 |
PDBsum entry 1dv0
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
 |
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
DNA binding protein
|
PDB id
|
|
|
|
1dv0
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
 |
Contents |
 |
|
|
|
|
|
|
|
|
* Residue conservation analysis
|
|
|
|
|
References listed in PDB file
|
|
|
Key reference
|
|
|
Title
|
|
Biochemical and structural analysis of the interaction between the uba(2) domain of the DNA repair protein hhr23a and HIV-1 vpr.
|
|
|
Authors
|
|
E.S.Withers-Ward,
T.D.Mueller,
I.S.Chen,
J.Feigon.
|
|
|
Ref.
|
|
Biochemistry, 2000,
39,
14103-14112.
[DOI no: ]
|
|
|
PubMed id
|
|
|
|
|
|
Abstract
|
|
|
The DNA repair protein HHR23A is a highly conserved protein that functions in
nucleotide excision repair. HHR23A contains two ubiquitin associated domains
(UBA) that are conserved in a number of proteins with diverse functions involved
in ubiquitination, UV excision repair, and signaling pathways via protein
kinases. The cellular binding partners of UBA domains remain unclear; however,
we previously found that the HHR23A UBA(2) domain interacts specifically with
the HIV-1 Vpr protein. Analysis of the low resolution solution structure of
HHR23A UBA(2) revealed a hydrophobic loop region of the UBA(2) domain that we
predicted was the interface for protein/protein interactions. Here we present
results of in vitro binding studies that demonstrate the requirement of this
hydrophobic loop region for interaction with human immunodeficiency virus
(HIV-1) Vpr. A single point mutation of the Pro at residue 333 to a Glu totally
abolishes the binding of HIV-1 Vpr to UBA(2). High resolution NMR structures of
the binding deficient UBA(2) mutant P333E as well as of the wild-type UBA(2)
domain were determined to compare the effect of this mutation on the structure.
Small but significant differences are observed only locally at the site of the
mutation. The biochemical and structural analysis confirms the function of the
HHR23A UBA(2) GFP-loop as the protein/protein interacting domain.
|
|
|
Secondary reference #1
|
|
|
Title
|
|
Structure of a human DNA repair protein uba domain that interacts with HIV-1 vpr.
|
|
|
Authors
|
|
T.Dieckmann,
E.S.Withers-Ward,
M.A.Jarosinski,
C.F.Liu,
I.S.Chen,
J.Feigon.
|
|
|
Ref.
|
|
Nat Struct Biol, 1998,
5,
1042-1047.
[DOI no: ]
|
|
|
PubMed id
|
|
|
|
|
|
|
|
|
|
|
|
|
|
Figure 1.
Figure 1. a, Sequence and domain structure of HHR23A. The
different domains that have been identified by sequence analysis
are indicated as shaded boxes. b, Sequences of UBA domains used
in this study.The amino acid positions that are conserved to
more than 90% in all known UBA domains are highlighted.
|
|
Figure 2.
Figure 2. Abrogation of alleviation of Vpr-induced cell cycle
arrest by overexpression of a 134-residue C-terminal portion of
HHR23A, HHR23A(B213trunc), that contains a deletion of the
C-terminal UBA domain. a, Hela cells were co-transfected
with either BSVprThy or pXCThy and a 20-fold molar excess of
either pXC (3.2  g),
pXCB213 (4.0 g),
or pXCB213trunc (4.0 g),
and analyzed by flow cytometry as described^10. Duplicate
cotransfections are shown except for mock-transfected cells.
Panel I, mock transfected cells; panel II, BSVprThy and pXC;
panel III, pXCThy and pXC; panel IV, BSVprThy and pXCB213; panel
V, BSVprThy and pXCB213trunc. b, Expression of HHR23A(B213trunc)
in Hela cells. Hela cells were co-transfected with BSVprThy and
pXCB213 or pXCB213trunc, and analyzed by Western blotting as
described. Lanes 1 and 2 contain lysates from duplicate samples
of cells co-transfected with BSVprThy and pXCB213 (shown in (a )
panel IV), lanes 3 and 4 contain lysates from duplicate samples
of cells co-transfected with BSVprThy and pXCB213trun c (shown
in (a) panel V).
|
|
|
|
|
|
The above figures are
reproduced from the cited reference
with permission from Macmillan Publishers Ltd
|
|
|
|
|
|
|
