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Requirement for Atr in phosphorylation of Chk1 and cell cycle regulation in response to DNA replication blocks and UV-damaged DNA in Xenopus egg extracts
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| Zeitschriftentitel: | Genes & Development |
|---|---|
| Personen und Körperschaften: | , , , |
| In: | Genes & Development, 14, 2000, 21, S. 2745-2756 |
| Format: | E-Article |
| Sprache: | Englisch |
| veröffentlicht: |
Cold Spring Harbor Laboratory
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| Schlagwörter: |
| author_facet |
Guo, Zijian Kumagai, Akiko Wang, Sophie X. Dunphy, William G. Guo, Zijian Kumagai, Akiko Wang, Sophie X. Dunphy, William G. |
|---|---|
| author |
Guo, Zijian Kumagai, Akiko Wang, Sophie X. Dunphy, William G. |
| spellingShingle |
Guo, Zijian Kumagai, Akiko Wang, Sophie X. Dunphy, William G. Genes & Development Requirement for Atr in phosphorylation of Chk1 and cell cycle regulation in response to DNA replication blocks and UV-damaged DNA in Xenopus egg extracts Developmental Biology Genetics |
| author_sort |
guo, zijian |
| spelling |
Guo, Zijian Kumagai, Akiko Wang, Sophie X. Dunphy, William G. 0890-9369 1549-5477 Cold Spring Harbor Laboratory Developmental Biology Genetics http://dx.doi.org/10.1101/gad.842500 <jats:p>The checkpoint kinase Xchk1 becomes phosphorylated in<jats:italic>Xenopus</jats:italic> egg extracts in response to DNA replication blocks or UV-damaged DNA. Xchk1 is also required for the cell cycle delay that is induced by unreplicated or UV-damaged DNA. In this report, we have removed the <jats:italic>Xenopus</jats:italic> homolog of ATR (Xatr) from egg extracts by immunodepletion. In Xatr-depleted extracts, the checkpoint-associated phosphorylation of Xchk1 is abolished, and the cell cycle delay induced by replication blocks is strongly compromised. Xatr from egg extracts phosphorylated recombinant Xchk1 in vitro, but not a mutant form of Xchk1 (Xchk1-4AQ) containing nonphosphorylatable residues in its four conserved SQ/TQ motifs. Recombinant human ATR, but not a kinase-inactive mutant, phosphorylated the same sites in Xchk1. Furthermore, the Xchk1-4AQ mutant was found to be defective in mediating a checkpoint response in egg extracts. These findings suggest that Xchk1 is a functionally important target of Xatr during a checkpoint response to unreplicated or UV-damaged DNA.</jats:p> Requirement for Atr in phosphorylation of Chk1 and cell cycle regulation in response to DNA replication blocks and UV-damaged DNA in <i>Xenopus</i> egg extracts Genes & Development |
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Cold Spring Harbor Laboratory, 2000 |
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Cold Spring Harbor Laboratory, 2000 |
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| title |
Requirement for Atr in phosphorylation of Chk1 and cell cycle regulation in response to DNA replication blocks and UV-damaged DNA in Xenopus egg extracts |
| title_unstemmed |
Requirement for Atr in phosphorylation of Chk1 and cell cycle regulation in response to DNA replication blocks and UV-damaged DNA in Xenopus egg extracts |
| title_full |
Requirement for Atr in phosphorylation of Chk1 and cell cycle regulation in response to DNA replication blocks and UV-damaged DNA in Xenopus egg extracts |
| title_fullStr |
Requirement for Atr in phosphorylation of Chk1 and cell cycle regulation in response to DNA replication blocks and UV-damaged DNA in Xenopus egg extracts |
| title_full_unstemmed |
Requirement for Atr in phosphorylation of Chk1 and cell cycle regulation in response to DNA replication blocks and UV-damaged DNA in Xenopus egg extracts |
| title_short |
Requirement for Atr in phosphorylation of Chk1 and cell cycle regulation in response to DNA replication blocks and UV-damaged DNA in Xenopus egg extracts |
| title_sort |
requirement for atr in phosphorylation of chk1 and cell cycle regulation in response to dna replication blocks and uv-damaged dna in <i>xenopus</i> egg extracts |
| topic |
Developmental Biology Genetics |
| url |
http://dx.doi.org/10.1101/gad.842500 |
| publishDate |
2000 |
| physical |
2745-2756 |
| description |
<jats:p>The checkpoint kinase Xchk1 becomes phosphorylated in<jats:italic>Xenopus</jats:italic> egg extracts in response to DNA replication blocks or UV-damaged DNA. Xchk1 is also required for the cell cycle delay that is induced by unreplicated or UV-damaged DNA. In this report, we have removed the <jats:italic>Xenopus</jats:italic> homolog of ATR (Xatr) from egg extracts by immunodepletion. In Xatr-depleted extracts, the checkpoint-associated phosphorylation of Xchk1 is abolished, and the cell cycle delay induced by replication blocks is strongly compromised. Xatr from egg extracts phosphorylated recombinant Xchk1 in vitro, but not a mutant form of Xchk1 (Xchk1-4AQ) containing nonphosphorylatable residues in its four conserved SQ/TQ motifs. Recombinant human ATR, but not a kinase-inactive mutant, phosphorylated the same sites in Xchk1. Furthermore, the Xchk1-4AQ mutant was found to be defective in mediating a checkpoint response in egg extracts. These findings suggest that Xchk1 is a functionally important target of Xatr during a checkpoint response to unreplicated or UV-damaged DNA.</jats:p> |
