In-depth study of DNA binding of Cys2His2 finger domains in testis zinc-finger protein.
Previously, we identified that both fingers 1 and 2 in the three Cys2His2 zinc-finger domains (TZD) of testis zinc-finger protein specifically bind to its cognate DNA; however, finger 3 is non-sequence-specific. To gain insights into the interaction mechanism, here we further investigated the DNA-bi...
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doaj-2b024eb6d2664cc596ba223d664cd6142020-11-24T20:50:15ZengPublic Library of Science (PLoS)PLoS ONE1932-62032017-01-01124e017505110.1371/journal.pone.0175051In-depth study of DNA binding of Cys2His2 finger domains in testis zinc-finger protein.Chun-Chi ChouShu-Yi WeiYuan-Chao LouChinpan ChenPreviously, we identified that both fingers 1 and 2 in the three Cys2His2 zinc-finger domains (TZD) of testis zinc-finger protein specifically bind to its cognate DNA; however, finger 3 is non-sequence-specific. To gain insights into the interaction mechanism, here we further investigated the DNA-binding characteristics of TZD bound to non-specific DNAs and its finger segments bound to cognate DNA. TZD in non-specific DNA binding showed smaller chemical shift perturbations, as expected. However, the direction of shift perturbation, change of DNA imino-proton NMR signal, and dynamics on the 15N backbone atom significantly differed between specific and non-specific binding. Using these unique characteristics, we confirmed that the three single-finger segments (TZD1, TZD2 and TZD3) and the two-finger segment (TZD23) non-specifically bind to the cognate DNA. In comparison, the other two-finger segment (TZD12) binding to the cognate DNA features simultaneous non-specific and semi-specific binding, both slowly exchanged in terms of NMR timescale. The process of TZD binding to the cognate DNA is likely stepwise: initially TZD non-specifically binds to DNA, then fingers 1 and 2 insert cooperatively into the major groove of DNA by semi-specific binding, and finally finger 3 non-specifically binds to DNA, which promotes the specific binding on fingers 1 and 2 and stabilizes the formation of a specific TZD-DNA complex.http://europepmc.org/articles/PMC5383199?pdf=render |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Chun-Chi Chou Shu-Yi Wei Yuan-Chao Lou Chinpan Chen |
spellingShingle |
Chun-Chi Chou Shu-Yi Wei Yuan-Chao Lou Chinpan Chen In-depth study of DNA binding of Cys2His2 finger domains in testis zinc-finger protein. PLoS ONE |
author_facet |
Chun-Chi Chou Shu-Yi Wei Yuan-Chao Lou Chinpan Chen |
author_sort |
Chun-Chi Chou |
title |
In-depth study of DNA binding of Cys2His2 finger domains in testis zinc-finger protein. |
title_short |
In-depth study of DNA binding of Cys2His2 finger domains in testis zinc-finger protein. |
title_full |
In-depth study of DNA binding of Cys2His2 finger domains in testis zinc-finger protein. |
title_fullStr |
In-depth study of DNA binding of Cys2His2 finger domains in testis zinc-finger protein. |
title_full_unstemmed |
In-depth study of DNA binding of Cys2His2 finger domains in testis zinc-finger protein. |
title_sort |
in-depth study of dna binding of cys2his2 finger domains in testis zinc-finger protein. |
publisher |
Public Library of Science (PLoS) |
series |
PLoS ONE |
issn |
1932-6203 |
publishDate |
2017-01-01 |
description |
Previously, we identified that both fingers 1 and 2 in the three Cys2His2 zinc-finger domains (TZD) of testis zinc-finger protein specifically bind to its cognate DNA; however, finger 3 is non-sequence-specific. To gain insights into the interaction mechanism, here we further investigated the DNA-binding characteristics of TZD bound to non-specific DNAs and its finger segments bound to cognate DNA. TZD in non-specific DNA binding showed smaller chemical shift perturbations, as expected. However, the direction of shift perturbation, change of DNA imino-proton NMR signal, and dynamics on the 15N backbone atom significantly differed between specific and non-specific binding. Using these unique characteristics, we confirmed that the three single-finger segments (TZD1, TZD2 and TZD3) and the two-finger segment (TZD23) non-specifically bind to the cognate DNA. In comparison, the other two-finger segment (TZD12) binding to the cognate DNA features simultaneous non-specific and semi-specific binding, both slowly exchanged in terms of NMR timescale. The process of TZD binding to the cognate DNA is likely stepwise: initially TZD non-specifically binds to DNA, then fingers 1 and 2 insert cooperatively into the major groove of DNA by semi-specific binding, and finally finger 3 non-specifically binds to DNA, which promotes the specific binding on fingers 1 and 2 and stabilizes the formation of a specific TZD-DNA complex. |
url |
http://europepmc.org/articles/PMC5383199?pdf=render |
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