Damage and replication checkpoint control in fission yeast is ensured by interactions of Crb2, a protein with BRCT motif, with Cut5 and Chk1

Yasushi Saka, F Esashi, T Matsusaka, S Mochida, M Yanagida

Research output: Contribution to journalArticle

219 Citations (Scopus)

Abstract

Fission yeast Cut5/Rad4 plays a unique role in the genome maintenance as it is required for replication, replication checkpoint, and normal UV sensitivity. It is unknown, however, how Cut5 protein is linked to other checkpoint proteins, and what part it plays in replication and UV sensitivity. Here we report that Cut5 interacts with a novel checkpoint protein Crb2 and that this interaction is needed for normal genome maintenance. The carboxyl terminus of Crb2 resembles yeast Rad9 and human 53BP1 and BRCA1. Crb2 is required for checkpoint arrests induced by irradiation and polymerase mutations, but not for those induced by inhibited nucleotide supply. Upon UV damage, Crb2 is transiently modified, probably phosphorylated, with a similar timing of phosphorylation in Chk1 kinase, which is reported to restrain Cdc2 activation. Crb2 modification requires other damage-sensing checkpoint proteins but not Chk1, suggesting that Crb2 acts at the upstream of Chk1. The modified Crb2 exists as a slowly sedimenting form, whereas Crb2 in undamaged cells is in a rapidly sedimenting structure. Cut5 and Crb2 interact with Chk1 in a two-hybrid system. Moreover, moderate overexpression of Chk1 suppresses the phenotypes of cut5 and crb2 mutants. Cut5, Crb2, and Chk1 thus may form a checkpoint sensor-transmitter pathway to arrest the cell cycle.
Original languageEnglish
Pages (from-to)3387-3400
Number of pages14
JournalGenes & Development
Volume11
Issue number24
DOIs
Publication statusPublished - 15 Dec 1997

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Schizosaccharomyces
Proteins
Maintenance
Genome
Cell Cycle Checkpoints
Nucleotides
Yeasts
Phosphorylation
Phenotype
Mutation

Keywords

  • Amino Acid Sequence
  • BRCA1 Protein
  • Binding Sites
  • Cell Cycle Proteins
  • Conserved Sequence
  • DNA Damage
  • DNA Replication
  • DNA-Binding Proteins
  • DNA-Directed DNA Polymerase
  • Fungal Proteins
  • Genes, Fungal
  • Hydroxyurea
  • Molecular Sequence Data
  • Mutation
  • Nuclear Proteins
  • Protein Kinases
  • Repetitive Sequences, Nucleic Acid
  • Schizosaccharomyces
  • Schizosaccharomyces pombe Proteins
  • Sequence Homology, Amino Acid
  • Transglutaminases
  • Ultraviolet Rays
  • UV damage
  • hydroxyurea
  • DNA polymerase
  • phosphorylation
  • two-hybrid screen

Cite this

Damage and replication checkpoint control in fission yeast is ensured by interactions of Crb2, a protein with BRCT motif, with Cut5 and Chk1. / Saka, Yasushi; Esashi, F; Matsusaka, T; Mochida, S; Yanagida, M.

In: Genes & Development, Vol. 11, No. 24, 15.12.1997, p. 3387-3400.

Research output: Contribution to journalArticle

Saka, Yasushi ; Esashi, F ; Matsusaka, T ; Mochida, S ; Yanagida, M. / Damage and replication checkpoint control in fission yeast is ensured by interactions of Crb2, a protein with BRCT motif, with Cut5 and Chk1. In: Genes & Development. 1997 ; Vol. 11, No. 24. pp. 3387-3400.
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abstract = "Fission yeast Cut5/Rad4 plays a unique role in the genome maintenance as it is required for replication, replication checkpoint, and normal UV sensitivity. It is unknown, however, how Cut5 protein is linked to other checkpoint proteins, and what part it plays in replication and UV sensitivity. Here we report that Cut5 interacts with a novel checkpoint protein Crb2 and that this interaction is needed for normal genome maintenance. The carboxyl terminus of Crb2 resembles yeast Rad9 and human 53BP1 and BRCA1. Crb2 is required for checkpoint arrests induced by irradiation and polymerase mutations, but not for those induced by inhibited nucleotide supply. Upon UV damage, Crb2 is transiently modified, probably phosphorylated, with a similar timing of phosphorylation in Chk1 kinase, which is reported to restrain Cdc2 activation. Crb2 modification requires other damage-sensing checkpoint proteins but not Chk1, suggesting that Crb2 acts at the upstream of Chk1. The modified Crb2 exists as a slowly sedimenting form, whereas Crb2 in undamaged cells is in a rapidly sedimenting structure. Cut5 and Crb2 interact with Chk1 in a two-hybrid system. Moreover, moderate overexpression of Chk1 suppresses the phenotypes of cut5 and crb2 mutants. Cut5, Crb2, and Chk1 thus may form a checkpoint sensor-transmitter pathway to arrest the cell cycle.",
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T1 - Damage and replication checkpoint control in fission yeast is ensured by interactions of Crb2, a protein with BRCT motif, with Cut5 and Chk1

