Human Molecular Genetics Advance Access published online on January 7, 2009
Human Molecular Genetics, doi:10.1093/hmg/ddp009
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The loss of the snoRNP chaperone Nopp140 from Cajal bodies of patient fibroblasts correlates with the severity of spinal muscular atrophy
1 Laboratoire de Biologie Cellulaire des Membranes, Institut Jacques Monod (IJM), UMR 7592 CNRS/Universités Paris 6 et 7, 2 Place Jussieu, F-75251 Paris cedex 05, France 2 U781 INSERM, IRNEM Institute, Hôpital Necker-Enfants Malades, Paris, France 3 Department of Anatomy and Structural Biology, Albert Einstein College of Medicine, Bronx, NY 10461
* To whom correspondence should be addressed: Dr Suzie Lefebvre, Institut Jacques Monod, UMR 7592 CNRS/Universités Paris 6 et 7, Department of Cell Biology, 2 Place Jussieu, F-75251 Paris cedex 05, France. Tel +33 1 44 27 69 40, Fax +33 1 44 27 59 94, E-mail: lefebvre{at}ijm.jussieu.fr
Received October 16, 2008; Revised January 5, 2008; Accepted January 5, 2008
Spinal Muscular Atrophy (SMA) is a common autosomal recessive neurodegenerative disease caused by reduced SMN levels. The assembly machinery containing SMN is implicated in the biogenesis of the spliceosomal small nuclear ribonucleoproteins, snRNPs. SMN is present in both the cytoplasm and nucleus, where it transiently accumulates in subnuclear domains named Cajal bodies (CBs) and functions in the maturation of snRNPs and small nucleolar (sno)RNPs. The impact of lowering SMN levels on the composition of CBs in SMA cells is still not completely understood. Here, we analyse the CB composition in immortalised and primary fibroblasts from SMA patients. We show that the U snRNA export factors PHAX and CRM1 and the box C/D snoRNP core protein fibrillarin concentrate in CBs from SMA cells, whereas the box H/ACA core proteins GAR1 and NAP57/dyskerin show reduced CB localisation. Remarkably, the functional deficiency in SMA cells is associated with decreased localisation of the snoRNP chaperone Nopp140 in CBs that correlates with disease severity. Indeed, RNA interference knockdown experiments in control fibroblasts demonstrate that SMN is required for the accumulation of Nopp140 in CBs. Conversely, overexpression of SMN in SMA cells restores the CB localisation of Nopp140 whereas SMN mutants found in SMA patients are defective in promoting the association of Nopp140 with CBs. Taken together, we demonstrate that only a subset of CB functions (as indicated by the association of representative factors) are impaired in SMA cells and, importantly, we identify the decrease of Nopp140 localisation in CBs as a phenotypic marker for SMA.