Chk1 phosphorylation at Ser286 and Ser301 occurs with both stalled DNA replication and damage checkpoint stimulation
We previously reported Chk1 to be phosphorylated at Ser286 and Ser301 by cyclin-dependent kinase (Cdk) 1 during mitosis [T. Shiromizu et al., Genes Cells 11 (2006) 477–485]. Here, we demonstrated that Chk1-Ser286 and -Ser301 phosphorylation also occurs in hydroxyurea (HU)-treated or ultraviolet (UV)...
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| creator | Ikegami, Yosuke Goto, Hidemasa Kiyono, Tohru Enomoto, Masato Kasahara, Kousuke Tomono, Yasuko Tozawa, Keiichi Morita, Akimichi Kohri, Kenjiro Inagaki, Masaki |
| description | We previously reported Chk1 to be phosphorylated at Ser286 and Ser301 by cyclin-dependent kinase (Cdk) 1 during mitosis [T. Shiromizu et al., Genes Cells 11 (2006) 477–485]. Here, we demonstrated that Chk1-Ser286 and -Ser301 phosphorylation also occurs in hydroxyurea (HU)-treated or ultraviolet (UV)-irradiated cells. Unlike the mitosis case, however, Chk1 was phosphorylated not only at Ser286 and Ser301 but also at Ser317 and Ser345 in the checkpoint response. Treatment with Cdk inhibitors diminished Chk1 phosphorylation at Ser286 and Ser301 but not at Ser317 and Ser345 with the latter. In vitro analyses revealed Ser286 and Ser301 on Chk1 to serve as two major phosphorylation sites for Cdk2. Immunoprecipitation analyses further demonstrated that Ser286/Ser301 and Ser317/Ser345 phosphorylation occur in the same Chk1 molecule during the checkpoint response. In addition, Ser286/Ser301 phosphorylation by Cdk2 was observed in Chk1 mutated to Ala at Ser317 and Ser345 (S317A/S345A), as well as Ser317/Ser345 phosphorylation by ATR was in S286A/S301A. Therefore, Chk1 phosphorylation in the checkpoint response is regulated not only by ATR but also by Cdk2. |
| doi_str_mv | 10.1016/j.bbrc.2008.10.119 |
| format | Article |
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Shiromizu et al., Genes Cells 11 (2006) 477–485]. Here, we demonstrated that Chk1-Ser286 and -Ser301 phosphorylation also occurs in hydroxyurea (HU)-treated or ultraviolet (UV)-irradiated cells. Unlike the mitosis case, however, Chk1 was phosphorylated not only at Ser286 and Ser301 but also at Ser317 and Ser345 in the checkpoint response. Treatment with Cdk inhibitors diminished Chk1 phosphorylation at Ser286 and Ser301 but not at Ser317 and Ser345 with the latter. In vitro analyses revealed Ser286 and Ser301 on Chk1 to serve as two major phosphorylation sites for Cdk2. Immunoprecipitation analyses further demonstrated that Ser286/Ser301 and Ser317/Ser345 phosphorylation occur in the same Chk1 molecule during the checkpoint response. In addition, Ser286/Ser301 phosphorylation by Cdk2 was observed in Chk1 mutated to Ala at Ser317 and Ser345 (S317A/S345A), as well as Ser317/Ser345 phosphorylation by ATR was in S286A/S301A. Therefore, Chk1 phosphorylation in the checkpoint response is regulated not only by ATR but also by Cdk2.</description><identifier>ISSN: 0006-291X</identifier><identifier>EISSN: 1090-2104</identifier><identifier>DOI: 10.1016/j.bbrc.2008.10.119</identifier><identifier>PMID: 18983824</identifier><language>eng</language><publisher>United States: Elsevier Inc</publisher><subject>60 APPLIED LIFE SCIENCES ; ATR ; Checkpoint ; Checkpoint Kinase 1 ; Chk1 ; Cyclin-dependent kinase 2 (Cdk2) ; DAMAGE ; DNA - drug effects ; DNA - radiation effects ; DNA Damage ; DNA REPLICATION ; DNA Replication - drug effects ; DNA Replication - radiation effects ; ELECTROPHORESIS ; GELS ; GENES ; GLUTATHIONE ; HeLa Cells ; Humans ; HYDROXYUREA ; Hydroxyurea - pharmacology ; Immunoprecipitation ; IN VITRO ; IRRADIATION ; MITOSIS ; PHOSPHORYLATION ; Protein Kinases - genetics ; Protein Kinases - metabolism ; Serine - genetics ; Serine - metabolism ; STIMULATION ; ULTRAVIOLET RADIATION ; Ultraviolet Rays</subject><ispartof>Biochemical and biophysical research communications, 2008-12, Vol.377 (4), p.1227-1231</ispartof><rights>2008 Elsevier Inc.