AU - Saka, Yasushi

AU - Esashi, F

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AU - Mochida, S

AU - Yanagida, M

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N2 - Fission yeast Cut5/Rad4 plays a unique role in the genome maintenance as it is required for replication, replication checkpoint, and normal UV sensitivity. It is unknown, however, how Cut5 protein is linked to other checkpoint proteins, and what part it plays in replication and UV sensitivity. Here we report that Cut5 interacts with a novel checkpoint protein Crb2 and that this interaction is needed for normal genome maintenance. The carboxyl terminus of Crb2 resembles yeast Rad9 and human 53BP1 and BRCA1. Crb2 is required for checkpoint arrests induced by irradiation and polymerase mutations, but not for those induced by inhibited nucleotide supply. Upon UV damage, Crb2 is transiently modified, probably phosphorylated, with a similar timing of phosphorylation in Chk1 kinase, which is reported to restrain Cdc2 activation. Crb2 modification requires other damage-sensing checkpoint proteins but not Chk1, suggesting that Crb2 acts at the upstream of Chk1. The modified Crb2 exists as a slowly sedimenting form, whereas Crb2 in undamaged cells is in a rapidly sedimenting structure. Cut5 and Crb2 interact with Chk1 in a two-hybrid system. Moreover, moderate overexpression of Chk1 suppresses the phenotypes of cut5 and crb2 mutants. Cut5, Crb2, and Chk1 thus may form a checkpoint sensor-transmitter pathway to arrest the cell cycle.

AB - Fission yeast Cut5/Rad4 plays a unique role in the genome maintenance as it is required for replication, replication checkpoint, and normal UV sensitivity. It is unknown, however, how Cut5 protein is linked to other checkpoint proteins, and what part it plays in replication and UV sensitivity. Here we report that Cut5 interacts with a novel checkpoint protein Crb2 and that this interaction is needed for normal genome maintenance. The carboxyl terminus of Crb2 resembles yeast Rad9 and human 53BP1 and BRCA1. Crb2 is required for checkpoint arrests induced by irradiation and polymerase mutations, but not for those induced by inhibited nucleotide supply. Upon UV damage, Crb2 is transiently modified, probably phosphorylated, with a similar timing of phosphorylation in Chk1 kinase, which is reported to restrain Cdc2 activation. Crb2 modification requires other damage-sensing checkpoint proteins but not Chk1, suggesting that Crb2 acts at the upstream of Chk1. The modified Crb2 exists as a slowly sedimenting form, whereas Crb2 in undamaged cells is in a rapidly sedimenting structure. Cut5 and Crb2 interact with Chk1 in a two-hybrid system. Moreover, moderate overexpression of Chk1 suppresses the phenotypes of cut5 and crb2 mutants. Cut5, Crb2, and Chk1 thus may form a checkpoint sensor-transmitter pathway to arrest the cell cycle.

KW - Amino Acid Sequence

KW - BRCA1 Protein

KW - Binding Sites

KW - Cell Cycle Proteins

KW - Conserved Sequence

KW - DNA Damage

KW - DNA Replication

KW - DNA-Binding Proteins

KW - DNA-Directed DNA Polymerase

KW - Fungal Proteins

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KW - Protein Kinases

KW - Repetitive Sequences, Nucleic Acid

KW - Schizosaccharomyces

KW - Schizosaccharomyces pombe Proteins

KW - Sequence Homology, Amino Acid

KW - Transglutaminases

KW - Ultraviolet Rays

KW - UV damage

KW - hydroxyurea

KW - DNA polymerase

KW - phosphorylation

KW - two-hybrid screen

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JO - Genes & Development

JF - Genes & Development

SN - 0890-9369

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