The Ewing sarcoma protein EWS belongs to the FET family (FUS, EWS, TAF15) of polypeptides, which can bind RNA as well as DNA and are implicated in transcription, splicing, RNA transport, signalling and maintenance of genomic integrity. EWS interacts with U1C, SF1 and several SR proteins and hnRNPs. To identify potential alternative splicing events regulated by EWS, RNA from cells depleted of EWS or the corresponding controls were hybridized to a splicing sensitive microarray developed in our laboratory, which analyzes 1804 events in 482 genes relevant for cancer progression and RNA processing. Using this platform, we have identified 39 alternative splicing events in 31 genes affected by EWS knockdown. These changes are enriched in alternative acceptor choices and depleted in exon skipping events. Interestingly, alternative splicing changes were identified that affect key genes involved in the response to DNA damage. Remarkably, some of these changes were also induced upon UV irradiation of control cells. This observation suggests the possibility that changes in EWS activity are part of the mechanisms underlying the changes in alternative splicing induced by genotoxic stress. Consistent with this possibility, we have observed a remarkable change in subcellular localization of the EWS protein upon UV irradiation. These results also offer a potential explanation for the observation that EWS knockout mice show hypersensitivity to ionizing radiation and premature senescence.
Regulation of alternative splicing by the Ewing Sarcoma Protein (EWS) and DNA damage
Paronetto MP;
2010-01-01
Abstract
The Ewing sarcoma protein EWS belongs to the FET family (FUS, EWS, TAF15) of polypeptides, which can bind RNA as well as DNA and are implicated in transcription, splicing, RNA transport, signalling and maintenance of genomic integrity. EWS interacts with U1C, SF1 and several SR proteins and hnRNPs. To identify potential alternative splicing events regulated by EWS, RNA from cells depleted of EWS or the corresponding controls were hybridized to a splicing sensitive microarray developed in our laboratory, which analyzes 1804 events in 482 genes relevant for cancer progression and RNA processing. Using this platform, we have identified 39 alternative splicing events in 31 genes affected by EWS knockdown. These changes are enriched in alternative acceptor choices and depleted in exon skipping events. Interestingly, alternative splicing changes were identified that affect key genes involved in the response to DNA damage. Remarkably, some of these changes were also induced upon UV irradiation of control cells. This observation suggests the possibility that changes in EWS activity are part of the mechanisms underlying the changes in alternative splicing induced by genotoxic stress. Consistent with this possibility, we have observed a remarkable change in subcellular localization of the EWS protein upon UV irradiation. These results also offer a potential explanation for the observation that EWS knockout mice show hypersensitivity to ionizing radiation and premature senescence.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.