelhamento imperfeito

Wobble pairing 3 end mRNA

Bases modificadas presentes em tRNAs

t-RNA synthetases

Aminoacyl-tRNA synthetases from all organisms belong to one of two classes depending on the amino
acid they are responsible for. Class I enzymes are generally (though not always) monomeric, and attach
the carboxyl of their target amino acid to the 2' OH of adenosine 76 in the tRNA molecule. Class II
enzymes are generally dimeric or tetrameric, and attach their amino acid to the 3' OH of their tRNA,
except for phenylalaninyl-tRNA synthetase which uses the 2'

t-RNA synthetases

Most cells make twenty different aminoacyl-tRNA synthetases, one for each type of amino acid. These
twenty enzymes are widely different, each optimized for function with its own particular amino acid and
the set of tRNA molecules appropriate to that amino acid. The one shown here, which charges aspartic
acid onto the proper tRNA (entry 1asz), is a dimer of two identical subunits (colored blue and green, the
two tRNA molecules are colored red). Others are small monomers or large monomers, or dimers, or even
tetramers of one or more different types of subunits.

Aminoacyl-tRNA-synthetases

Ativao do aa

Transferncia do aa para o tRNA

Aminoacil-tRNA sintetasas

Controlo de qualidade

Iniciao da traduo requer um tRNA especfico

Ribosome structure

Os rRNAs determinam a estrutura dos ribossomas

Prokaryotic translation Initiation So Formil-Met tRAN pode entrar no Site E

Iniciao

Eucaryotic translation initiation

Elongao

Atividade peptidil-transferase

Transferncia do aa para o prximo tRNA no enzimtico. Depende do fato que o aa C-terminal foi ativado o que favorece a sua ligao ao prximo aa-tRNA

Proofreading translation
Elongation factor-GTP

Translation termination and releasing factors

Molecular mimicry

Polyribosomes

INHIBITOR
Acting only on bacteria Tetracycline Streptomycin Chloramphenicol Erythromycin Rifamycin

SPECIFIC EFFECT

blocks binding of aminoacyl-tRNA to A-site of ribosome prevents the transition from initiation complex to chain-elongating ribosome and also causes miscoding blocks the peptidyl transferase reaction on ribosomes blocks the translocation reaction on ribosomes blocks initiation of RNA chains by binding to RNA polymerase

Acting on bacteria and eucaryotes Puromycin causes the premature release of nascent polypeptide chains by its addition to growing chain end Actinomycin D binds to DNA and blocks the movement of RNA polymerase (prevents RNA synthesis) Acting on eucaryotes but not bacteria Cycloheximide blocks the translocation reaction on ribosomes (step 3 in Figure 6-65) Anisomycin blocks the peptidyl transferase reaction on ribosomes (step 2 in Figure 6-65) -Amanitin blocks mRNA synthesis by binding preferentially to RNA polymerase II
The ribosomes of eucaryotic mitochondria (and chloroplasts) often resemble those of bacteria in their sensitivity to inhibitors. Therefore, some of these antibiotics can have a deleterious effect on human mitochondria.

Codon bias

Accuracy of translation Nonsense-mediated mRNA decay

Ribosomes introduce wrong aa 1/10.000

Protein folding

Abnormally folded proteins aggregate and precipitate

Incorrectly folded proteins precipitate

B-amyloid

Abnormally folded proteins are rapidly degraded