The Phenotype of Physcomitrium patens SMC6 Mutant with
Interrupted Hinge Interactions

J 2025

The Phenotype of Physcomitrium patens SMC6 Mutant with Interrupted Hinge Interactions

ANGELIS, Karel J.; Marcela HOLA; Radka VAGNEROVA; Jitka VACULÍKOVÁ; Jan PALECEK et al.

Základní údaje

Originální název

The Phenotype of Physcomitrium patens SMC6 Mutant with Interrupted Hinge Interactions

Autoři

ANGELIS, Karel J.; Marcela HOLA; Radka VAGNEROVA; Jitka VACULÍKOVÁ a Jan PALECEK

Vydání

Genes, BASEL, MDPI, 2025, 2073-4425

Další údaje

Jazyk

angličtina

Typ výsledku

Článek v odborném periodiku

Obor

10608 Biochemistry and molecular biology

Stát vydavatele

Švýcarsko

Utajení

není předmětem státního či obchodního tajemství

Odkazy

URL

Impakt faktor

Impact factor: 2.800 v roce 2024

Označené pro přenos do RIV

Ano

Organizační jednotka

Přírodovědecká fakulta

Klíčová slova anglicky

SMC5/6; hinge domain; protein-protein interactions; DSB repair; mutagenesis; gene targeting; rDNA stability; protonemata development; Physcomitrium patens SMC5/6

Štítky

143160, rivok

Příznaky

Mezinárodní význam, Recenzováno

Změněno: 14. 1. 2026 14:16, Mgr. Marie Novosadová Šípková, DiS.

Anotace

V originále

Background/Objectives: The Structural Maintenance of Chromosomes (SMC) proteins form essential heterocomplexes for the preservation of DNA structure and its functions, and hence cell viability. The SMC5/6 dimer is assembled by direct interactions of ATP heads via the kleisin NSE4 bridge and by SMC hinges. The structure might be interrupted by a single point mutation within a conserved motif of the SMC6-hinge. We describe the phenomena associated with the impairment of the SMC5/6 complex with morphology, repair of DNA double strand breaks (DSB), mutagenesis, recombination and gene targeting (GT) in the moss Physcomitrium patens (P. patens). Methods: Using CRISPR/Cas9-directed oligonucleotide replacement, we have introduced two close G to R point mutations in the hinge domain of SMC6 of P. patens and show that both mutations are not toxic and allow viability of mutant lines. Results: The G514R mutation fully prevents the interaction of SMC6 not only with SMC5, but also with NSE5 and NSE6, while the mutation at G517R has no effect. The Ppsmc6_G514R line has aberrant morphology, spontaneous and bleomycin-induced mutagenesis, and maintenance of the number of rDNA copies. The most unique feature is the interference with gene targeting (GT), which is completely abolished. In contrast, the Ppsmc6_G517R line is close to WT in many aspects. Surprisingly, both mutations have no direct effect on the rate of DSB repair in dividing and differentiated cells. Conclusions: Abolished interactions of SMC6 with SMC5 and NSE5,6 partners, which allow DSB repair, but impair other repair and recombination functions, suggests also regulatory role for SMC6.

Návaznosti

GA23-05284S, projekt VaV

Název: Rozluštění vztahu mezi homologní rekombinací a umlčením genů

Investor: Grantová agentura ČR, Rozluštění vztahu mezi homologní rekombinací a umlčením genů

Citovat

ANGELIS, Karel J.; Marcela HOLA; Radka VAGNEROVA; Jitka VACULÍKOVÁ a Jan PALECEK. The Phenotype of Physcomitrium patens SMC6 Mutant with Interrupted Hinge Interactions. Genes. BASEL: MDPI, 2025, roč. 16, č. 9, s. 1-15. ISSN 2073-4425. Dostupné z: https://doi.org/10.3390/genes16091091.

@article{2547062,
   author = {Angelis, Karel J. and Hola, Marcela and Vagnerova, Radka and Vaculíková, Jitka and Palecek, Jan},
   article_location = {BASEL},
   article_number = {9},
   doi = {https://doi.org/10.3390/genes16091091},
   keywords = {SMC5/6; hinge domain; protein-protein interactions; DSB repair; mutagenesis; gene targeting; rDNA stability; protonemata development; Physcomitrium patens SMC5/6},
   language = {eng},
   issn = {2073-4425},
   journal = {Genes},
   title = {The Phenotype of Physcomitrium patens SMC6 Mutant with Interrupted Hinge Interactions},
   url = {https://www.mdpi.com/2073-4425/16/9/1091},
   volume = {16},
   year = {2025}
}

TY  - JOUR
ID  - 2547062
AU  - Angelis, Karel J. - Hola, Marcela - Vagnerova, Radka - Vaculíková, Jitka - Palecek, Jan
PY  - 2025
TI  - The Phenotype of Physcomitrium patens SMC6 Mutant with Interrupted Hinge Interactions
JF  - Genes
VL  - 16
IS  - 9
SP  - 1-15
EP  - 1-15
PB  - MDPI
SN  - 20734425
KW  - SMC5/6
KW  - hinge domain
KW  - protein-protein interactions
KW  - DSB repair
KW  - mutagenesis
KW  - gene targeting
KW  - rDNA stability
KW  - protonemata development
KW  - Physcomitrium patens SMC5/6
UR  - https://www.mdpi.com/2073-4425/16/9/1091
N2  - Background/Objectives: The Structural Maintenance of Chromosomes (SMC) proteins form essential heterocomplexes for the preservation of DNA structure and its functions, and hence cell viability. The SMC5/6 dimer is assembled by direct interactions of ATP heads via the kleisin NSE4 bridge and by SMC hinges. The structure might be interrupted by a single point mutation within a conserved motif of the SMC6-hinge. We describe the phenomena associated with the impairment of the SMC5/6 complex with morphology, repair of DNA double strand breaks (DSB), mutagenesis, recombination and gene targeting (GT) in the moss Physcomitrium patens (P. patens). Methods: Using CRISPR/Cas9-directed oligonucleotide replacement, we have introduced two close G to R point mutations in the hinge domain of SMC6 of P. patens and show that both mutations are not toxic and allow viability of mutant lines. Results: The G514R mutation fully prevents the interaction of SMC6 not only with SMC5, but also with NSE5 and NSE6, while the mutation at G517R has no effect. The Ppsmc6_G514R line has aberrant morphology, spontaneous and bleomycin-induced mutagenesis, and maintenance of the number of rDNA copies. The most unique feature is the interference with gene targeting (GT), which is completely abolished. In contrast, the Ppsmc6_G517R line is close to WT in many aspects. Surprisingly, both mutations have no direct effect on the rate of DSB repair in dividing and differentiated cells. Conclusions: Abolished interactions of SMC6 with SMC5 and NSE5,6 partners, which allow DSB repair, but impair other repair and recombination functions, suggests also regulatory role for SMC6.
ER  -

ANGELIS, Karel J.; Marcela HOLA; Radka VAGNEROVA; Jitka VACULÍKOVÁ a Jan PALECEK. The Phenotype of Physcomitrium patens SMC6 Mutant with Interrupted Hinge Interactions. \textit{Genes}. BASEL: MDPI, 2025, roč.~16, č.~9, s.~1-15. ISSN~2073-4425. Dostupné z: https://doi.org/10.3390/genes16091091.

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