Sumoylation of the Rad1 nuclease promotes DNA repair and
regulates its DNA association

J 2014

Sumoylation of the Rad1 nuclease promotes DNA repair and regulates its DNA association

SARANGI, Prabha, Zdenka BARTOŠOVÁ, Veronika ALTMANNOVÁ, Cory HOLLAND, Melita CHAVDAROVA et. al.

Basic information

Original name

Sumoylation of the Rad1 nuclease promotes DNA repair and regulates its DNA association

Authors

SARANGI, Prabha (840 United States of America), Zdenka BARTOŠOVÁ (703 Slovakia, belonging to the institution), Veronika ALTMANNOVÁ (203 Czech Republic, belonging to the institution), Cory HOLLAND (840 United States of America), Melita CHAVDAROVA (807 North Macedonia, belonging to the institution), Sang Eun LEE (840 United States of America), Lumír KREJČÍ (203 Czech Republic, guarantor, belonging to the institution) and Xiaolan ZHAO (840 United States of America)

Edition

Nucleic Acids Research, Oxford University Press, 2014, 0305-1048

Other information

Language

English

Type of outcome

Článek v odborném periodiku

Field of Study

10600 1.6 Biological sciences

Country of publisher

United Kingdom of Great Britain and Northern Ireland

Confidentiality degree

není předmětem státního či obchodního tajemství

Impact factor

Impact factor: 9.112

RIV identification code

RIV/00216224:14110/14:00073652

Organization unit

Faculty of Medicine

DOI

http://dx.doi.org/10.1093/nar/gku300

UT WoS

000338768100030

Keywords in English

NUCLEOTIDE EXCISION-REPAIR; TRANSCRIPTION FACTOR TFIIH; SACCHAROMYCES-CEREVISIAE; IN-VIVO; HOMOLOGOUS RECOMBINATION; BUDDING YEAST; RAD1-RAD10 NUCLEASE; DAMAGE RECOGNITION; SUMO MODIFICATION; STRAND BREAKS

Abstract

V originále

The Saccharomyces cerevisiae Rad1-Rad10 complex is a conserved, structure-specific endonuclease important for repairing multiple types of DNA lesions. Upon recruitment to lesion sites, Rad1-Rad10 removes damaged sequences, enabling subsequent gap filling and ligation. Acting at mid-steps of repair, the association and dissociation of Rad1-Rad10 with DNA can influence repair efficiency. We show that genotoxin-enhanced Rad1 sumoylation occurs after the nuclease is recruited to lesion sites. A single lysine outside Rad1's nuclease and Rad10-binding domains is sumoylated in vivo and in vitro. Mutation of this site to arginine abolishes Rad1 sumoylation and impairs Rad1-mediated repair at high doses of DNA damage, but sustains the repair of a single double-stranded break. The timing of Rad1 sumoylation and the phenotype bias toward high lesion loads point to a post-incision role for sumoylation, possibly affecting Rad1 dissociation from DNA. Indeed, biochemical examination shows that sumoylation of Rad1 decreases the complex's affinity for DNA without affecting other protein properties. These findings suggest a model whereby sumoylation of Rad1 promotes its disengagement from DNA after nuclease cleavage, allowing it to efficiently attend to large numbers of DNA lesions.

Links

EE2.3.30.0009, research and development project

Name: Zaměstnáním čerstvých absolventů doktorského studia k vědecké excelenci

GAP207/12/2323, research and development project

Name: Endonuleazová a translokázová aktivita v restričních-modifikáčních komplexéch typu I

Investor: Czech Science Foundation

GA13-26629S, research and development project

Name: SUMO a stability genomu

Investor: Czech Science Foundation

Citovat

SARANGI, Prabha, Zdenka BARTOŠOVÁ, Veronika ALTMANNOVÁ, Cory HOLLAND, Melita CHAVDAROVA, Sang Eun LEE, Lumír KREJČÍ and Xiaolan ZHAO. Sumoylation of the Rad1 nuclease promotes DNA repair and regulates its DNA association. Nucleic Acids Research. Oxford University Press, 2014, vol. 42, No 10, p. 6393-6404. ISSN 0305-1048. Available from: https://dx.doi.org/10.1093/nar/gku300.

