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PDBsum entry 1vs7
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Contents |
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206 a.a.
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205 a.a.
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150 a.a.
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100 a.a.
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152 a.a.
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129 a.a.
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127 a.a.
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98 a.a.
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117 a.a.
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123 a.a.
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113 a.a.
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96 a.a.
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88 a.a.
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80 a.a.
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81 a.a.
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55 a.a.
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80 a.a.
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85 a.a.
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218 a.a.
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51 a.a.
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References listed in PDB file
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Key reference
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Title
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Structural analysis of kasugamycin inhibition of translation.
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Authors
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B.S.Schuwirth,
J.M.Day,
C.W.Hau,
G.R.Janssen,
A.E.Dahlberg,
J.H.Cate,
A.Vila-Sanjurjo.
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Ref.
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Nat Struct Mol Biol, 2006,
13,
879-886.
[DOI no: ]
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PubMed id
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Abstract
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The prokaryotic ribosome is an important target of antibiotic action. We
determined the X-ray structure of the aminoglycoside kasugamycin (Ksg) in
complex with the Escherichia coli 70S ribosome at 3.5-A resolution. The
structure reveals that the drug binds within the messenger RNA channel of the
30S subunit between the universally conserved G926 and A794 nucleotides in 16S
ribosomal RNA, which are sites of Ksg resistance. To our surprise, Ksg
resistance mutations do not inhibit binding of the drug to the ribosome. The
present structural and biochemical results indicate that inhibition by Ksg and
Ksg resistance are closely linked to the structure of the mRNA at the junction
of the peptidyl-tRNA and exit-tRNA sites (P and E sites).
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Figure 2.
Figure 2. Determinants of resistance to Ksg in the DASL. (a)
Location of the DASL relative to Ksg. DASL residues A1518 and
A1519 are shown (gold). Also shown are A792 and G1497 (gray),
Ksg (cyan) and distances from the DASL to these residues (in
Angstroms). (b) Post-transcriptional modification of the DASL.
Plasmid-encoded 16S rRNAs carrying mutations in the DASL (or
wild-type (WT) rRNAs) were isolated from ksgA^+ (lanes 3, 7, 11,
15, 19 and 23) or ksgA^- strains (lanes 4, 8, 12, 16, 20 and
24). The presence of the methyl groups at A1518 and/or A1519 in
16S rRNA is detected as a strong reverse-transcriptase stop in
primer extension^19. Sequencing lanes, containing rRNA isolated
from the ksgA^- strain, are included for every mutant.
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Figure 3.
Figure 3. Chemical modification of mutant and wild-type 30S
subunits in the presence of Ksg. Sequencing lanes are labeled
U, G, C and A. (a) Ksg footprint in the 794 region of the 16S
rRNA upon chemical modification with DMS. (b) Ksg footprint in
the 926 region of the 16S rRNA upon chemical modification with
kethoxal. The A794G mutation renders the DMS reactivity of this
base undetectable, and the G926A mutation makes this residue
unreactive to chemical attack by kethoxal; thus, Ksg binding to
ribosomes carrying a mutation at one residue was detected as
protection of the other residue. Ksg was used at 0, 64 and 320
 g
ml^-1 (triangles denote increasing concentration). WT, wild-type
rRNA.
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The above figures are
reprinted
by permission from Macmillan Publishers Ltd:
Nat Struct Mol Biol
(2006,
13,
879-886)
copyright 2006.
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