Concept 17.4: Translation is the RNA-directed synthesis of a polypeptide: a closer look
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1 Concept 17.4: Translation is the RNA-directed synthesis of a polypeptide: a closer look A cell translates an message into protein with the help of transfer RNA () Molecules of are not identical: ach carries a specific amino acid on one end ach has an anticodon on the other end; the anticodon base-pairs with a complementary codon on Fig Polypeptide Amino acids Ribosome with amino acid attached Anticodon Codons 1
2 The Structure and Function of Transfer RNA A molecule consists of a single RNA strand that is only about 80 nucleotides long Flattened into one plane to reveal its base pairing, a molecule looks like a cloverleaf Because of hydrogen bonds, actually twists and folds into a three-dimensional molecule is roughly L-shaped A C C Fig Amino acid attachment site Hydrogen bonds Anticodon (a) Two-dimensional structure Amino acid attachment site Hydrogen bonds Anticodon (b) Three-dimensional structure Anticodon (c) Symbol used in this book 2
3 Accurate translation requires two steps: First: a correct match between a and an amino acid, done by the enzyme aminoacyl synthetase Second: a correct match between the anticodon and an codon Flexible pairing at the third base of a codon is called wobble and allows some s to bind to more than one codon Fig Amino acid Aminoacyl- synthetase (enzyme) P P P Adenosine ATP 3
4 Fig Amino acid Aminoacyl- synthetase (enzyme) P P P Adenosine ATP P Adenosine P P i P i P i Fig Amino acid Aminoacyl- synthetase (enzyme) P P P Adenosine ATP denosine P P i P i P i Aminoacyl- synthetase P Adenosine AMP Computer model 4
5 Fig Amino acid Aminoacyl- synthetase (enzyme) P P P Adenosine ATP denosine P P i P i P i Aminoacyl- synthetase P Adenosine AMP Computer model Aminoacyl- ( charged ) Ribosomes Ribosomes facilitate specific coupling of anticodons with codons in protein synthesis The two ribosomal s (large and small) are made of proteins and ribosomal RNA (rrna) 5
6 Fig molecules Growing polypeptide xit tunnel Large Small (a) Computer model of functioning ribosome P site (Peptidyl- binding site) site (xit site) binding site A site (Aminoacyl binding site) Large Small (b) Schematic model showing binding sites Amino end Growing polypeptide Next amino acid to be added to polypeptide chain Codons (c) Schematic model with and Fig a molecules Growing polypeptide xit tunnel Large Small (a) Computer model of functioning ribosome 6
7 Fig b P site (Peptidyl- binding site) site (xit site) binding site A site (Aminoacyl binding site) Large Small (b) Schematic model showing binding sites Amino end Codons Growing polypeptide Next amino acid to be added to polypeptide chain (c) Schematic model with and A ribosome has three binding sites for : The A site holds the that carries the next amino acid to be added to the chain The P site holds the that carries the growing polypeptide chain The site is the exit site, where discharged s leave the ribosome 7
8 Translation Stage 1: Ribosome Association and Initiation of Translation The initiation stage of translation brings together, a with the first amino acid, and the two ribosomal s First, a small ribosomal binds with and a special initiator Then the small moves along the until it reaches the start codon on (5 - AUG-3 ) Proteins called initiation factors bring in the large that completes the translation initiation complex Fig U A C A U G P site Large ribosomal Initiator Start codon binding site Small ribosomal GTP GDP A Translation initiation complex 8
9 Translation Stage 2: longation of the Polypeptide Chain During the elongation stage, amino acids are added one by one to the preceding amino acid ach addition involves proteins called elongation factors and occurs in three steps: codon recognition, peptide bond formation, and translocation Fig Amino end of polypeptide site site 9
10 Fig Amino end of polypeptide site site GTP GDP Fig Amino end of polypeptide site site GTP GDP 10
11 Fig Amino end of polypeptide Ribosome ready for next aminoacyl site site GTP GDP GDP GTP Translation Stage 3: Termination of Translation Termination occurs when a stop codon in the reaches the A site of the ribosome The A site accepts a protein called a release factor The release factor causes the addition of a water molecule instead of an amino acid This reaction releases the polypeptide, and the translation assembly then comes apart 11
12 Fig Release factor Stop codon (UAG, UAA, or UGA) Fig Release factor Free polypeptide Stop codon (UAG, UAA, or UGA) 2 GTP 2 GDP 12
13 Fig Release factor Free polypeptide Stop codon (UAG, UAA, or UGA) 2 GTP 2 GDP Polyribosomes A number of ribosomes can translate a single simultaneously, forming a polyribosome (or polysome) Polyribosomes enable a cell to make many copies of a polypeptide very quickly 13
14 Fig Growing polypeptides Completed polypeptide Incoming ribosomal s (a) Start of ( end) nd of ( end) Ribosomes (b) 0.1 µm Protein Folding and Post-Translational Modifications During and after synthesis, a polypeptide chain spontaneously coils and folds into its threedimensional shape Proteins may also require post-translational modifications before doing their job Some polypeptides are activated by enzymes that cleave them Other polypeptides come together to form the s of a protein 14
15 Targeting Polypeptides to Specific Locations Two populations of ribosomes are evident in cells: free ribsomes (in the cytoplasm) and bound ribosomes (attached to the R) Free ribosomes mostly synthesize proteins that function in the cytoplasm Bound ribosomes make proteins of the endomembrane system and proteins that are secreted from the cell Ribosomes are identical and can switch from free to bound Polypeptide synthesis always begins in the cytosol Synthesis finishes in the cytosol unless the polypeptide signals the ribosome to attach to the R Polypeptides destined for the R or for secretion are marked by a signal peptide 15
16 A signal-recognition particle (SRP) binds to the signal peptide The SRP brings the signal peptide and its ribosome to the R Fig Ribosome Signal peptide Signalrecognition particle (SRP) CYTOSOL R LUMN SRP receptor protein Translocation complex Signal peptide removed R membrane Protein What happens to a protein when the cell doesn t need it anymore? (We ll talk more about this later.) 16
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