title: The cellular phenotype of Roberts syndrome fibroblasts as revealed by ectopic expression of ESCO2.
abstract: Cohesion between sister chromatids is essential for faithful chromosome segregation. In budding yeast, the acetyltransferase Eco1/Ctf7 establishes cohesion during DNA replication in S phase and in response to DNA double strand breaks in G2/M phase. In humans two Eco1 orthologs exist: ESCO1 and ESCO2. Both proteins are required for proper sister chromatid cohesion, but their exact function is unclear at present. Since ESCO2 has been identified as the gene defective in the rare autosomal recessive cohesinopathy Roberts syndrome (RBS), cells from RBS patients can be used to elucidate the role of ESCO2. We investigated for the first time RBS cells in comparison to isogenic controls that stably express V5- or GFP-tagged ESCO2. We show that the sister chromatid cohesion defect in the transfected cell lines is rescued and suggest that ESCO2 is regulated by proteasomal degradation in a cell cycle-dependent manner. In comparison to the corrected cells RBS cells were hypersensitive to the DNA-damaging agents mitomycin C, camptothecin and etoposide, while no particular sensitivity to UV, ionizing radiation, hydroxyurea or aphidicolin was found. The cohesion defect of RBS cells and their hypersensitivity to DNA-damaging agents were not corrected by a patient-derived ESCO2 acetyltransferase mutant (W539G), indicating that the acetyltransferase activity of ESCO2 is essential for its function. In contrast to a previous study on cells from patients with Cornelia de Lange syndrome, another cohesinopathy, RBS cells failed to exhibit excessive chromosome aberrations after irradiation in G2 phase of the cell cycle. Our results point at an S phase-specific role for ESCO2 in the maintenance of genome stability.
authors.affiliation: Department of Clinical Genetics, VU University Medical Center, Amsterdam, The Netherlands.
authors.forename: Petra
authors.initials: P
authors.lastname: van der Lelij
authors.forename: Barbara C
authors.initials: BC
authors.lastname: Godthelp
authors.forename: Wouter
authors.initials: W
authors.lastname: van Zon
authors.forename: Djoke
authors.initials: D
authors.lastname: van Gosliga
authors.forename: Anneke B
authors.initials: AB
authors.lastname: Oostra
authors.forename: Jûrgen
authors.initials: J
authors.lastname: Steltenpool
authors.forename: Jan
authors.initials: J
authors.lastname: de Groot
authors.forename: Rik J
authors.initials: RJ
authors.lastname: Scheper
authors.forename: Rob M
authors.initials: RM
authors.lastname: Wolthuis
authors.forename: Quinten
authors.initials: Q
authors.lastname: Waisfisz
authors.forename: Firouz
authors.initials: F
authors.lastname: Darroudi
authors.forename: Hans
authors.initials: H
authors.lastname: Joenje
authors.forename: Johan P
authors.initials: JP
authors.lastname: de Winter
mesh_terms: D000123:Acetyltransferases / Q000378:metabolism*; D002166:Camptothecin / Q000494:pharmacology; D018797:Cell Cycle Proteins / Q000378:metabolism; D002868:Chromosomal Proteins, Non-Histone / Q000378:metabolism*; D002869:Chromosome Aberrations; D020090:Chromosome Segregation; D000013:Congenital Abnormalities / Q000175:diagnosis / Q000235:genetics; D004249:DNA Damage; D005047:Etoposide / Q000494:pharmacology; D005347:Fibroblasts / Q000378:metabolism*; D006130:Growth Disorders / Q000175:diagnosis* / Q000235:genetics; D006801:Humans; D007223:Infant; D008297:Male; D016685:Mitomycin / Q000494:pharmacology; D019384:Nucleic Acid Synthesis Inhibitors / Q000494:pharmacology; D012854:Sister Chromatid Exchange; D013577:Syndrome; D000097722:Cohesins
chemical_list: D000123:Acetyltransferases; D002166:Camptothecin; D018797:Cell Cycle Proteins; D002868:Chromosomal Proteins, Non-Histone; D005047:Etoposide; D016685:Mitomycin; D019384:Nucleic Acid Synthesis Inhibitors; D000097722:Cohesins; C500785:ESCO2 protein, human