Graphics Gallery
Gale Rhodes
Chemistry Department
University of Southern Maine
Revised 2006/08/02
Learn how to use Swiss-PdbViewer. See the Swiss-PdbViewer
Tutorial.
Topic: Translation and the Genetic Code
Examples:
Molecules commonly discussed in biochemistry texts
tRNAPhe
To study the structure of a transfer RNA that carries
phenylalanine (PHE), obtain the file 4TNA
from the Protein Data Bank.
For SwissPdbViewer users, click
here for a prepared view of the 4TNA model, color coded as
follows:
- Amino-acid arm, backbone red in the single-strand region,
green in the double-strand region.
- T-Psi-C arm, backbone cyan.
- DHU arm, backbone yellow.
- Anticodon arm, backbone magenta, with anticodon bases
green.
Explore the structure, and find the following structural
elements:
- The 3'-CCA region and the hydroxyl group to which PHE would be
attached.
- The anticodon region. Use the genetic code table in your text to find
the codons for PHE. Which of the PHE codons would this tRNA recognize?
- What are the orientations of the anticodon bases with respect
to each other? Do you think this orientation allows for matching
with the codon?
- Regions of Watson-Crick double helix.
- Interactions that support the tertiary structure of the
molecule.
Glutaminyl-tRNA Synthetase (A Type-1 Synthetase)
Protein Data Bank file 1QRT
provides a model of glutaminyl-tRNA synthetase bound to
tRNAglu. In addition, ATP is bound at the putative binding
site for the glutaminyl adenylate. Read the Medline abstract linked
to this file.
Swiss-PdbViewer users click here
for a prepared view of the 1QRT model. The model is color coded
as follows:
- tRNA is shown in cyan, with the anticodon region in magenta
backbone with green bases (doesn't look as bad as it sounds -- but
now you know why I ask my wife to approve my clothing before I go
out in public). The 3'-terminal A nucleotide is shown with a
dotted surface, in CPK colors.
- ATP is shown with a dotted surface, in CPK colors.
- Synthetase is shown in ribbon, with helices red and beta
strands yellow. Residues within 4 angstroms of the t-RNA or ATP
are shown fully, CPK colored.
Explore the structure:
- Compute H-bonds and explore the anticodon region of the t-RNA.
Note that recognition of t-RNA by synthetase involves the
anticodon region (residues 34-36, 5'-CUG-3'), but that there are
also other regions of enzyme-tRNA contact (wherever you see enzyme
residues fully displayed (not just ribbon), the enzyme and t-RNA
are in contact).
- Look at the ATP binding region. Measure the distance from the
2'-hydroxyl oxygen of t-RNA residue A76 (the 3'-terminal
nucleotide, which carries the amino acid) to the phosphorus atom
of the middle phosphate of ATP. This phosphorus is at
approximately the position of the aminoacyl carbonyl carbon in the
aminoacyl adenylate that would normally occupy the ATP position
during catalysis.
- Explore the binding of the anticodon bases to the protein.
Compare their relative orientations to those in the anticodon
bases in tRNAPhe, above.
Aspartyl-tRNA Synthetase (A Type-II Synthetase)
Protein Data Bank file 1ASZ
proides a model of aspartyl-tRNA synthetase bound to
tRNAasp. In addition, ATP is aboutn at the putative
binding site for aspartyl adenylate. Read the Medline abstract linked
to this file.
Swiss-PdbViewer users click
here for a prepared view of 1ASZ that allows comparision of the
rRNAasp conformation with that of free tRNAasp
(PDB 2TRA).
Only one of the two complete models from 1ASZ is shown, but both
models are present in the file.The displayed model is color coded as
follows:
- In the 1ASZ layer, tRNA is shown in cyan, with the anticodon
region in magenta backbone with green bases. The 3'-terminal CCA
nucleotides are shown with a dotted surface, in CPK colors.
- ATP is shown with a dotted surface, in CPK colors.
- Synthetase is shown in ribbon, with helices red and beta
strands yellow. Residues within 4 angstroms of the tRNA are shown
fully, CPK colored.
- In the 2TRA layer, the color coding of tRNA is that same. The
3'-CA dinucleotide is missing from this model.
Explore the structure:
- First, using blinking (press tab while holding down
ctrl) to compare the conformations for free and bound
tRNAasp(layers 2TRA and 1ASZ). Give a general
description of the differences between the conformations.
- In layer 1ASZ, compute H-bonds and explore the anticodon
region of the t-RNA. Note that recognition of t-RNA by synthetase
involves the anticodon region, but that there are also other
regions of enzyme-tRNA contact (wherever you see enzyme residues
fully displayed (not just ribbon), the enzyme and t-RNA are in
contact).
- Look at the ATP binding region. Measure the distance from the
2'-hydroxyl oxygen of t-RNA residue A76 (the 3'-terminal
nucleotide, which carries the amino acid) to the phosphorus atom
of the middle phosphate of ATP. This phosphorus is at
approximately the position of the aminoacyl carbonyl carbon in the
aminoacyl adenylate that would normally occupy the ATP position
during catalysis.
- Explore the binding of the anticodon bases (magenta backbone,
green bases) to the protein.
- Blink to compare the stacked orientations of these bases in
free tRNAasp to their orientations in bound
tRNAasp.
- Display the full dimer of 1ASZ (Select: All,
<return>). Show the entire model in ribbons. The tRNA you
have been exploring is green and the other is red.
- Use Select: Groups Close to Another Chain to explore
interactions between the two enzyme subunits.
Does EF-G Mimic EF-Tu/tRNA?
Swiss-PdbViewer users click
here for a project file comparing elongation factor G (EF-G, PDB
2EFG) with a complex of elongation factor Tu (EF-Tu, PDB 1B23).
In the EF-G model, domains 1 and 2 are shown as ribbons, while
domains 3, 4, and 5 are shown in full. In EF-Tu/tRNA, EF-Tu is shown
in ribbon, while tRNA is shown in full. The tRNA is superimposed (by
eye) on domains 3, 4, and 5 of EF-G.
- Blink between the models (hold down ctrl and press
tab repeatedly. Do the last 3 domains of EF-G have a shape
similar to that of the tRNA?
- How might you use Swiss-PdbViewer's tools to strengthen the
case that EF-G might mimic EF-Tu/tRNA?
- If this mimicry does occur, propose a sequence of events
involving EF-G that results in translocation. Remember that an
EF-G-bound GTP is hydrolyzed during this process.
- Suggest a means of testing your proposal.
At each step of translation, a complex of EF-G and GTP is
necessary for translocation of the ribosome to the next codon.
Translocation moves the peptidyl-tRNA from the A site to the P-site.
Next, a complex of EF-Tu and an aminoacyl-tRNA enters the A site,
bringing the next aminoacyl group to be added to the growing peptide
chain. Comparison of the two structures in this project file suggests
that EF-G might mimic the EF-Tu/tRNA complex and displace it
from the A site to the P site.
Topics List
Biochemistry
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