Role of Nse1 Subunit of SMC5/6 Complex as a Ubiquitin Ligase

J 2022

Role of Nse1 Subunit of SMC5/6 Complex as a Ubiquitin Ligase

KOLESÁR, Peter, Karel STEJSKAL, David POTĚŠIL, Johanne M MURRAY, Jan PALEČEK et. al.

Basic information

Original name

Role of Nse1 Subunit of SMC5/6 Complex as a Ubiquitin Ligase

Name in Czech

Úloha Nse1 podjednotky komplexu SMC5/6 v ubikvitinaci

Authors

KOLESÁR, Peter (703 Slovakia, belonging to the institution), Karel STEJSKAL (203 Czech Republic, belonging to the institution), David POTĚŠIL (203 Czech Republic, belonging to the institution), Johanne M MURRAY and Jan PALEČEK (203 Czech Republic, guarantor, belonging to the institution)

Edition

Cells, BASEL, MDPI, 2022, 2073-4409

Other information

Language

English

Type of outcome

Článek v odborném periodiku

Field of Study

10608 Biochemistry and molecular biology

Country of publisher

Switzerland

Confidentiality degree

není předmětem státního či obchodního tajemství

References:

URL

Impact factor

Impact factor: 6.000

RIV identification code

RIV/00216224:14310/22:00126432

Organization unit

Faculty of Science

DOI

http://dx.doi.org/10.3390/cells11010165

UT WoS

000758383300001

Keywords in English

SMC5; 6; Nse1; ubiquitin ligase; ubiquitination; Ubc13; Mms2; Nse4 kleisin

Abstract

V originále

Structural Maintenance of Chromosomes (SMC) complexes are important for many aspects of the chromosomal organization. Unlike cohesin and condensin, the SMC5/6 complex contains a variant RING domain carried by its Nse1 subunit. RING domains are characteristic for ubiquitin ligases, and human NSE1 has been shown to possess ubiquitin-ligase activity in vitro. However, other studies were unable to show such activity. Here, we confirm Nse1 ubiquitin-ligase activity using purified Schizosaccharomyces pombe proteins. We demonstrate that the Nse1 ligase activity is stimulated by Nse3 and Nse4. We show that Nse1 specifically utilizes Ubc13/Mms2 E2 enzyme and interacts directly with ubiquitin. We identify the Nse1 mutation (R188E) that specifically disrupts its E3 activity and demonstrate that the Nse1-dependent ubiquitination is particularly important under replication stress. Moreover, we determine Nse4 (lysine K181) as the first known SMC5/6-associated Nse1 substrate. Interestingly, abolition of Nse4 modification at K181 leads to suppression of DNA-damage sensitivity of other SMC5/6 mutants. Altogether, this study brings new evidence for Nse1 ubiquitin ligase activity, significantly advancing our understanding of this enigmatic SMC5/6 function.

Links

GA18-02067S, research and development project

Name: Úloha Nse1/Nse3/Nse4, E3-ubikvitin ligásy, ve stabilitě genomu

Investor: Czech Science Foundation

LQ1601, research and development project

Name: CEITEC 2020 (Acronym: CEITEC2020)

Investor: Ministry of Education, Youth and Sports of the CR

LTC20033, research and development project

Name: The SMC5/6 complex in nucleome organization (Acronym: Nukleom)

Investor: Ministry of Education, Youth and Sports of the CR, INTER-COST

90127, large research infrastructures

Name: CIISB II

Citovat

KOLESÁR, Peter, Karel STEJSKAL, David POTĚŠIL, Johanne M MURRAY and Jan PALEČEK. Role of Nse1 Subunit of SMC5/6 Complex as a Ubiquitin Ligase. Cells. BASEL: MDPI, 2022, vol. 11, No 1, p. 1-13. ISSN 2073-4409. Available from: https://dx.doi.org/10.3390/cells11010165.

@article{2211224,
   author = {Kolesár, Peter and Stejskal, Karel and Potěšil, David and Murray, Johanne M and Paleček, Jan},
   article_location = {BASEL},
   article_number = {1},
   doi = {http://dx.doi.org/10.3390/cells11010165},
   keywords = {SMC5; 6; Nse1; ubiquitin ligase; ubiquitination; Ubc13; Mms2; Nse4 kleisin},
   language = {eng},
   issn = {2073-4409},
   journal = {Cells},
   title = {Role of Nse1 Subunit of SMC5/6 Complex as a Ubiquitin Ligase},
   url = {https://www.mdpi.com/2073-4409/11/1/165},
   volume = {11},
   year = {2022}
}

TY  - JOUR
ID  - 2211224
AU  - Kolesár, Peter - Stejskal, Karel - Potěšil, David - Murray, Johanne M - Paleček, Jan
PY  - 2022
TI  - Role of Nse1 Subunit of SMC5/6 Complex as a Ubiquitin Ligase
JF  - Cells
VL  - 11
IS  - 1
SP  - 1-13
EP  - 1-13
PB  - MDPI
SN  - 20734409
KW  - SMC5
KW  - 6
KW  - Nse1
KW  - ubiquitin ligase
KW  - ubiquitination
KW  - Ubc13
KW  - Mms2
KW  - Nse4 kleisin
UR  - https://www.mdpi.com/2073-4409/11/1/165
N2  - Structural Maintenance of Chromosomes (SMC) complexes are important for many aspects of the chromosomal organization. Unlike cohesin and condensin, the SMC5/6 complex contains a variant RING domain carried by its Nse1 subunit. RING domains are characteristic for ubiquitin ligases, and human NSE1 has been shown to possess ubiquitin-ligase activity in vitro. However, other studies were unable to show such activity. Here, we confirm Nse1 ubiquitin-ligase activity using purified Schizosaccharomyces pombe proteins. We demonstrate that the Nse1 ligase activity is stimulated by Nse3 and Nse4. We show that Nse1 specifically utilizes Ubc13/Mms2 E2 enzyme and interacts directly with ubiquitin. We identify the Nse1 mutation (R188E) that specifically disrupts its E3 activity and demonstrate that the Nse1-dependent ubiquitination is particularly important under replication stress. Moreover, we determine Nse4 (lysine K181) as the first known SMC5/6-associated Nse1 substrate. Interestingly, abolition of Nse4 modification at K181 leads to suppression of DNA-damage sensitivity of other SMC5/6 mutants. Altogether, this study brings new evidence for Nse1 ubiquitin ligase activity, significantly advancing our understanding of this enigmatic SMC5/6 function.
ER  -

KOLESÁR, Peter, Karel STEJSKAL, David POTĚŠIL, Johanne M MURRAY and Jan PALEČEK. Role of Nse1 Subunit of SMC5/6 Complex as a Ubiquitin Ligase. \textit{Cells}. BASEL: MDPI, 2022, vol.~11, No~1, p.~1-13. ISSN~2073-4409. Available from: https://dx.doi.org/10.3390/cells11010165.

Detailed Information on Publication Record