The DNA Replication Program Is Altered at the FMR1 Locus in Fragile X Embryonic Stem Cells

Jeannine Gerhardt; Mark J. Tomishima; Nikica Zaninovic; Dilek Colak; Zi Yan; Qiansheng Zhan; Zev Rosenwaks; Samie R. Jaffrey; Carl L. Schildkraut

doi:10.1016/j.molcel.2013.10.029

The DNA Replication Program Is Altered at the FMR1 Locus in Fragile X Embryonic Stem Cells

Jeannine Gerhardt, Mark J. Tomishima, Nikica Zaninovic, Dilek Colak, Zi Yan, Qiansheng Zhan, Zev Rosenwaks, Samie R. Jaffrey, Carl L. Schildkraut

Cell Biology

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83 Scopus citations

Abstract

Fragile X syndrome (FXS) is caused by a CGG repeat expansion in the FMR1 gene that appears to occur during oogenesis and during early embryogenesis. One model proposes that repeat instability depends on the replication fork direction through the repeats such that (CNG)_n hairpin-like structures form, causing DNA polymerase to stall and slip. Examining DNA replication fork progression on single DNA molecules at the endogenous FMR1 locus revealed that replication forks stall at CGG repeats in human cells. Furthermore, replication profiles of FXS human embryonic stem cells (hESCs) compared to nonaffected hESCs showed that fork direction through the repeats is altered at the FMR1 locus in FXS hESCs, such that predominantly the CCG strand serves as the lagging-strand template. This is due to the absence of replication initiation that would typically occur upstream of FMR1, suggesting that altered replication origin usage combined with fork stalling promotes repeat instability during early embryonic development.

Original language	English (US)
Pages (from-to)	19-31
Number of pages	13
Journal	Molecular Cell
Volume	53
Issue number	1
DOIs	https://doi.org/10.1016/j.molcel.2013.10.029
State	Published - Jan 9 2014

ASJC Scopus subject areas

Molecular Biology
Cell Biology

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10.1016/j.molcel.2013.10.029

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@article{f75da8e836414cab841b78f4f1be6848,

title = "The DNA Replication Program Is Altered at the FMR1 Locus in Fragile X Embryonic Stem Cells",

abstract = "Fragile X syndrome (FXS) is caused by a CGG repeat expansion in the FMR1 gene that appears to occur during oogenesis and during early embryogenesis. One model proposes that repeat instability depends on the replication fork direction through the repeats such that (CNG)n hairpin-like structures form, causing DNA polymerase to stall and slip. Examining DNA replication fork progression on single DNA molecules at the endogenous FMR1 locus revealed that replication forks stall at CGG repeats in human cells. Furthermore, replication profiles of FXS human embryonic stem cells (hESCs) compared to nonaffected hESCs showed that fork direction through the repeats is altered at the FMR1 locus in FXS hESCs, such that predominantly the CCG strand serves as the lagging-strand template. This is due to the absence of replication initiation that would typically occur upstream of FMR1, suggesting that altered replication origin usage combined with fork stalling promotes repeat instability during early embryonic development.",

author = "Jeannine Gerhardt and Tomishima, {Mark J.} and Nikica Zaninovic and Dilek Colak and Zi Yan and Qiansheng Zhan and Zev Rosenwaks and Jaffrey, {Samie R.} and Schildkraut, {Carl L.}",

note = "Funding Information: We thank Sally Nolin, Carl Dobkin, Arne Gennerich, Matthew Nicholas, Anne Glicksman, Nicole Ersalesi, Paolo Norio, ChanJung Chang, Solen Gokhan, Sherri Schultz, Andrew Levitas, William C. Drosopoulos, and Settapong T. Kosiyatrakul for valuable experimental suggestions and discussion of the results. This work was supported by NIH/NIGMS 5R01-GM045751 (C.L.S.) and the Empire State Stem Cell Fund through New York State contracts C024348 (C.L.S.) and C024175 (M.J.T), the Starr Tri-Institutional Stem Cell Initiative (M.J.T., Z.R., N.Z., and S.R.J.), and NIH/NIMH 5R01MH80420 (S.R.J.). ",

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doi = "10.1016/j.molcel.2013.10.029",

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T1 - The DNA Replication Program Is Altered at the FMR1 Locus in Fragile X Embryonic Stem Cells

AU - Gerhardt, Jeannine

AU - Tomishima, Mark J.

AU - Zaninovic, Nikica

AU - Colak, Dilek

AU - Yan, Zi

AU - Zhan, Qiansheng

AU - Rosenwaks, Zev

AU - Jaffrey, Samie R.

AU - Schildkraut, Carl L.

N1 - Funding Information: We thank Sally Nolin, Carl Dobkin, Arne Gennerich, Matthew Nicholas, Anne Glicksman, Nicole Ersalesi, Paolo Norio, ChanJung Chang, Solen Gokhan, Sherri Schultz, Andrew Levitas, William C. Drosopoulos, and Settapong T. Kosiyatrakul for valuable experimental suggestions and discussion of the results. This work was supported by NIH/NIGMS 5R01-GM045751 (C.L.S.) and the Empire State Stem Cell Fund through New York State contracts C024348 (C.L.S.) and C024175 (M.J.T), the Starr Tri-Institutional Stem Cell Initiative (M.J.T., Z.R., N.Z., and S.R.J.), and NIH/NIMH 5R01MH80420 (S.R.J.).

PY - 2014/1/9

Y1 - 2014/1/9

N2 - Fragile X syndrome (FXS) is caused by a CGG repeat expansion in the FMR1 gene that appears to occur during oogenesis and during early embryogenesis. One model proposes that repeat instability depends on the replication fork direction through the repeats such that (CNG)n hairpin-like structures form, causing DNA polymerase to stall and slip. Examining DNA replication fork progression on single DNA molecules at the endogenous FMR1 locus revealed that replication forks stall at CGG repeats in human cells. Furthermore, replication profiles of FXS human embryonic stem cells (hESCs) compared to nonaffected hESCs showed that fork direction through the repeats is altered at the FMR1 locus in FXS hESCs, such that predominantly the CCG strand serves as the lagging-strand template. This is due to the absence of replication initiation that would typically occur upstream of FMR1, suggesting that altered replication origin usage combined with fork stalling promotes repeat instability during early embryonic development.

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The DNA Replication Program Is Altered at the FMR1 Locus in Fragile X Embryonic Stem Cells

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