Single-molecule visualization of human RECQ5 interactions with
single-stranded DNA recombination intermediates

J 2021

Single-molecule visualization of human RECQ5 interactions with single-stranded DNA recombination intermediates

XUE, C. Y., Lucia MOLNÁROVÁ, J. B. STEINFELD, W. X. ZHAO, C. J. MA et. al.

Basic information

Original name

Single-molecule visualization of human RECQ5 interactions with single-stranded DNA recombination intermediates

Authors

XUE, C. Y., Lucia MOLNÁROVÁ (703 Slovakia, belonging to the institution), J. B. STEINFELD, W. X. ZHAO, C. J. MA, Mário ŠPÍREK (703 Slovakia, belonging to the institution), K. KANIECKI, Y. KWON, Ondrej BELAN (203 Czech Republic), Kateřina KREJČÍ (203 Czech Republic, belonging to the institution), S. J. BOULTON, P. SUNG, E. C. GREENE and Lumír KREJČÍ (203 Czech Republic, guarantor, belonging to the institution)

Edition

Nucleic acids research, Oxford, Oxford University Press, 2021, 0305-1048

Other information

Language

English

Type of outcome

Článek v odborném periodiku

Field of Study

10608 Biochemistry and molecular biology

Country of publisher

United Kingdom of Great Britain and Northern Ireland

Confidentiality degree

není předmětem státního či obchodního tajemství

References:

URL

Impact factor

Impact factor: 19.160

RIV identification code

RIV/00216224:14110/21:00118917

Organization unit

Faculty of Medicine

DOI

http://dx.doi.org/10.1093/nar/gkaa1184

UT WoS

000610552100027

Keywords in English

REPLICATION PROTEIN-A; HOMOLOGOUS RECOMBINATION; BLOOMS-SYNDROME; RAD51 FILAMENTS; ATP HYDROLYSIS; POLYMERASE-II; HRDC DOMAIN; HELICASE; SRS2; REPAIR

Abstract

V originále

RECQ5 is one of five RecQ helicases found in humans and is thought to participate in homologous DNA recombination by acting as a negative regulator of the recombinase protein RAD51. Here, we use kinetic and single molecule imaging methods to monitor RECQ5 behavior on various nucleoprotein complexes. Our data demonstrate that RECQ5 can act as an ATP-dependent single-stranded DNA (ssDNA) motor protein and can translocate on ssDNA that is bound by replication protein A (RPA). RECQ5 can also translocate on RAD51-coated ssDNA and readily dismantles RAD51-ssDNA filaments. RECQ5 interacts with RAD51 through protein-protein contacts, and disruption of this interface through a RECQ5-F666A mutation reduces translocation velocity by similar to 50%. However, RECQ5 readily removes the ATP hydrolysis-deficient mutant RAD51-K133R from ssDNA, suggesting that filament disruption is not coupled to the RAD51 ATP hydrolysis cycle. RECQ5 also readily removes RAD51-I287T, a RAD51 mutant with enhanced ssDNA-binding activity, from ssDNA. Surprisingly, RECQ5 can bind to double-stranded DNA (dsDNA), but it is unable to translocate. Similarly, RECQ5 cannot dismantle RAD51-bound heteroduplex joint molecules. Our results suggest that the roles of RECQ5 in genome maintenance may be regulated in part at the level of substrate specificity.

Links

EF16_025/0007381, research and development project

Name: Preklinická progrese nových organických sloučenin s cílenou biologickou aktivitou

EF16_027/0008360, research and development project

Name: Postdoc@MUNI

GA17-17720S, research and development project

Name: Vnitřní vlastnosti RAD51 vlákna a jeho biologické regulace

Investor: Czech Science Foundation

206292/E/17/Z, interní kód MU

Name: Mechanics and execution of homologous recombination - biophysics to the organism

Investor: Wellcome Trust

Citovat

XUE, C. Y., Lucia MOLNÁROVÁ, J. B. STEINFELD, W. X. ZHAO, C. J. MA, Mário ŠPÍREK, K. KANIECKI, Y. KWON, Ondrej BELAN, Kateřina KREJČÍ, S. J. BOULTON, P. SUNG, E. C. GREENE and Lumír KREJČÍ. Single-molecule visualization of human RECQ5 interactions with single-stranded DNA recombination intermediates. Nucleic acids research. Oxford: Oxford University Press, 2021, vol. 49, No 1, p. 285-305. ISSN 0305-1048. Available from: https://dx.doi.org/10.1093/nar/gkaa1184.

