In the female mammal, one of two X-chromosomes is inactivated to ensure that males and females have equal X-chromosome dosage. X-chromosome inactivation (XCI) can take place randomly or in an imprinted manner on the paternal X (XP). In the mouse, random XCI is observed in the embryo proper (soma) and imprinted XCI in the placenta. Both somatic and placental XCI require the 'X-inactivation center'(Xic), an X-linked region known for genes making noncoding RNA (ncRNA). The Xic produces Xist RNA, a large untranslated RNA that 'paints'the X and recruits silencing factors to the X in cis. Our laboratory previously identified the antisense Tsix gene and a second noncoding element, Xite, as essential repressors of Xist. In the previous funding cycle, we focused on the molecular means by which Tsix and Xite regulate different steps of XCI. This investigation has resulted in three major discoveries: (i) In females, the Xs 'pair'transiently with each other at the onset of XCI. We believe that pairing mediates 'cross-talk'between the Xs to ensure mutually exclusive chromosome fates. Tsix and Xite mediate the pairing interaction. (ii) Tsix controls Xist expression through preemptive changes in chromatin structure driven by its antisense transcription. Tsix RNA also binds Dnmt3a and directs de novo DNA methylation to the Xist promoter. (iii) Dosage compensation occurs much earlier in development than previously thought. We believe that imprinted XCI has already taken place by the time of conception and, on this basis, propose that the XP is inherited as a 'pre-inactivated'chromosome from the paternal germline. These discoveries have raised many new questions, which we now intend to pursue in three distinct Aims.
In Aim 1, we will define molecular factors required for X-chromosome pairing.
In Aim 2, we intend to investigate how chromatin modifications are directed by Tsix transcription and what chromatin modifiers are recruited by Tsix RNA.
In Aim 3, we will carry out genetic and biochemical tests to distinguish between the pre-inactivation and de novo models for imprinted XCI. Information gained from the research program will have broad implications for human genetics, cancer, and epigenetic reprogramming. It will also enhance our understanding of basic biological processes, especially those involving chromosome dynamics, ncRNA regulation, and transgenerational inheritance.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM058839-13
Application #
8022963
Study Section
Molecular Genetics C Study Section (MGC)
Program Officer
Carter, Anthony D
Project Start
1999-01-01
Project End
2012-02-19
Budget Start
2011-01-01
Budget End
2012-02-19
Support Year
13
Fiscal Year
2011
Total Cost
$345,584
Indirect Cost
Name
Massachusetts General Hospital
Department
Type
DUNS #
073130411
City
Boston
State
MA
Country
United States
Zip Code
02199
Sun, Sha; Payer, Bernhard; Namekawa, Satoshi et al. (2015) Xist imprinting is promoted by the hemizygous (unpaired) state in the male germ line. Proc Natl Acad Sci U S A 112:14415-22
Kung, Johnny T; Kesner, Barry; An, Jee Young et al. (2015) Locus-specific targeting to the X chromosome revealed by the RNA interactome of CTCF. Mol Cell 57:361-75
Yang, Lin; Froberg, John E; Lee, Jeannie T (2014) Long noncoding RNAs: fresh perspectives into the RNA world. Trends Biochem Sci 39:35-43
Lee, Jeannie T; Bartolomei, Marisa S (2013) X-inactivation, imprinting, and long noncoding RNAs in health and disease. Cell 152:1308-23
Sun, Sha; Del Rosario, Brian C; Szanto, Attila et al. (2013) Jpx RNA activates Xist by evicting CTCF. Cell 153:1537-51
Payer, Bernhard; Rosenberg, Michael; Yamaji, Masashi et al. (2013) Tsix RNA and the germline factor, PRDM14, link X reactivation and stem cell reprogramming. Mol Cell 52:805-18
Kung, Johnny T Y; Colognori, David; Lee, Jeannie T (2013) Long noncoding RNAs: past, present, and future. Genetics 193:651-69
Lee, Jeannie T (2012) Epigenetic regulation by long noncoding RNAs. Science 338:1435-9
Erwin, Jennifer A; del Rosario, Brian; Payer, Bernhard et al. (2012) An ex vivo model for imprinting: mutually exclusive binding of Cdx2 and Oct4 as a switch for imprinted and random X-inactivation. Genetics 192:857-68
Anguera, Montserrat C; Sadreyev, Ruslan; Zhang, Zhaoqing et al. (2012) Molecular signatures of human induced pluripotent stem cells highlight sex differences and cancer genes. Cell Stem Cell 11:75-90

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