Fanconi anemia-associated chromosomal radial formation is dependent on POLθ-mediated alternative end joining

Colette B. Rogers; Rachel E. Kram; Kevin Lin; Chad L. Myers; Alexandra Sobeck; Eric A. Hendrickson; Anja Katrin Bielinsky

doi:10.1016/j.celrep.2023.112428

Fanconi anemia-associated chromosomal radial formation is dependent on POLθ-mediated alternative end joining

Colette B. Rogers, Rachel E. Kram, Kevin Lin, Chad L. Myers, Alexandra Sobeck, Eric A. Hendrickson, Anja Katrin Bielinsky

Research output: Contribution to journal › Article › peer-review

1 Scopus citations

Abstract

Activation of the Fanconi anemia (FA) pathway after treatment with mitomycin C (MMC) is essential for preventing chromosome translocations termed “radials.” When replication forks stall at MMC-induced interstrand crosslinks (ICLs), the FA pathway is activated to orchestrate ICL unhooking and repair of the DNA break intermediates. However, in FA-deficient cells, how ICL-associated breaks are resolved in a manner that leads to radials is unclear. Here, we demonstrate that MMC-induced radials are dependent on DNA polymerase theta (POLθ)-mediated alternative end joining (A-EJ). Specifically, we show that radials observed in FANCD2^−/− cells are dependent on POLθ and DNA ligase III and occur independently of classical non-homologous end joining. Furthermore, treatment of FANCD2^−/− cells with POLθ inhibitors abolishes radials and leads to the accumulation of breaks co-localizing with common fragile sites. Uniformly, these observations implicate A-EJ in radial formation and provide mechanistic insights into the treatment of FA pathway-deficient cancers with POLθ inhibitors.

Original language	English (US)
Article number	112428
Journal	Cell reports
Volume	42
Issue number	5
DOIs	https://doi.org/10.1016/j.celrep.2023.112428
State	Published - May 30 2023

Bibliographical note

Funding Information:
We would like to thank the University of Minnesota Cancer Genomics Shared Resource (funded by P30CA077598 CCSG) and the University of Minnesota Imaging Centers for their invaluable assistance. We thank members of the Friday afternoon 3R club for critical discussions and helpful analyses. NIH grants that supported this study are acknowledged as follows: R01GM134681 and R35GM141805 to A.-K.B., R01CA266524 to E.A.H., R01GM132596 to A.S., R01HG005084 to C.L.M., T32CA009138 to C.B.R., and F30CA257227 to K.L.

Publisher Copyright:
© 2023 The Author(s)

Keywords

CP: Molecular biology
FANCD2
Fanconi anemia
POLθ
RAD18
alternative end joining
common fragile sites
double-stranded break repair
radial chromosomes

PubMed: MeSH publication types

Journal Article
Research Support, N.I.H., Extramural

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

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10.1016/j.celrep.2023.112428

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@article{ccabbdb082544e45b056f8c9693b21af,

title = "Fanconi anemia-associated chromosomal radial formation is dependent on POLθ-mediated alternative end joining",

abstract = "Activation of the Fanconi anemia (FA) pathway after treatment with mitomycin C (MMC) is essential for preventing chromosome translocations termed “radials.” When replication forks stall at MMC-induced interstrand crosslinks (ICLs), the FA pathway is activated to orchestrate ICL unhooking and repair of the DNA break intermediates. However, in FA-deficient cells, how ICL-associated breaks are resolved in a manner that leads to radials is unclear. Here, we demonstrate that MMC-induced radials are dependent on DNA polymerase theta (POLθ)-mediated alternative end joining (A-EJ). Specifically, we show that radials observed in FANCD2−/− cells are dependent on POLθ and DNA ligase III and occur independently of classical non-homologous end joining. Furthermore, treatment of FANCD2−/− cells with POLθ inhibitors abolishes radials and leads to the accumulation of breaks co-localizing with common fragile sites. Uniformly, these observations implicate A-EJ in radial formation and provide mechanistic insights into the treatment of FA pathway-deficient cancers with POLθ inhibitors.",

keywords = "CP: Molecular biology, FANCD2, Fanconi anemia, POLθ, RAD18, alternative end joining, common fragile sites, double-stranded break repair, radial chromosomes",

author = "Rogers, {Colette B.} and Kram, {Rachel E.} and Kevin Lin and Myers, {Chad L.} and Alexandra Sobeck and Hendrickson, {Eric A.} and Bielinsky, {Anja Katrin}",

note = "Funding Information: We would like to thank the University of Minnesota Cancer Genomics Shared Resource (funded by P30CA077598 CCSG) and the University of Minnesota Imaging Centers for their invaluable assistance. We thank members of the Friday afternoon 3R club for critical discussions and helpful analyses. NIH grants that supported this study are acknowledged as follows: R01GM134681 and R35GM141805 to A.-K.B., R01CA266524 to E.A.H., R01GM132596 to A.S., R01HG005084 to C.L.M., T32CA009138 to C.B.R., and F30CA257227 to K.L. Publisher Copyright: {\textcopyright} 2023 The Author(s)",

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doi = "10.1016/j.celrep.2023.112428",

language = "English (US)",

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T1 - Fanconi anemia-associated chromosomal radial formation is dependent on POLθ-mediated alternative end joining

AU - Rogers, Colette B.