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| author | Guo, Zijian, Kumagai, Akiko, Wang, Sophie X., Dunphy, William G. |
| author_facet | Guo, Zijian, Kumagai, Akiko, Wang, Sophie X., Dunphy, William G., Guo, Zijian, Kumagai, Akiko, Wang, Sophie X., Dunphy, William G. |
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| description | <jats:p>The checkpoint kinase Xchk1 becomes phosphorylated in<jats:italic>Xenopus</jats:italic> egg extracts in response to DNA replication blocks or UV-damaged DNA. Xchk1 is also required for the cell cycle delay that is induced by unreplicated or UV-damaged DNA. In this report, we have removed the <jats:italic>Xenopus</jats:italic> homolog of ATR (Xatr) from egg extracts by immunodepletion. In Xatr-depleted extracts, the checkpoint-associated phosphorylation of Xchk1 is abolished, and the cell cycle delay induced by replication blocks is strongly compromised. Xatr from egg extracts phosphorylated recombinant Xchk1 in vitro, but not a mutant form of Xchk1 (Xchk1-4AQ) containing nonphosphorylatable residues in its four conserved SQ/TQ motifs. Recombinant human ATR, but not a kinase-inactive mutant, phosphorylated the same sites in Xchk1. Furthermore, the Xchk1-4AQ mutant was found to be defective in mediating a checkpoint response in egg extracts. These findings suggest that Xchk1 is a functionally important target of Xatr during a checkpoint response to unreplicated or UV-damaged DNA.</jats:p> |
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| institution | DE-D275, DE-Bn3, DE-Brt1, DE-D161, DE-Zwi2, DE-Gla1, DE-Zi4, DE-15, DE-Pl11, DE-Rs1, DE-105, DE-14, DE-Ch1, DE-L229 |
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| spelling | Guo, Zijian Kumagai, Akiko Wang, Sophie X. Dunphy, William G. 0890-9369 1549-5477 Cold Spring Harbor Laboratory Developmental Biology Genetics http://dx.doi.org/10.1101/gad.842500 <jats:p>The checkpoint kinase Xchk1 becomes phosphorylated in<jats:italic>Xenopus</jats:italic> egg extracts in response to DNA replication blocks or UV-damaged DNA. Xchk1 is also required for the cell cycle delay that is induced by unreplicated or UV-damaged DNA. In this report, we have removed the <jats:italic>Xenopus</jats:italic> homolog of ATR (Xatr) from egg extracts by immunodepletion. In Xatr-depleted extracts, the checkpoint-associated phosphorylation of Xchk1 is abolished, and the cell cycle delay induced by replication blocks is strongly compromised. Xatr from egg extracts phosphorylated recombinant Xchk1 in vitro, but not a mutant form of Xchk1 (Xchk1-4AQ) containing nonphosphorylatable residues in its four conserved SQ/TQ motifs. Recombinant human ATR, but not a kinase-inactive mutant, phosphorylated the same sites in Xchk1. Furthermore, the Xchk1-4AQ mutant was found to be defective in mediating a checkpoint response in egg extracts. These findings suggest that Xchk1 is a functionally important target of Xatr during a checkpoint response to unreplicated or UV-damaged DNA.</jats:p> Requirement for Atr in phosphorylation of Chk1 and cell cycle regulation in response to DNA replication blocks and UV-damaged DNA in <i>Xenopus</i> egg extracts Genes & Development |
| spellingShingle | Guo, Zijian, Kumagai, Akiko, Wang, Sophie X., Dunphy, William G., Genes & Development, Requirement for Atr in phosphorylation of Chk1 and cell cycle regulation in response to DNA replication blocks and UV-damaged DNA in Xenopus egg extracts, Developmental Biology, Genetics |
| title | Requirement for Atr in phosphorylation of Chk1 and cell cycle regulation in response to DNA replication blocks and UV-damaged DNA in Xenopus egg extracts |
| title_full | Requirement for Atr in phosphorylation of Chk1 and cell cycle regulation in response to DNA replication blocks and UV-damaged DNA in Xenopus egg extracts |
| title_fullStr | Requirement for Atr in phosphorylation of Chk1 and cell cycle regulation in response to DNA replication blocks and UV-damaged DNA in Xenopus egg extracts |
| title_full_unstemmed | Requirement for Atr in phosphorylation of Chk1 and cell cycle regulation in response to DNA replication blocks and UV-damaged DNA in Xenopus egg extracts |
| title_short | Requirement for Atr in phosphorylation of Chk1 and cell cycle regulation in response to DNA replication blocks and UV-damaged DNA in Xenopus egg extracts |
| title_sort | requirement for atr in phosphorylation of chk1 and cell cycle regulation in response to dna replication blocks and uv-damaged dna in <i>xenopus</i> egg extracts |
| title_unstemmed | Requirement for Atr in phosphorylation of Chk1 and cell cycle regulation in response to DNA replication blocks and UV-damaged DNA in Xenopus egg extracts |
| topic | Developmental Biology, Genetics |
| url | http://dx.doi.org/10.1101/gad.842500 |