The EWS/NOR1 fusion gene product gains a novel activity affecting pre-mRNA splicing.
In extraskeletal myxoid chondrosarcoma, chromosomal translocation creates a gene fusion between EWS and the orphan nuclear receptor NOR1. The resulting fusion gene product, EWS/NOR1, has been believed to lead to malignant transformation by functioning as a transcriptional activator, but an alternative mechanism may also be involved. Here, using a newly developed functional complementation screening in yeast, we found that EWS/NOR1, but not EWS or NOR1, complemented the loss of function of the small nuclear ribonucleoprotein Snu23p, an essential factor for pre-mRNA splicing in yeast. To verify the potential function of EWS/NOR1 in mammalian cells, we next showed that overexpression of EWS/NOR1 caused increased usage of the distal 5'-splice site of pre-mRNA splicing and that EWS/NOR1 interacted with the human splicing protein U1C; neither EWS nor NOR1 had the same activity or interaction as EWS/NOR1. Altogether, our findings reveal that EWS/NOR1 gains a novel activity affecting pre-mRNA splicing.
Ohkura
Naganari
N
Growth Factor Division, National Cancer Center Research Institute, 5-1-1 Tsukiji, Chuo-ku, Tokyo 104-0045, Japan. nohkura@gan2.ncc.go.jp
Yaguchi
Hiroko
H
Tsukada
Toshihiko
T
Yamaguchi
Ken
K
D017500:Adenovirus E1A Proteins / Q000235:genetics; D000595:Amino Acid Sequence; D000818:Animals; D002467:Cell Nucleus / Q000737:chemistry; D004268:DNA-Binding Proteins / Q000032:analysis / Q000502:physiology*; D034441:Heterogeneous-Nuclear Ribonucleoproteins; D006801:Humans; D008969:Molecular Sequence Data; D009419:Nerve Tissue Proteins / Q000032:analysis / Q000502:physiology*; D012322:RNA Precursors / Q000235:genetics*; D012326:RNA Splicing; D034802:RNA-Binding Protein EWS; D011987:Receptors, Steroid; D011988:Receptors, Thyroid Hormone; D011993:Recombinant Fusion Proteins / Q000032:analysis / Q000502:physiology*; D012261:Ribonucleoproteins / Q000032:analysis / Q000502:physiology*; D017411:Ribonucleoproteins, Small Nuclear / Q000378:metabolism
D017500:Adenovirus E1A Proteins; D004268:DNA-Binding Proteins; D034441:Heterogeneous-Nuclear Ribonucleoproteins; C490412:NR4A3 protein, human; D009419:Nerve Tissue Proteins; C490411:Nr4a3 protein, mouse; D012322:RNA Precursors; D034802:RNA-Binding Protein EWS; D011987:Receptors, Steroid; D011988:Receptors, Thyroid Hormone; D011993:Recombinant Fusion Proteins; D012261:Ribonucleoproteins; D017411:Ribonucleoproteins, Small Nuclear; C491260:SNRPC protein, human