AU - Kram, Rachel E.

AU - Lin, Kevin

AU - Myers, Chad L.

AU - Sobeck, Alexandra

AU - Hendrickson, Eric A.

AU - Bielinsky, Anja Katrin

N1 - Funding Information: We would like to thank the University of Minnesota Cancer Genomics Shared Resource (funded by P30CA077598 CCSG) and the University of Minnesota Imaging Centers for their invaluable assistance. We thank members of the Friday afternoon 3R club for critical discussions and helpful analyses. NIH grants that supported this study are acknowledged as follows: R01GM134681 and R35GM141805 to A.-K.B., R01CA266524 to E.A.H., R01GM132596 to A.S., R01HG005084 to C.L.M., T32CA009138 to C.B.R., and F30CA257227 to K.L. Publisher Copyright: © 2023 The Author(s)

PY - 2023/5/30

Y1 - 2023/5/30

N2 - Activation of the Fanconi anemia (FA) pathway after treatment with mitomycin C (MMC) is essential for preventing chromosome translocations termed “radials.” When replication forks stall at MMC-induced interstrand crosslinks (ICLs), the FA pathway is activated to orchestrate ICL unhooking and repair of the DNA break intermediates. However, in FA-deficient cells, how ICL-associated breaks are resolved in a manner that leads to radials is unclear. Here, we demonstrate that MMC-induced radials are dependent on DNA polymerase theta (POLθ)-mediated alternative end joining (A-EJ). Specifically, we show that radials observed in FANCD2−/− cells are dependent on POLθ and DNA ligase III and occur independently of classical non-homologous end joining. Furthermore, treatment of FANCD2−/− cells with POLθ inhibitors abolishes radials and leads to the accumulation of breaks co-localizing with common fragile sites. Uniformly, these observations implicate A-EJ in radial formation and provide mechanistic insights into the treatment of FA pathway-deficient cancers with POLθ inhibitors.

AB - Activation of the Fanconi anemia (FA) pathway after treatment with mitomycin C (MMC) is essential for preventing chromosome translocations termed “radials.” When replication forks stall at MMC-induced interstrand crosslinks (ICLs), the FA pathway is activated to orchestrate ICL unhooking and repair of the DNA break intermediates. However, in FA-deficient cells, how ICL-associated breaks are resolved in a manner that leads to radials is unclear. Here, we demonstrate that MMC-induced radials are dependent on DNA polymerase theta (POLθ)-mediated alternative end joining (A-EJ). Specifically, we show that radials observed in FANCD2−/− cells are dependent on POLθ and DNA ligase III and occur independently of classical non-homologous end joining. Furthermore, treatment of FANCD2−/− cells with POLθ inhibitors abolishes radials and leads to the accumulation of breaks co-localizing with common fragile sites. Uniformly, these observations implicate A-EJ in radial formation and provide mechanistic insights into the treatment of FA pathway-deficient cancers with POLθ inhibitors.

KW - CP: Molecular biology

KW - FANCD2

KW - Fanconi anemia

KW - POLθ

KW - RAD18

KW - alternative end joining

KW - common fragile sites

KW - double-stranded break repair

KW - radial chromosomes

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UR - http://www.scopus.com/inward/citedby.url?scp=85152885498&partnerID=8YFLogxK

U2 - 10.1016/j.celrep.2023.112428

DO - 10.1016/j.celrep.2023.112428

M3 - Article

C2 - 37086407

AN - SCOPUS:85152885498

SN - 2211-1247

VL - 42

JO - Cell reports

JF - Cell reports

IS - 5

M1 - 112428

ER -

Fanconi anemia-associated chromosomal radial formation is dependent on POLθ-mediated alternative end joining

Abstract

Bibliographical note

Keywords

PubMed: MeSH publication types

UN SDGs

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Fanconi anemia-associated chromosomal radial formation is dependent on POLθ-mediated alternative end joining

Abstract

Bibliographical note

Keywords

PubMed: MeSH publication types

UN SDGs

Publisher link

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Other files and links

Fingerprint

University Assets

Zeiss Cell Observer SD Spinning Disk Confocal

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