@article{959c6bfb20df43ccb27ffa6f81ade3b0,
title = "Sequence and Nuclease Requirements for Breakage and Healing of a Structure-Forming (AT)n Sequence within Fragile Site FRA16D",
abstract = "Common fragile sites (CFSs) are genomic regions that display gaps and breaks in human metaphase chromosomes under replication stress and are often deleted in cancer cells. We studied an ∼300-bp subregion (Flex1) of human CFS FRA16D in yeast and found that it recapitulates characteristics of CFS fragility in human cells. Flex1 fragility is dependent on the ability of a variable-length AT repeat to form a cruciform structure that stalls replication. Fragility at Flex1 is initiated by structure-specific endonuclease Mus81-Mms4 acting together with the Slx1-4/Rad1-10 complex, whereas Yen1 protects Flex1 against breakage. Sae2 is required for healing of Flex1 after breakage. Our study shows that breakage within a CFS can be initiated by nuclease cleavage at forks stalled at DNA structures. Furthermore, our results suggest that CFSs are not just prone to breakage but also are impaired in their ability to heal, and this deleterious combination accounts for their fragility.",
author = "Simran Kaushal and Wollmuth, {Charles E.} and Kohal Das and Hile, {Suzanne E.} and Regan, {Samantha B.} and Barnes, {Ryan P.} and Alice Haouzi and Lee, {Soo Mi} and House, {Nealia C.M.} and Michael Guyumdzhyan and Eckert, {Kristin A.} and Freudenreich, {Catherine H.}",
note = "Funding Information: We thank Brian Lenzmeier for sharing reagents to construct the ADE2 version of the DDRA assay; Francesca Storici for the inducible I-SceI system; Ryan McGinty and Michael Sigouros for help with yeast strain construction, verification, and some assays; and Allen Su, Keerthana Gnanapradeepan, and Adam Snider for help with cloning. Funding was provided by NSF grants MCB1330743 and MCB1817499, NIH grants GM105473 and GM122880, and a Tufts Dean's Fund award (to C.H.F.), an Arnold and Mabel Beckman Foundation award (to C.H.F. and C.E.W.), and a Tufts Graduate Student Research Award (to S.K.). Conceptualization, S.K. and C.H.F.; Methodology, S.K. K.A.E. and C.H.F.; Validation, S.K. K.A.E. and C.H.F.; Formal Analysis, S.K.; Investigation, S.K. C.E.W. K.D. S.B.R. A.H. S.E.H. R.P.B. S.M.L. N.C.M.H. and M.G.; Resources, C.H.F. and K.A.E.; Writing – Original Draft, S.K. and C.H.F.; Writing – Review & Editing, S.K. C.H.F. C.E.W. K.A.E. S.M.L. N.C.M.H. S.B.R. R.P.B. and S.E.H.; Visualization, S.K.; Supervision, S.K. K.A.E. and C.H.F.; Project Administration, C.H.F. and K.A.E.; Funding Acquisition, S.K. C.E.W. K.A.E. and C.H.F. The authors declare no competing interests. Funding Information: We thank Brian Lenzmeier for sharing reagents to construct the ADE2 version of the DDRA assay; Francesca Storici for the inducible I-SceI system; Ryan McGinty and Michael Sigouros for help with yeast strain construction, verification, and some assays; and Allen Su, Keerthana Gnanapradeepan, and Adam Snider for help with cloning. Funding was provided by NSF grants MCB1330743 and MCB1817499 , NIH grants GM105473 and GM122880 , and a Tufts Dean{\textquoteright}s Fund award (to C.H.F.), an Arnold and Mabel Beckman Foundation award (to C.H.F. and C.E.W.), and a Tufts Graduate Student Research Award (to S.K.). Publisher Copyright: {\textcopyright} 2019 The Author(s)",
year = "2019",
month = apr,
day = "23",
doi = "10.1016/j.celrep.2019.03.103",
language = "English (US)",
volume = "27",
pages = "1151--1164.e5",
journal = "Cell Reports",
issn = "2211-1247",
publisher = "Cell Press",
number = "4",
}