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c413t-37918696de11b1ee9350b4066ab145823375f242115f2988347f34fb90d14ed53</citedby><cites>FETCH-LOGICAL-c413t-37918696de11b1ee9350b4066ab145823375f242115f2988347f34fb90d14ed53</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://dx.doi.org/10.1016/j.bbrc.2008.10.119$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>230,314,780,784,885,3550,27924,27925,45995</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/18983824$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink><backlink>$$Uhttps://www.osti.gov/biblio/21255809$$D View this record in Osti.gov$$Hfree_for_read</backlink></links><search><creatorcontrib>Ikegami, Yosuke</creatorcontrib><creatorcontrib>Goto, Hidemasa</creatorcontrib><creatorcontrib>Kiyono, Tohru</creatorcontrib><creatorcontrib>Enomoto, Masato</creatorcontrib><creatorcontrib>Kasahara, Kousuke</creatorcontrib><creatorcontrib>Tomono, Yasuko</creatorcontrib><creatorcontrib>Tozawa, Keiichi</creatorcontrib><creatorcontrib>Morita, Akimichi</creatorcontrib><creatorcontrib>Kohri, Kenjiro</creatorcontrib><creatorcontrib>Inagaki, Masaki</creatorcontrib><title>Chk1 phosphorylation at Ser286 and Ser301 occurs with both stalled DNA replication and damage checkpoint stimulation</title><title>Biochemical and biophysical research communications</title><addtitle>Biochem Biophys Res Commun</addtitle><description>We previously reported Chk1 to be phosphorylated at Ser286 and Ser301 by cyclin-dependent kinase (Cdk) 1 during mitosis [T. Shiromizu et al., Genes Cells 11 (2006) 477–485]. Here, we demonstrated that Chk1-Ser286 and -Ser301 phosphorylation also occurs in hydroxyurea (HU)-treated or ultraviolet (UV)-irradiated cells. Unlike the mitosis case, however, Chk1 was phosphorylated not only at Ser286 and Ser301 but also at Ser317 and Ser345 in the checkpoint response. Treatment with Cdk inhibitors diminished Chk1 phosphorylation at Ser286 and Ser301 but not at Ser317 and Ser345 with the latter. In vitro analyses revealed Ser286 and Ser301 on Chk1 to serve as two major phosphorylation sites for Cdk2. Immunoprecipitation analyses further demonstrated that Ser286/Ser301 and Ser317/Ser345 phosphorylation occur in the same Chk1 molecule during the checkpoint response. In addition, Ser286/Ser301 phosphorylation by Cdk2 was observed in Chk1 mutated to Ala at Ser317 and Ser345 (S317A/S345A), as well as Ser317/Ser345 phosphorylation by ATR was in S286A/S301A. Therefore, Chk1 phosphorylation in the checkpoint response is regulated not only by ATR but also by Cdk2.</description><subject>60 APPLIED LIFE SCIENCES</subject><subject>ATR</subject><subject>Checkpoint</subject><subject>Checkpoint Kinase 1</subject><subject>Chk1</subject><subject>Cyclin-dependent kinase 2 (Cdk2)</subject><subject>DAMAGE</subject><subject>DNA - drug effects</subject><subject>DNA - radiation effects</subject><subject>DNA Damage</subject><subject>DNA REPLICATION</subject><subject>DNA Replication - drug effects</subject><subject>DNA Replication - radiation effects</subject><subject>ELECTROPHORESIS</subject><subject>GELS</subject><subject>GENES</subject><subject>GLUTATHIONE</subject><subject>HeLa Cells</subject><subject>Humans</subject><subject>HYDROXYUREA</subject><subject>Hydroxyurea - pharmacology</subject><subject>Immunoprecipitation</subject><subject>IN VITRO</subject><subject>IRRADIATION</subject><subject>MITOSIS</subject><subject>PHOSPHORYLATION</subject><subject>Protein Kinases - genetics</subject><subject>Protein Kinases - metabolism</subject><subject>Serine - genetics</subject><subject>Serine - metabolism</subject><subject>STIMULATION</subject><subject>ULTRAVIOLET RADIATION</subject><subject>Ultraviolet Rays</subject><issn>0006-291X</issn><issn>1090-2104</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2008</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkc-r1DAQx4MovnX1H_AgAcFb10ySdhPw8lh_wkMPKngLaTq12dc2NWmV99-b2gVvekgmDJ_5Tma-hDwFdgAG1cvzoa6jO3DG1GHNgb5HdsA0KzgweZ_sGGNVwTV8uyKPUjozBiAr_ZBcgdJKKC53ZD51t0CnLqR84l1vZx9Gamf6GSNXFbVjsz4FAxqcW2Kiv_zc0TrkK82277Ghrz9e04hT792lOtc0drDfkboO3e0U_Dhn2g_Lpv-YPGhtn_DJJe7J17dvvpzeFzef3n04Xd8UToKYC3HUoCpdNQhQA6IWJaslqypbgywVF-JYtlxygBy0UkIeWyHbWrMGJDal2JPnm27IzU1yfkbXuTCO6GbDgZelYjpTLzZqiuHHgmk2g08O-96OGJZkKq0kE0z-FwR9rEqRN7snfANdDClFbM0U_WDjnQFmVuvM2azWmdW6PzlYv_Hsor7UAzZ_Sy5eZeDVBmBe2U-PcZ0IR4eNj-tATfD_0v8NK76oFA</recordid><startdate>20081226</startdate><enddate>20081226</enddate><creator>Ikegami, Yosuke</creator><creator>Goto, Hidemasa</creator><creator>Kiyono, Tohru</creator><creator>Enomoto, Masato</creator><creator>Kasahara, Kousuke</creator><creator>Tomono, Yasuko</creator><creator>Tozawa, Keiichi</creator><creator>Morita, Akimichi</creator><creator>Kohri, Kenjiro</creator><creator>Inagaki, Masaki</creator><general>Elsevier