NucleaRDB: Extraction of mutation data from the literature

2005, NucleaRDB.

This data was extracted from Medline abstracts and full texts (when available) in an automated manner.

The table below describes the selected point mutation and provides links to other documents. The sentence(s) where the point mutation G21E was found are listed after the table.

The mutated residues are also indicated in the family sequence alignments and hyperlinked to the corresponding mutation pages.

Point mutation G21E in ANDR_HUMAN

Point mutation:	G21E
Domain:	N-term
General numbering (NucleaRDB):	-
Protein:	ANDR_HUMAN (AR,DHTR, NR3C)	Swiss-Prot Cross-reference table Family page
Other point mutations / same protein	List of mutations in ANDR_HUMAN
Family alignments	3C4 Androgen (AR) 3C Glucocorticoid-like (GR,MR,PR,AR) 3 Estrogen like (ER,ERR,GR,MR,PR,AR)
Other point mutations / same position
Reference:	Dual function of an amino-terminal amphipatic helix in androgen receptor-mediated transactivation through specific and nonspecific response elements. Callewaert L, Verrijdt G, Christiaens V, Haelens A, Claessens F J Biol Chem 2003 Mar 7;278(10):8212-8.	Medline
Other point mutations / same article	List
Text source	HTML and PDF full texts
Validation status	True positive

Relevant sentences:

G21E

Deletion of a FQNLF core sequence in this region blunts the interaction , as does a G21E mutation
Although the G21E mutation strongly impairs the amino / carboxyl-terminal interaction , it does not significantly influence androgen receptor activity on either selective or nonselective elements
The pSG5 construct expressing the G21E-mutated hAR was generated by coincidence and was detected by sequencing
The second construct has a point mutation immediately amino-terminally of the FQNLF motif ; Gly21 is mutated to a Glu (AR1-529G21E)
Surprisingly , the G21E mutation in the NTD , outside the FQNLF motif , also markedly reduces N / C interaction
A , schematical representation of the amino-terminal mutated domains Delta FQNLF , G21E , and L435P fused to the VP16 transactivation domain
Analysis of Gly21 Flanking the FQNLF Core-- The point mutation G21E shows a decreased AR N / C interaction similar to the deletion of the FQNLF core in a mammalian two-hybrid assay (Fig. 1 C)
Surprisingly , the transcriptional activation of the TAT-GRE construct is decreased only marginally when the G21E mutation is introduced in the AR , and no clear effect is seen when the androgen-selective slp-HRE2 was tested
When the nonselective mutant of slp-HRE2 was used , the G21E mutation again had no effect on the functionality of the AR (Fig. 5 )
Transcriptional activation of the wtAR and ARG21E at specific versus nonspecific HREs
COS-7 cells were transfected with 100 ng of luciferase reporter constructs containing two copies of the response elements indicated on the left and cotransfected with 20 ng of empty vector , pSG5wtAR , or pSG5ARG21E
Effects of Mutations within the NTD on DNA Binding-- To analyze the influence of the deletion of the FQNLF motif and the G21E and the L435P mutations on the in vitro interaction of the full size receptor with nonspecific and specific AREs , band shifts assays using wtAR , ARDelta FQNLF , ARG21E , and ARL435P were performed (Fig. 6 A)
COS-7 cells were transfected with expression vectors for wtAR , ARDelta FQNLF , ARG21E , and ARL435P (1 µg)
A , gel shift assays of the TAT-GRE , slp-HRE2 and slp-HRE2 mut (-4T-A ; +2A-T) HREs with wtAR , ARDelta FQNLF , ARG21E , and ARL435P
Labeled probes (indicated at the top of the panels) were incubated with 5 µl of COS-7 extracts containing wtAR , ARDelta FQNLF , ARG21E , or ARL435P (lanes 2 and 3 , lanes 4 and 5 , lanes 6 and 7 , and lanes 8 and 9 , respectively)
Equal amounts of COS extracts containing ARDelta FQNLF , ARG21E , and ARL435P as used in the supershift assay and as indicated at the top were used as to perform Western blot
Deletion of FQNLF and the G21E mutation resulted in an activation comparable with that of wtAR-NTD , whereas the L435P mutation resulted in a decreased activation
pABGal4-DBDQrSRC-1e (989-1240) (50 ng / well) was coexpressed in COS-7 cells with 50 ng of empty pSNATCH-II , pSNATCH-IIAR1-529 , pSNATCH-IIAR1-529Delta FQNLF , pSNATCH-IIAR1-529G21E , or pSNATCH-IIAR1-529L435P
COS-7 cells were transfected with 20 ng of empty vector and pSG5 with wtAR , ARDelta FQNLF , ARG21E , or ARL435P and cotransfected with 100 ng pGEM-T or pSG5SRC-1e
We tested whether the mutations AR1-529Delta FQNLF , AR1-529G21E , and AR1-529L435P , fused to the VP16 activation domain , are still able to interact with the Qr of SRC-1 , fused to the Gal4-DBD (Fig. 7 B)
A striking characteristic for the point mutation G21E is that , although the NTD shows a strongly reduced interaction with the AR-LBD (Fig. 1 C) , there is a 2.5-fold better interaction with Qr when compared with wtAR-NTD
To compare the interaction of Qr with AR-NTD and its mutants with the coactivation ability of SRC-1e , the constructs pSG5 , pSG5wtAR , pSG5Delta FQNLF , pSG5G21E , and pSG5L435P and the 2×TAT-GRE(E1b) luciferase reporter construct were transiently transfected into COS-7 cells and cotransfected with pGEM-T or pSG5SRC-1e (Fig. 7 C)
Transcriptional activity of ARG21E and ARL435P increased to the same level compared with wtAR in the presence of SRC-1e , whereas the net coactivation of ARDelta FQNLF in the presence of SRC-1e is lower
In this study , we compared the transcriptional activity of the constructs ARDelta FQNLF , ARG21E , and ARL435P on specific versus nonspecific response elements and enhancers (Figs
The G21E mutation strongly reduces N / C interaction (Fig. 1 C)
This might be explained by the residual AR-NTD G21E / AR-LBD interaction , observed in Fig. 1 C , which could be sufficient for AR activity on the nonspecific HREs
A second interesting feature for the G21E mutation is an almost 3-fold increase in the affinity for the Qr of SRC-1 as measured in double-hybrid assays (Fig. 7 B)
It should be noted that deletion of the FQNLF core and the G21E mutation does not affect the intrinsic activity of the NTD , in contrast to L435P mutation , which reduces the activity 2-fold (Fig. 1 C)
Possibly , the lowered affinity of the NTDG21E for the LBD and its effect on transactivation by the AR is compensated by the increased affinity of the NTD for the Qr of SRC-1
SRC-1e coexpression seems to rescue the ARG21E activity to the same level as wtAR and ARL435P (Fig. 7 C)
However , the effects of the G21E mutation on Qr recruitment by the isolated NTD (Fig. 7 B) leads us to propose a more direct role in the p160 recruitment
A , schematical representation of the amino-terminal mutated domains FQNLF , G21E , and L435P fused to the VP16 transactivation domain
Effects of Mutations within the NTD on DNA Binding--To analyze the influence of the deletion of the FQNLF motif and the G21E and the L435P mutations on the in vitro interaction of the full size receptor with nonspecific and specific AREs , band shifts assays using wtAR , AR FQNLF , ARG21E , and ARL435P were performed (Fig. 6A)
COS-7 cells were transfected with expression vectors for wtAR , AR FQNLF , ARG21E , and ARL435P (1 g)
We tested whether the mutations AR1529 FQNLF , AR1529G21E , and AR1529L435P , fused to the VP16 activation domain , are still able to interact with the Qr of SRC-1 , fused to the Gal4-DBD (Fig. 7B)
To compare the interaction of Qr with AR-NTD and its mutants with the coactivation ability of SRC-1e , the constructs pSG5 , pSG5wtAR , pSG5 FQNLF , pSG5G21E , and pSG5L435P and the 2 TAT-GRE(E1b) luciferase reporter construct were transiently transfected into COS-7 cells and cotransfected with pGEM-T or pSG5SRC-1e (Fig. 7C)
Transcriptional activity of ARG21E and ARL435P increased to the same level compared with wtAR in the presence of SRC-1e , whereas
FIG

