Caenorhabditis elegans BUB-3 and SAN-1/MAD3 Spindle Assembly Checkpoint Components Are Required for Genome Stability in Response to Treatment with Ionizing Radiation

Simone Bertolini, Bin Wang, Bettina Meier, Ye Hong, Anton Gartner (Lead / Corresponding author)

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Abstract

Relatively little is known about the crosstalk between the spindle assembly checkpoint and the DNA damage response, especially in multicellular organisms. We performed a Caenorhabditis elegans forward genetic screen to uncover new genes involved in the repair of DNA damage induced by ionizing radiation. We isolated a mutation, gt2000 which confers hypersensitivity to ionizing radiation and showed that gt2000 introduces a premature stop in bub-3 BUB-3 is a key component of the spindle assembly checkpoint. We provide evidence that BUB-3 acts during development and in the germline; irradiated bub-3(gt2000) larvae are developmentally retarded and form abnormal vulvae. Moreover, bub-3(gt2000) embryos sired from irradiated worms show increased levels of lethality. Both bub-3 and san-1 (the Caenorhabditis elegans homologue of MAD3) deletion alleles confer hypersensitivity to ionizing radiation, consistent with the notion that the spindle assembly checkpoint pathway is required for DNA damage response. bub-3(gt2000) is moderately sensitive to the crosslinking drug cisplatin but not to UV light or methyl methanesulfonate. This is consistent with role in dealing with DNA double-strand breaks and not with base damage. Double mutant analysis revealed that bub-3 does not act within any of the three major pathways involved in the repair of double-strand breaks. Finally, the cdc-20 gain-of-function mutant cdc-20/fzy-1(av15), which is refractory to the cell cycle delay conferred by the spindle checkpoint showed phenotypes similar to bub-3 and san-1 mutants. We speculate that BUB-3 is involved in DNA damage response through regulation of cell cycle timing.

Original languageEnglish
Pages (from-to)3875-3885
Number of pages11
JournalG3 : Genes, Genomes, Genetics
Volume7
Issue number12
Early online date18 Oct 2017
DOIs
Publication statusPublished - 1 Dec 2017

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M Phase Cell Cycle Checkpoints
Genomic Instability
Caenorhabditis elegans
Ionizing Radiation
DNA Damage
Cell Cycle
Hypersensitivity
Methyl Methanesulfonate
Vulva
Double-Stranded DNA Breaks
Ultraviolet Rays
Cisplatin
Larva
Embryonic Structures
Alleles
Phenotype
Mutation
Pharmaceutical Preparations
Genes

Keywords

  • Ionizing radiation
  • Spindle Assembly Checkpoint
  • BUB-3, SAN-1/MAD-3
  • DNA damage response

Cite this

@article{00de51c56a414ffaa8e3fe80b44cd75d,
title = "Caenorhabditis elegans BUB-3 and SAN-1/MAD3 Spindle Assembly Checkpoint Components Are Required for Genome Stability in Response to Treatment with Ionizing Radiation",
abstract = "Relatively little is known about the crosstalk between the spindle assembly checkpoint and the DNA damage response, especially in multicellular organisms. We performed a Caenorhabditis elegans forward genetic screen to uncover new genes involved in the repair of DNA damage induced by ionizing radiation. We isolated a mutation, gt2000 which confers hypersensitivity to ionizing radiation and showed that gt2000 introduces a premature stop in bub-3 BUB-3 is a key component of the spindle assembly checkpoint. We provide evidence that BUB-3 acts during development and in the germline; irradiated bub-3(gt2000) larvae are developmentally retarded and form abnormal vulvae. Moreover, bub-3(gt2000) embryos sired from irradiated worms show increased levels of lethality. Both bub-3 and san-1 (the Caenorhabditis elegans homologue of MAD3) deletion alleles confer hypersensitivity to ionizing radiation, consistent with the notion that the spindle assembly checkpoint pathway is required for DNA damage response. bub-3(gt2000) is moderately sensitive to the crosslinking drug cisplatin but not to UV light or methyl methanesulfonate. This is consistent with role in dealing with DNA double-strand breaks and not with base damage. Double mutant analysis revealed that bub-3 does not act within any of the three major pathways involved in the repair of double-strand breaks. Finally, the cdc-20 gain-of-function mutant cdc-20/fzy-1(av15), which is refractory to the cell cycle delay conferred by the spindle checkpoint showed phenotypes similar to bub-3 and san-1 mutants. We speculate that BUB-3 is involved in DNA damage response through regulation of cell cycle timing.",
keywords = "Ionizing radiation, Spindle Assembly Checkpoint, BUB-3, SAN-1/MAD-3 , DNA damage response",
author = "Simone Bertolini and Bin Wang and Bettina Meier and Ye Hong and Anton Gartner",
note = "This work was funded by a Wellcome Trust Programme grant to AG (0909444/Z/09/Z), together with infrastructure funding from a Wellcome Trust Strategic award (097045/B/11/Z). We acknowledge the Dundee Imaging Facility, which is supported by the Wellcome Trust Technology Platform award (097945/B/11/Z) and the MRC Next Generation Optical Microscopy award (MR/K015869/1). BW and YH were also supported by ISSF funding from the Wellcome Trust. SB was supported by a BBSRC EASTBIO PhD studentship.",
year = "2017",
month = "12",
day = "1",
doi = "10.1534/g3.117.1122",
language = "English",
volume = "7",
pages = "3875--3885",
journal = "G3 : Genes, Genomes, Genetics",
issn = "2160-1836",
publisher = "Genetics Society of America",
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TY - JOUR

