Tsix, a gene antisense to Xist at the X-inactivation centre

Abstract

In mammals, dosage compensation is achieved by X inactivation1 and is regulated in cis by the X-inactivation centre2 (Xic) and Xist (refs 3,4,5). The Xic controls X-chromosome counting, choice of X to inactivate and initiation of silencing. Xic action culminates in a change in Xist RNA property from a scarce, unstable RNA (Refs 6,7) to highly expressed Xist RNA that coats the future inactive X (ref. 8). Deleting a 65-kb region downstream of Xist results in constitutive Xist expression and X inactivation, implying the presence of a cis-regulatory element9. In this region, we now report the discovery of a gene antisense to Xist. Tsix is a 40-kb RNA originating 15 kb downstream of Xist and transcribed across the Xist locus. Tsix sequence is conserved at the human XIC. Tsix RNA has no conserved ORFs, is seen exclusively in the nucleus and is localized at Xic. Before the onset of X inactivation, Tsix is expressed from both X chromosomes. At the onset of X inactivation, Tsix expression becomes monoallelic, is associated with the future active X and persists until Xist is turned off. Tsix is not found on the inactive X once cells enter the X-inactivation pathway. Tsix has features suggesting a role in regulating the early steps of X inactivation, but not the silencing step.

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Figure 1: Strand-specific FISH probes reveal an antisense RNA crossing the Xist locus.
Figure 2: Mapping Tsix by RNA FISH.
Figure 3: Tsix begins 15 kb downstream of Xist and spans 40 kb of sequence.
Figure 4: Tsix is dynamically regulated during female and male EB differentiation.
Figure 5: Tsix RNA co-localizes with Xist RNA and is exclusively nuclear.
Figure 6: The mouse Tsix sequence is conserved at the human XIC.

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Acknowledgements

We thank E. Li for the generous gift of EL16; J.E. Kirby, B. Kingston, M. Oettinger, N. Stavropoulos and Y. Ogawa for critique of the manuscript. This work was supported by an American Cancer Society Grant (ACS-IRG 173-H) and a Hoechst AG award to J.T.L.

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Correspondence to Jeannie Lee.

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