New insights into DNA-binding behavior of Wilms tumor protein (WT1)--a dual study.
Nurmemmedov, Elmar LU ; Kimbung, Raymond Yengo LU ; Uysal, Hüseyin LU ; Karlsson, Robert and Thunnissen, Marjolein LU
(2009)
In Biophysical Chemistry
145(2-3).
p.116-125
- Abstract
- Wilms Tumor suppressor protein (WT1) is a transcription factor that is involved in a variety of developmental functions during organ development. It is also implicated in the pathology of several different cancer forms. The protein contains four C(2)H(2)-type zinc fingers and it specifically binds GC-rich sequences in the promoter regions of its target genes, which are either up or down regulated. Two properties make WT1 a more unusual transcription factor - an unconventional amino acid composition for zinc finger 1, and the insertion of a tri-peptide KTS in some of the splice isoforms of WT1. Using six WT1 constructs in which zinc fingers are systematically deleted, a dual study based on a bacterial 1-hybrid system and surface plasmon... (More)
- Wilms Tumor suppressor protein (WT1) is a transcription factor that is involved in a variety of developmental functions during organ development. It is also implicated in the pathology of several different cancer forms. The protein contains four C(2)H(2)-type zinc fingers and it specifically binds GC-rich sequences in the promoter regions of its target genes, which are either up or down regulated. Two properties make WT1 a more unusual transcription factor - an unconventional amino acid composition for zinc finger 1, and the insertion of a tri-peptide KTS in some of the splice isoforms of WT1. Using six WT1 constructs in which zinc fingers are systematically deleted, a dual study based on a bacterial 1-hybrid system and surface plasmon resonance measurements is performed. The experiments show that the effect of zinc finger 1 is not significant in terms of overall DNA-binding kinetics, however it influences both the specificity of target recognition and stability of interaction in presence of KTS. The KTS insertion, however, only mildly retards binding affinity, mainly by affecting the on-rate. We suggest that the insertion disturbs zinc finger 4 from its binding frame, thus weakening the rate of target recognition. Finally, for the construct in which both zinc fingers 1 and 4 were deleted, the two middle fingers 2-3 still could function as a 'minimal DNA-recognition domain' for WT1, however the formation of a stable protein-DNA complex is impaired since the overall affinity was dramatically reduced mainly since the off-rate was severely affected. (Less)
Please use this url to cite or link to this publication:
https://lup.lub.lu.se/record/1500015
- author
- Nurmemmedov, Elmar
LU
; Kimbung, Raymond Yengo
LU
; Uysal, Hüseyin
LU
; Karlsson, Robert
and Thunnissen, Marjolein
LU
- organization
- publishing date
- 2009
- type
- Contribution to journal
- publication status
- published
- subject
- keywords
- Transcription factor, WT1, Zinc finger, Surface plasmon resonance, DNA-binding, Bacterial 1-hybrid system
- in
- Biophysical Chemistry
- volume
- 145
- issue
- 2-3
- pages
- 116 - 125
- publisher
- Elsevier
- external identifiers
-
- wos:000271807500011
- pmid:19853363
- scopus:70350189932
- pmid:19853363
- ISSN
- 1873-4200
- DOI
- 10.1016/j.bpc.2009.09.009
- language
- English
- LU publication?
- yes
- id
- ec7b0cfe-4837-4b2b-9636-f495b86fd316 (old id 1500015)
- date added to LUP
- 2016-04-01 15:07:12
- date last changed
- 2024-10-11 02:07:51
@article{ec7b0cfe-4837-4b2b-9636-f495b86fd316,
abstract = {{Wilms Tumor suppressor protein (WT1) is a transcription factor that is involved in a variety of developmental functions during organ development. It is also implicated in the pathology of several different cancer forms. The protein contains four C(2)H(2)-type zinc fingers and it specifically binds GC-rich sequences in the promoter regions of its target genes, which are either up or down regulated. Two properties make WT1 a more unusual transcription factor - an unconventional amino acid composition for zinc finger 1, and the insertion of a tri-peptide KTS in some of the splice isoforms of WT1. Using six WT1 constructs in which zinc fingers are systematically deleted, a dual study based on a bacterial 1-hybrid system and surface plasmon resonance measurements is performed. The experiments show that the effect of zinc finger 1 is not significant in terms of overall DNA-binding kinetics, however it influences both the specificity of target recognition and stability of interaction in presence of KTS. The KTS insertion, however, only mildly retards binding affinity, mainly by affecting the on-rate. We suggest that the insertion disturbs zinc finger 4 from its binding frame, thus weakening the rate of target recognition. Finally, for the construct in which both zinc fingers 1 and 4 were deleted, the two middle fingers 2-3 still could function as a 'minimal DNA-recognition domain' for WT1, however the formation of a stable protein-DNA complex is impaired since the overall affinity was dramatically reduced mainly since the off-rate was severely affected.}},
author = {{Nurmemmedov, Elmar and Kimbung, Raymond Yengo and Uysal, Hüseyin and Karlsson, Robert and Thunnissen, Marjolein}},
issn = {{1873-4200}},
keywords = {{Transcription factor; WT1; Zinc finger; Surface plasmon resonance; DNA-binding; Bacterial 1-hybrid system}},
language = {{eng}},
number = {{2-3}},
pages = {{116--125}},
publisher = {{Elsevier}},
series = {{Biophysical Chemistry}},
title = {{New insights into DNA-binding behavior of Wilms tumor protein (WT1)--a dual study.}},
url = {{http://dx.doi.org/10.1016/j.bpc.2009.09.009}},
doi = {{10.1016/j.bpc.2009.09.009}},
volume = {{145}},
year = {{2009}},
}