Biochimie, volume 171-172, pages 110-123
Analysis of RNA binding properties of human Ku protein reveals its interactions with 7SK snRNA and protein components of 7SK snRNP complex
Shadrina Olga
1
,
Garanina Irina
2
,
Korolev S.
1, 3
,
Zatsepin Timofei S.
1, 4
,
Zatsepin Timofei
,
Van Assche Jeanne
5
,
Daouad Fadoua
5
,
Wallet Clementine
,
Wallet C
5
,
Rohr Olivier
,
Rohr Olivier
5
,
Gottikh Marina B.
1, 3
,
Gottikh Marina
Publication type: Journal Article
Publication date: 2020-04-01
Biochemistry
General Medicine
Abstract
Human Ku heterodimeric protein composed of Ku70 and Ku80 subunits plays an important role in the non-homologous end-joining DNA repair pathway as a sensor of double strand DNA breaks. Ku is also involved in numerous cellular processes, and in some of them it acts in an RNA-dependent manner. However, RNA binding properties of the human Ku have not been well studied. Here we have analyzed interactions of a recombinant Ku heterodimer with a set of RNAs of various structure as well as eCLIP (enhanced crosslinking and immunoprecipitation) data for human Ku70. As a result, we have proposed a consensus RNA structure preferable for the Ku binding that is a hairpin possessing a bulge just near GpG sequence-containing terminal loop. 7SK snRNA is a scaffold for a ribonucleoprotein complex (7SK snRNP), which is known to participate in transcription regulation. We have shown that the recombinant Ku specifically binds a G-rich loop of hairpin 1 within 7SK snRNA. Moreover, Ku protein has been co-precipitated from HEK 293T cells with endogenous 7SK snRNA and such proteins included in 7SK snRNP as HEXIM1, Cdk9 and CTIP2. Ku and Cdk9 binding is found to be RNA-independent, meanwhile HEXIM1 and Ku co-precipitation depended on the presence of intact 7SK snRNA. The latter result has been confirmed using recombinant HEXIM1 and Ku proteins. Colocalization of Ku and CTIP2 was additionally confirmed by confocal microscopy. These results allow us to propose human Ku as a new component of the 7SK snRNP complex.
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Shadrina O. et al. Analysis of RNA binding properties of human Ku protein reveals its interactions with 7SK snRNA and protein components of 7SK snRNP complex // Biochimie. 2020. Vol. 171-172. pp. 110-123.
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Shadrina O., Garanina I., Korolev S., Korolev S., Zatsepin T., Zatsepin T. S., Van Assche J., Daouad F., Wallet C., Wallet C., Rohr O., Rohr O., Gottikh M., Gottikh M. B. Analysis of RNA binding properties of human Ku protein reveals its interactions with 7SK snRNA and protein components of 7SK snRNP complex // Biochimie. 2020. Vol. 171-172. pp. 110-123.
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TY - JOUR
DO - 10.1016/j.biochi.2020.02.016
UR - https://doi.org/10.1016%2Fj.biochi.2020.02.016
TI - Analysis of RNA binding properties of human Ku protein reveals its interactions with 7SK snRNA and protein components of 7SK snRNP complex
T2 - Biochimie
AU - Shadrina, Olga
AU - Garanina, Irina
AU - Korolev, Sergey
AU - Zatsepin, Timofei
AU - Van Assche, Jeanne
AU - Daouad, Fadoua
AU - Wallet, Clementine
AU - Rohr, Olivier
AU - Gottikh, Marina
AU - Korolev, S.
AU - Zatsepin, Timofei S.
AU - Wallet, C
AU - Rohr, Olivier
AU - Gottikh, Marina B.
PY - 2020
DA - 2020/04/01 00:00:00
PB - Elsevier
SP - 110-123
VL - 171-172
SN - 0300-9084
SN - 6183-1638
ER -
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@article{2020_Shadrina,
author = {Olga Shadrina and Irina Garanina and Sergey Korolev and Timofei Zatsepin and Jeanne Van Assche and Fadoua Daouad and Clementine Wallet and Olivier Rohr and Marina Gottikh and S. Korolev and Timofei S. Zatsepin and C Wallet and Olivier Rohr and Marina B. Gottikh},
title = {Analysis of RNA binding properties of human Ku protein reveals its interactions with 7SK snRNA and protein components of 7SK snRNP complex},
journal = {Biochimie},
year = {2020},
volume = {171-172},
publisher = {Elsevier},
month = {apr},
url = {https://doi.org/10.1016%2Fj.biochi.2020.02.016},
pages = {110--123},
doi = {10.1016/j.biochi.2020.02.016}
}
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