@article{1758048,
   author = {Xue, C. Y. and Molnárová, Lucia and Steinfeld, J. B. and Zhao, W. X. and Ma, C. J. and Špírek, Mário and Kaniecki, K. and Kwon, Y. and Belan, Ondrej and Krejčí, Kateřina and Boulton, S. J. and Sung, P. and Greene, E. C. and Krejčí, Lumír},
   article_location = {Oxford},
   article_number = {1},
   doi = {http://dx.doi.org/10.1093/nar/gkaa1184},
   keywords = {REPLICATION PROTEIN-A; HOMOLOGOUS RECOMBINATION; BLOOMS-SYNDROME; RAD51 FILAMENTS; ATP HYDROLYSIS; POLYMERASE-II; HRDC DOMAIN; HELICASE; SRS2; REPAIR},
   language = {eng},
   issn = {0305-1048},
   journal = {Nucleic acids research},
   title = {Single-molecule visualization of human RECQ5 interactions with single-stranded DNA recombination intermediates},
   url = {https://academic.oup.com/nar/article/49/1/285/6041011},
   volume = {49},
   year = {2021}
}

TY  - JOUR
ID  - 1758048
AU  - Xue, C. Y. - Molnárová, Lucia - Steinfeld, J. B. - Zhao, W. X. - Ma, C. J. - Špírek, Mário - Kaniecki, K. - Kwon, Y. - Belan, Ondrej - Krejčí, Kateřina - Boulton, S. J. - Sung, P. - Greene, E. C. - Krejčí, Lumír
PY  - 2021
TI  - Single-molecule visualization of human RECQ5 interactions with single-stranded DNA recombination intermediates
JF  - Nucleic acids research
VL  - 49
IS  - 1
SP  - 285-305
EP  - 285-305
PB  - Oxford University Press
SN  - 03051048
KW  - REPLICATION PROTEIN-A
KW  - HOMOLOGOUS RECOMBINATION
KW  - BLOOMS-SYNDROME
KW  - RAD51 FILAMENTS
KW  - ATP HYDROLYSIS
KW  - POLYMERASE-II
KW  - HRDC DOMAIN
KW  - HELICASE
KW  - SRS2
KW  - REPAIR
UR  - https://academic.oup.com/nar/article/49/1/285/6041011
N2  - RECQ5 is one of five RecQ helicases found in humans and is thought to participate in homologous DNA recombination by acting as a negative regulator of the recombinase protein RAD51. Here, we use kinetic and single molecule imaging methods to monitor RECQ5 behavior on various nucleoprotein complexes. Our data demonstrate that RECQ5 can act as an ATP-dependent single-stranded DNA (ssDNA) motor protein and can translocate on ssDNA that is bound by replication protein A (RPA). RECQ5 can also translocate on RAD51-coated ssDNA and readily dismantles RAD51-ssDNA filaments. RECQ5 interacts with RAD51 through protein-protein contacts, and disruption of this interface through a RECQ5-F666A mutation reduces translocation velocity by similar to 50%. However, RECQ5 readily removes the ATP hydrolysis-deficient mutant RAD51-K133R from ssDNA, suggesting that filament disruption is not coupled to the RAD51 ATP hydrolysis cycle. RECQ5 also readily removes RAD51-I287T, a RAD51 mutant with enhanced ssDNA-binding activity, from ssDNA. Surprisingly, RECQ5 can bind to double-stranded DNA (dsDNA), but it is unable to translocate. Similarly, RECQ5 cannot dismantle RAD51-bound heteroduplex joint molecules. Our results suggest that the roles of RECQ5 in genome maintenance may be regulated in part at the level of substrate specificity.
ER  -

XUE, C. Y., Lucia MOLNÁROVÁ, J. B. STEINFELD, W. X. ZHAO, C. J. MA, Mário ŠPÍREK, K. KANIECKI, Y. KWON, Ondrej BELAN, Kateřina KREJČÍ, S. J. BOULTON, P. SUNG, E. C. GREENE and Lumír KREJČÍ. Single-molecule visualization of human RECQ5 interactions with single-stranded DNA recombination intermediates. \textit{Nucleic acids research}. Oxford: Oxford University Press, 2021, vol.~49, No~1, p.~285-305. ISSN~0305-1048. Available from: https://dx.doi.org/10.1093/nar/gkaa1184.

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