A , gel shift assays of the TAT-GRE , slp-HRE2 and slp-HRE2 mut ( 4T-A ; 2A-T) HREs with wtAR , AR FQNLF , ARG21E , and ARL435P

Labeled probes (indicated at the top of the panels) were incubated with 5 l of COS-7 extracts containing wtAR , AR FQNLF , ARG21E , or ARL435P (lanes 2 and 3 , lanes 4 and 5 , lanes 6 and 7 , and lanes 8 and 9 , respectively)

Equal amounts of COS extracts containing AR FQNLF , ARG21E , and ARL435P as used in the supershift assay and as indicated at the top were used as to perform Western blot

pABGal4-DBDQrSRC-1e (989 1240) (50 ng / well) was coexpressed in COS-7 cells with 50 ng of empty pSNATCH-II , pSNATCH-IIAR1529 , pSNATCH-IIAR1529 FQNLF , pSNATCH-IIAR1529G21E , or pSNATCH-IIAR1529L435P

COS-7 cells were transfected with 20 ng of empty vector and pSG5 with wtAR , AR FQNLF , ARG21E , or ARL435P and cotransfected with 100 ng pGEM-T or pSG5SRC-1e

In this study , we compared the transcriptional activity of the constructs AR FQNLF , ARG21E , and ARL435P on specific versus nonspecific response elements and enhancers (Figs

F.Horn (nucleardbcmbi.kun.nl), 21-Apr-2005