TY - JOUR
T1 - Recapitulation of the Roberts syndrome cellular phenotype by inhibition of INCENP, ZWINT-1 and ZW10 genes
AU - Musio, Antonio
AU - Mariani, Tullio
AU - Montagna, Cristina
AU - Zambroni, Desirèe
AU - Ascoli, Cesare
AU - Ried, Thomas
AU - Vezzoni, Paolo
N1 - Funding Information:
Partially funded by grants from FIRB (RBNE019J9W) and “Progetto Genomica Funzionale” from MIUR and “Progetto Cellule Staminali” from Ministero Salute, Ricerca Finalizzata IRCCS to P.V and FIRB (RBNE01RNN7) to A.M. We thank the Galliera Genetic Bank Italian Telethon project for providing us with Roberts cell line described here. This work is manuscript no. 75 of the Genoma 2000/ITBA Project funded by CARIPLO.
PY - 2004/4/28
Y1 - 2004/4/28
N2 - Roberts syndrome is an autosomal recessive disorder characterised primarily by symmetric reduction of all limbs and growth retardation. Patients have been reported to have premature separation of heterochromatin regions of many chromosomes and abnormalities in cell cycle. Given the rarity of the syndrome, the linkage analysis approach is not suitable to identify the responsible gene. In this work, a cell line derived from a patient affected by Roberts syndrome was characterized by cell biology and molecular cytogenetics, including comparative genomic hybridization and spectral karyotype. No recurrent chromosomal rearrangements were identified. Thereafter, based on the fact that premature chromatide separation is a reliable marker of the disease, we used antisense oligonucleotide technologies to inhibit six genes involved in various steps of the correct chromosome segregation, such as chromosome cohesion, kinetochore assembling, spindle checkpoint and spindle formation. We found that the inhibition of INCENP, ZWINT-1, ZW10 genes results in the appearance of mitotic cells characterised by centromere separation, chromosome aneuploidy and micronuclei formation. In addition, INCENP, ZWINT-1, ZW10 antisense-treated chromosome morphology was very similar to that of Roberts chromosome when analysed by atomic force microscopy. We concluded that INCENP, ZWINT-1, ZW10 gene inhibition results in cellular phenocopies of Roberts syndrome. Taken together, these findings support a possible role of these genes in the pathogenesis of Roberts syndrome.
AB - Roberts syndrome is an autosomal recessive disorder characterised primarily by symmetric reduction of all limbs and growth retardation. Patients have been reported to have premature separation of heterochromatin regions of many chromosomes and abnormalities in cell cycle. Given the rarity of the syndrome, the linkage analysis approach is not suitable to identify the responsible gene. In this work, a cell line derived from a patient affected by Roberts syndrome was characterized by cell biology and molecular cytogenetics, including comparative genomic hybridization and spectral karyotype. No recurrent chromosomal rearrangements were identified. Thereafter, based on the fact that premature chromatide separation is a reliable marker of the disease, we used antisense oligonucleotide technologies to inhibit six genes involved in various steps of the correct chromosome segregation, such as chromosome cohesion, kinetochore assembling, spindle checkpoint and spindle formation. We found that the inhibition of INCENP, ZWINT-1, ZW10 genes results in the appearance of mitotic cells characterised by centromere separation, chromosome aneuploidy and micronuclei formation. In addition, INCENP, ZWINT-1, ZW10 antisense-treated chromosome morphology was very similar to that of Roberts chromosome when analysed by atomic force microscopy. We concluded that INCENP, ZWINT-1, ZW10 gene inhibition results in cellular phenocopies of Roberts syndrome. Taken together, these findings support a possible role of these genes in the pathogenesis of Roberts syndrome.
KW - Cell cycle
KW - Centromere separation
KW - DMEM
KW - Dulbeco's minimal essential medium
KW - FCS
KW - FITC
KW - Flourescein isothiocynate
KW - Foetal calf serum
KW - INCENP
KW - PD
KW - Population doubling
KW - Roberts syndrome
KW - SKY
KW - TUNEL
KW - TdT-medicated dUTP nick end labelling
KW - ZW10
KW - ZWINT-1
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U2 - 10.1016/j.gene.2004.01.028
DO - 10.1016/j.gene.2004.01.028
M3 - Article
C2 - 15094189
AN - SCOPUS:14544308098
SN - 0378-1119
VL - 331
SP - 33
EP - 40
JO - Gene
JF - Gene
IS - 1-2
ER -