Final published version
Licence: CC BY: Creative Commons Attribution 4.0 International License
Research output: Contribution to Journal/Magazine › Journal article › peer-review
Research output: Contribution to Journal/Magazine › Journal article › peer-review
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TY - JOUR
T1 - Crucial role of the NSE1 RING domain in Smc5/6 stability and FANCM-independent fork progression
AU - Lorite, Neus P
AU - Apostolova, Sonia
AU - Guasch-Vallés, Marta
AU - Pryer, Aaron
AU - Unzueta, Fernando
AU - Freire, Raimundo
AU - Solé-Soler, Roger
AU - Pedraza, Neus
AU - Dolcet, Xavier
AU - Garí, Eloi
AU - Agell, Neus
AU - Taylor, Elaine M
AU - Colomina, Neus
AU - Torres-Rosell, Jordi
PY - 2024/12/31
Y1 - 2024/12/31
N2 - The Smc5/6 complex is a highly conserved molecular machine involved in the maintenance of genome integrity. While its functions largely depend on restraining the fork remodeling activity of Mph1 in yeast, the presence of an analogous Smc5/6-FANCM regulation in humans remains unknown. We generated human cell lines harboring mutations in the NSE1 subunit of the Smc5/6 complex. Point mutations or truncations in the RING domain of NSE1 result in drastically reduced Smc5/6 protein levels, with differential contribution of the two zinc-coordinating centers in the RING. In addition, nse1-RING mutant cells display cell growth defects, reduced replication fork rates, and increased genomic instability. Notably, our findings uncover a synthetic sick interaction between Smc5/6 and FANCM and show that Smc5/6 controls fork progression and chromosome disjunction in a FANCM-independent manner. Overall, our study demonstrates that the NSE1 RING domain plays vital roles in Smc5/6 complex stability and fork progression through pathways that are not evolutionary conserved.
AB - The Smc5/6 complex is a highly conserved molecular machine involved in the maintenance of genome integrity. While its functions largely depend on restraining the fork remodeling activity of Mph1 in yeast, the presence of an analogous Smc5/6-FANCM regulation in humans remains unknown. We generated human cell lines harboring mutations in the NSE1 subunit of the Smc5/6 complex. Point mutations or truncations in the RING domain of NSE1 result in drastically reduced Smc5/6 protein levels, with differential contribution of the two zinc-coordinating centers in the RING. In addition, nse1-RING mutant cells display cell growth defects, reduced replication fork rates, and increased genomic instability. Notably, our findings uncover a synthetic sick interaction between Smc5/6 and FANCM and show that Smc5/6 controls fork progression and chromosome disjunction in a FANCM-independent manner. Overall, our study demonstrates that the NSE1 RING domain plays vital roles in Smc5/6 complex stability and fork progression through pathways that are not evolutionary conserved.
KW - DNA fiber
KW - NSE4
KW - RING
KW - Fanconi anemia
KW - MMS
KW - Anaphase
KW - SMC6
KW - NSE2
KW - FANCM
KW - NSE3
KW - NSE1
KW - Genomic stability
KW - SMC5
KW - Smc5/6
KW - DNA replication
U2 - 10.1007/s00018-024-05275-3
DO - 10.1007/s00018-024-05275-3
M3 - Journal article
VL - 81
JO - Cellular and Molecular Life Sciences : CMLS
JF - Cellular and Molecular Life Sciences : CMLS
SN - 1420-682X
IS - 1
M1 - 251
ER -