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

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 1752-301X. Available from: https://dx.doi.org/10.3390/cells11010165.

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, 1752-301X.

Other information

• Original language: English
• Type of outcome: Article in a journal
• Field of Study: 10608 Biochemistry and molecular biology
• Country of publisher: Switzerland
• Confidentiality degree: is not subject to a state or trade secret
• WWW: URL
• 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
• Tags: CF PROT, rivok
• Tags: International impact, Reviewed

Abstract

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

• LM2018127, research and development project: Name: Česká infrastruktura pro integrativní strukturní biologii (Acronym: CIISB)

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

• 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