Evidence that heteronuclear proteins interact with the XIST RNA in vitro

The process of X chromosome inactivation results in the transcriptional silencing of one of the two X chromosomes in mammalian females. A large heterogenous nuclear RNA that is expressed exclusively from the inactive X chromosome (XIST-X Inactive Specific Transcripts) has been implicated in the inactivation process. The XIST RNA colocalizes with the inactive X chromosome and therefore proteins that interact with the XIST RNA may be involved in the inactivation of the X chromosome. In order to identify such proteins we have used an in vitro UV light cross-linking technique to detect nuclear proteins associating with sections of the XIST RNA. The strongest interaction detected by this technique was between a pair of approximately 40 kDa proteins and a 5' region of the XIST RNA which contains a series of well-conserved tandem repeats. Immunoprecipitation suggested that these proteins may be the heteronuclear proteins hnRNPC1/C2.