@article{1184062,
   author = {Sarangi, Prabha and Bartošová, Zdenka and Altmannová, Veronika and Holland, Cory and Chavdarova, Melita and Lee, Sang Eun and Krejčí, Lumír and Zhao, Xiaolan},
   article_number = {10},
   doi = {http://dx.doi.org/10.1093/nar/gku300},
   keywords = {NUCLEOTIDE EXCISION-REPAIR; TRANSCRIPTION FACTOR TFIIH; SACCHAROMYCES-CEREVISIAE; IN-VIVO; HOMOLOGOUS RECOMBINATION; BUDDING YEAST; RAD1-RAD10 NUCLEASE; DAMAGE RECOGNITION; SUMO MODIFICATION; STRAND BREAKS},
   language = {eng},
   issn = {0305-1048},
   journal = {Nucleic Acids Research},
   title = {Sumoylation of the Rad1 nuclease promotes DNA repair and regulates its DNA association},
   volume = {42},
   year = {2014}
}

TY  - JOUR
ID  - 1184062
AU  - Sarangi, Prabha - Bartošová, Zdenka - Altmannová, Veronika - Holland, Cory - Chavdarova, Melita - Lee, Sang Eun - Krejčí, Lumír - Zhao, Xiaolan
PY  - 2014
TI  - Sumoylation of the Rad1 nuclease promotes DNA repair and regulates its DNA association
JF  - Nucleic Acids Research
VL  - 42
IS  - 10
SP  - 6393-6404
EP  - 6393-6404
PB  - Oxford University Press
SN  - 03051048
KW  - NUCLEOTIDE EXCISION-REPAIR
KW  - TRANSCRIPTION FACTOR TFIIH
KW  - SACCHAROMYCES-CEREVISIAE
KW  - IN-VIVO
KW  - HOMOLOGOUS RECOMBINATION
KW  - BUDDING YEAST
KW  - RAD1-RAD10 NUCLEASE
KW  - DAMAGE RECOGNITION
KW  - SUMO MODIFICATION
KW  - STRAND BREAKS
N2  - The Saccharomyces cerevisiae Rad1-Rad10 complex is a conserved, structure-specific endonuclease important for repairing multiple types of DNA lesions. Upon recruitment to lesion sites, Rad1-Rad10 removes damaged sequences, enabling subsequent gap filling and ligation. Acting at mid-steps of repair, the association and dissociation of Rad1-Rad10 with DNA can influence repair efficiency. We show that genotoxin-enhanced Rad1 sumoylation occurs after the nuclease is recruited to lesion sites. A single lysine outside Rad1's nuclease and Rad10-binding domains is sumoylated in vivo and in vitro. Mutation of this site to arginine abolishes Rad1 sumoylation and impairs Rad1-mediated repair at high doses of DNA damage, but sustains the repair of a single double-stranded break. The timing of Rad1 sumoylation and the phenotype bias toward high lesion loads point to a post-incision role for sumoylation, possibly affecting Rad1 dissociation from DNA. Indeed, biochemical examination shows that sumoylation of Rad1 decreases the complex's affinity for DNA without affecting other protein properties. These findings suggest a model whereby sumoylation of Rad1 promotes its disengagement from DNA after nuclease cleavage, allowing it to efficiently attend to large numbers of DNA lesions.
ER  -

SARANGI, Prabha, Zdenka BARTOŠOVÁ, Veronika ALTMANNOVÁ, Cory HOLLAND, Melita CHAVDAROVA, Sang Eun LEE, Lumír KREJČÍ and Xiaolan ZHAO. Sumoylation of the Rad1 nuclease promotes DNA repair and regulates its DNA association. \textit{Nucleic Acids Research}. Oxford University Press, 2014, vol.~42, No~10, p.~6393-6404. ISSN~0305-1048. Available from: https://dx.doi.org/10.1093/nar/gku300.

Detailed Information on Publication Record