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PDBsum entry 2w2u
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Hydrolase/transport
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PDB id
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2w2u
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References listed in PDB file
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Key reference
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Title
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A role for the escrt system in cell division in archaea.
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Authors
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R.Y.Samson,
T.Obita,
S.M.Freund,
R.L.Williams,
S.D.Bell.
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Ref.
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Science, 2008,
322,
1710-1713.
[DOI no: ]
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PubMed id
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Abstract
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Archaea are prokaryotic organisms that lack endomembrane structures. However, a
number of hyperthermophilic members of the Kingdom Crenarchaea, including
members of the Sulfolobus genus, encode homologs of the eukaryotic endosomal
sorting system components Vps4 and ESCRT-III (endosomal sorting complex required
for transport-III). We found that Sulfolobus ESCRT-III and Vps4 homologs
underwent regulation of their expression during the cell cycle. The proteins
interacted and we established the structural basis of this interaction.
Furthermore, these proteins specifically localized to the mid-cell during cell
division. Overexpression of a catalytically inactive mutant Vps4 in Sulfolobus
resulted in the accumulation of enlarged cells, indicative of failed cell
division. Thus, the archaeal ESCRT system plays a key role in cell division.
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Figure 2.
Fig. 2. Localization of (Top) Saci1372 (Vps4) and (bottom)
Saci1373 (ESCRT-III). Representative images are shown. Images
show the FM4-64X staining for membrane (red), DAPI staining for
DNA (blue), antibody labeling of ESCRT-III or Vps4(green), and
merged images. Scale bar, 1 µm. Additional images are
shown in figs. S6 and S7 and movie S1.
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Figure 3.
Fig. 3. (A) Interactions between ESCRT-III proteins detected by
yeast two-hybrid analyses. Yeast cells containing the indicated
plasmids were plated in media lacking leucine and tryptophan to
select for plasmids and lacking histidine to score for
interactions. (B) Identification of the minimal interaction site
on Saci1373 (ESCRT-III) for binding Vps4. Glutathione
S-transferase (GST) fusions of ESCRT-III fragments were used in
pull-down assays with the full-length Vps4. The results of the
pull-downs are shown in the lower panel. The input lanes contain
25 and 5% of input. (C) Identification of the interaction domain
of Vps4 that binds Saci1373 (ESCRT-III). GST-Saci1373
(ESCRT-III) was used in pull-down assays with the full-length
(1), C-terminal AAA+ domain (2), or isolated MIT domain (3) of
Vps4. (D) A schematic representation of the interaction of
Saci1373 (red) with the Saci1372 Vps4 MIT domain (yellow). (E)
An illustration of the interaction of the yeast Vps4 MIT domain
(yellow) with the C-terminal MIM1 motif (blue) of the yeast Vps2
ESCRT-III subunit. The MIM1 motif slots between MIT helices  2
and 3 (14). (F) The
Saci1373 MIM2 (red)/Saci1372 MIT (yellow) interaction is closely
related in structure with the CHMP6 MIM2 (green, extended)/VPS4A
MIT (gray helices) interaction (20).
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The above figures are
reprinted
by permission from the AAAs:
Science
(2008,
322,
1710-1713)
copyright 2008.
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