T1 - Caenorhabditis elegans BUB-3 and SAN-1/MAD3 Spindle Assembly Checkpoint Components Are Required for Genome Stability in Response to Treatment with Ionizing Radiation

AU - Bertolini, Simone

AU - Wang, Bin

AU - Meier, Bettina

AU - Hong, Ye

AU - Gartner, Anton

N1 - This work was funded by a Wellcome Trust Programme grant to AG (0909444/Z/09/Z), together with infrastructure funding from a Wellcome Trust Strategic award (097045/B/11/Z). We acknowledge the Dundee Imaging Facility, which is supported by the Wellcome Trust Technology Platform award (097945/B/11/Z) and the MRC Next Generation Optical Microscopy award (MR/K015869/1). BW and YH were also supported by ISSF funding from the Wellcome Trust. SB was supported by a BBSRC EASTBIO PhD studentship.

PY - 2017/12/1

Y1 - 2017/12/1

N2 - Relatively little is known about the crosstalk between the spindle assembly checkpoint and the DNA damage response, especially in multicellular organisms. We performed a Caenorhabditis elegans forward genetic screen to uncover new genes involved in the repair of DNA damage induced by ionizing radiation. We isolated a mutation, gt2000 which confers hypersensitivity to ionizing radiation and showed that gt2000 introduces a premature stop in bub-3 BUB-3 is a key component of the spindle assembly checkpoint. We provide evidence that BUB-3 acts during development and in the germline; irradiated bub-3(gt2000) larvae are developmentally retarded and form abnormal vulvae. Moreover, bub-3(gt2000) embryos sired from irradiated worms show increased levels of lethality. Both bub-3 and san-1 (the Caenorhabditis elegans homologue of MAD3) deletion alleles confer hypersensitivity to ionizing radiation, consistent with the notion that the spindle assembly checkpoint pathway is required for DNA damage response. bub-3(gt2000) is moderately sensitive to the crosslinking drug cisplatin but not to UV light or methyl methanesulfonate. This is consistent with role in dealing with DNA double-strand breaks and not with base damage. Double mutant analysis revealed that bub-3 does not act within any of the three major pathways involved in the repair of double-strand breaks. Finally, the cdc-20 gain-of-function mutant cdc-20/fzy-1(av15), which is refractory to the cell cycle delay conferred by the spindle checkpoint showed phenotypes similar to bub-3 and san-1 mutants. We speculate that BUB-3 is involved in DNA damage response through regulation of cell cycle timing.

AB - Relatively little is known about the crosstalk between the spindle assembly checkpoint and the DNA damage response, especially in multicellular organisms. We performed a Caenorhabditis elegans forward genetic screen to uncover new genes involved in the repair of DNA damage induced by ionizing radiation. We isolated a mutation, gt2000 which confers hypersensitivity to ionizing radiation and showed that gt2000 introduces a premature stop in bub-3 BUB-3 is a key component of the spindle assembly checkpoint. We provide evidence that BUB-3 acts during development and in the germline; irradiated bub-3(gt2000) larvae are developmentally retarded and form abnormal vulvae. Moreover, bub-3(gt2000) embryos sired from irradiated worms show increased levels of lethality. Both bub-3 and san-1 (the Caenorhabditis elegans homologue of MAD3) deletion alleles confer hypersensitivity to ionizing radiation, consistent with the notion that the spindle assembly checkpoint pathway is required for DNA damage response. bub-3(gt2000) is moderately sensitive to the crosslinking drug cisplatin but not to UV light or methyl methanesulfonate. This is consistent with role in dealing with DNA double-strand breaks and not with base damage. Double mutant analysis revealed that bub-3 does not act within any of the three major pathways involved in the repair of double-strand breaks. Finally, the cdc-20 gain-of-function mutant cdc-20/fzy-1(av15), which is refractory to the cell cycle delay conferred by the spindle checkpoint showed phenotypes similar to bub-3 and san-1 mutants. We speculate that BUB-3 is involved in DNA damage response through regulation of cell cycle timing.

KW - Ionizing radiation

KW - Spindle Assembly Checkpoint

KW - BUB-3, SAN-1/MAD-3

KW - DNA damage response

U2 - 10.1534/g3.117.1122

DO - 10.1534/g3.117.1122

M3 - Article

VL - 7

SP - 3875

EP - 3885

JO - G3 : Genes, Genomes, Genetics

JF - G3 : Genes, Genomes, Genetics

SN - 2160-1836

IS - 12

ER -