Protein Synthesis: Translation

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Translation

Back 1

process of protein synthesis from an RNA template

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Codon

Back 2

3 bases

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Start codon in mRNA

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AUG (methionine)

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Reading direction

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5' to 3'

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Stop codons

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UAA - you are away

UAG - you are gone

UGA - you go away

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insertion or deletion of nucleotide

Back 6

Can result in frameshift mutation

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where do Modifications take place?

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Everywhere in eukaryotas

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codon - anticodon interaction

Back 8

Flexible than most base pairing

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Wobble

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third position in the codon can change without changing the AA in some cases

allows one tRNA to translate more than one codon

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codon - anticodon pairing

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anticodon

(3') 3-2-1(wobble) (5')

----------

(5') 1-2-3(wobble) (3')

Codon

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Saber leer genetic code

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Types of Mutations

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1. point mutation

2. silent mutation

3. Missense mutation

4. Non sense mutation

5. Insertion (frameshift)

6. Deletion (frameshift)

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point mutation

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changing single nucleotide base

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silent mutation

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Change that specifies the same AA

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Missense mutation

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Change that specifies a different AA

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Non sense mutation

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Change that produces a Stop codon

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Insertion (frameshift)

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Addition of 1 or more bases

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Deletion (frameshift)

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Loss of 1 or more bases

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Components required for protein synthesis

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1. AA

2. tRNA

3. Ribosomes

4. mRNA

5. Aminoacyl-tRNA synthetase

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Aminoacyl-tRNA synthetase

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Catalyses the attachment of AA with its corresponding tRNA forming aminoacyl tRNA

there are 20 synthetases - 1 for each AA

-some have proofreading activity (there recognized and correct)

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tRNA function

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form a covalent bond with a particular AA

base pairing with a codon in mRNA

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tRNA nucleotidyltransferase

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an RNA polymerase that adds CCA sequence to the 3' of humans. It can synthesize a sequence with a template.

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rRNA

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Most abundant and stable RNA's

Are proccessed in the nucleolus

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Ribosome

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Subcellular ribonucleoprotein complex on which protein synthesis occurs

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Cytoplasmic ribosomes in eukaryotic have 4 types of rRNA molecules

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60S --- 5S + 5.8S +28S

40S --- 18S

___________

80S

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Prokaryotic ribosomes have 3 rRNA molecules

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50S --- 5S + 23S

30S --- 16S

________

70S

*mitochondrial are similar

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28S rRNA eukaryotes

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only one with enzymatic activity : peptidyltransferase activity

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Aminoacyl tRNA formation

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in the cytosol (not in the ribosome)

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Translation Initiation complex

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1. 40S and 60S units

2. mRNA

3. initiation factors (elFs)

4. Methionyl-tRNA (only tRNA recognized by the initiation factor elF-2

5. ATP and GTP

6. Kozak consensus sequence

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Reading direction of translation

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5' to 3'

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Initiation in Prokaryotes

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Elongation sites

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E - exit

P - peptidol tRNA (formation of peptide bond)

A - entry site (Aminoacyl- tRNA)

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Elongation

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only use GTP not ATP

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A site in Elongation

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GTP is hydrolized to GDP

the binding comprises the second error checking

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Peptidyltransferase

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catalyze the peptide bond between A and P site

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GTP used in translation

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Initiation: 1 GTP

Elongation: **

Termination: 1GTP

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ATP used in translation

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only in initiation

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polysome

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complez of ribosomes that translate at the same time

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chaperones

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Prevent improper interactions from occurring in the poly peptide chain

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targeting sequences or signal sequences

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Amino acid sequences that determine where the proteins will be released

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Synthesis of protein in the RER

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Golgi complex

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***

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Tetracycline

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Binds to the 30S subunit and inhibits binding ofaminoacyl-tRNA to the A site

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Cyclohexamide

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Inhibits the peptidyltransferase activity of the60S ribosomal subunit (eukaryotes)

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Chloramphenicol

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Binds to the 50S ribosomal subunit andinhibits peptidyltransferase

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Erythromycin

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Binds to the 50S ribosomal subunit and preventstranslocation

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Puromycin

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Causes premature chain termination by acting as aanalog of aminoacyl-tRNA (both)

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Streptomycin

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Binds to the 30S ribosomal subunit ofprokaryotes, thereby preventing formation of the initiationcomplex. It also causes misreading of mRNA

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Diphtheria Pertusis Toxin

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Secretes the A fragment (toxin) a ADP ribosyl transferase

eEF-2 is inhibited - thus protein synthesis stops

inactivation results in cell death

Vaccination will prevent it

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Tay-Sachs Disease

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Ganglioside (Gm2) accumulates
because B-hexosaminidase (in lysosome) cannot catalyze Gm2 to Gm3.

Mutation in gene encoding lysosomal enzyme

this mutation lost of lysosomal function and death at an early age

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I cell Disease

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inherited lysosomal storage disorder

Mutation post translational

-mistargeting of enzymes to lysosomes

In GNTPA gene

coarse facil features, skeletal abnormalities and mental retardation