Inc</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7TM</scope><scope>8FD</scope><scope>FR3</scope><scope>P64</scope><scope>RC3</scope><scope>7X8</scope><scope>OTOTI</scope></search><sort><creationdate>20081226</creationdate><title>Chk1 phosphorylation at Ser286 and Ser301 occurs with both stalled DNA replication and damage checkpoint stimulation</title><author>Ikegami, Yosuke ; Goto, Hidemasa ; Kiyono, Tohru ; Enomoto, Masato ; Kasahara, Kousuke ; Tomono, Yasuko ; Tozawa, Keiichi ; Morita, Akimichi ; Kohri, Kenjiro ; Inagaki, Masaki</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c413t-37918696de11b1ee9350b4066ab145823375f242115f2988347f34fb90d14ed53</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2008</creationdate><topic>60 APPLIED LIFE SCIENCES</topic><topic>ATR</topic><topic>Checkpoint</topic><topic>Checkpoint Kinase 1</topic><topic>Chk1</topic><topic>Cyclin-dependent kinase 2 (Cdk2)</topic><topic>DAMAGE</topic><topic>DNA - drug effects</topic><topic>DNA - radiation effects</topic><topic>DNA Damage</topic><topic>DNA REPLICATION</topic><topic>DNA Replication - drug effects</topic><topic>DNA Replication - radiation effects</topic><topic>ELECTROPHORESIS</topic><topic>GELS</topic><topic>GENES</topic><topic>GLUTATHIONE</topic><topic>HeLa Cells</topic><topic>Humans</topic><topic>HYDROXYUREA</topic><topic>Hydroxyurea - pharmacology</topic><topic>Immunoprecipitation</topic><topic>IN VITRO</topic><topic>IRRADIATION</topic><topic>MITOSIS</topic><topic>PHOSPHORYLATION</topic><topic>Protein Kinases - genetics</topic><topic>Protein Kinases - metabolism</topic><topic>Serine - genetics</topic><topic>Serine - metabolism</topic><topic>STIMULATION</topic><topic>ULTRAVIOLET RADIATION</topic><topic>Ultraviolet Rays</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Ikegami, Yosuke</creatorcontrib><creatorcontrib>Goto, Hidemasa</creatorcontrib><creatorcontrib>Kiyono, Tohru</creatorcontrib><creatorcontrib>Enomoto, Masato</creatorcontrib><creatorcontrib>Kasahara, Kousuke</creatorcontrib><creatorcontrib>Tomono, Yasuko</creatorcontrib><creatorcontrib>Tozawa, Keiichi</creatorcontrib><creatorcontrib>Morita, Akimichi</creatorcontrib><creatorcontrib>Kohri, Kenjiro</creatorcontrib><creatorcontrib>Inagaki, Masaki</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Nucleic Acids Abstracts</collection><collection>Technology Research Database</collection><collection>Engineering Research Database</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>Genetics Abstracts</collection><collection>MEDLINE - Academic</collection><collection>OSTI.GOV</collection><jtitle>Biochemical and biophysical research communications</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Ikegami, Yosuke</au><au>Goto, Hidemasa</au><au>Kiyono, Tohru</au><au>Enomoto, Masato</au><au>Kasahara, Kousuke</au><au>Tomono, Yasuko</au><au>Tozawa, Keiichi</au><au>Morita, Akimichi</au><au>Kohri, Kenjiro</au><au>Inagaki, Masaki</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Chk1 phosphorylation at Ser286 and Ser301 occurs with both stalled DNA replication and damage checkpoint stimulation</atitle><jtitle>Biochemical and biophysical research communications</jtitle><addtitle>Biochem Biophys Res Commun</addtitle><date>2008-12-26</date><risdate>2008</risdate><volume>377</volume><issue>4</issue><spage>1227</spage><epage>1231</epage><pages>1227-1231</pages><issn>0006-291X</issn><eissn>1090-2104</eissn><abstract>We previously reported Chk1 to be phosphorylated at Ser286 and Ser301 by cyclin-dependent kinase (Cdk) 1 during mitosis [T. Shiromizu et al., Genes Cells 11 (2006) 477–485]. Here, we demonstrated that Chk1-Ser286 and -Ser301 phosphorylation also occurs in hydroxyurea (HU)-treated or ultraviolet (UV)-irradiated cells. Unlike the mitosis case, however, Chk1 was phosphorylated not only at Ser286 and Ser301 but also at Ser317 and Ser345 in the checkpoint response. Treatment with Cdk inhibitors diminished Chk1 phosphorylation at Ser286 and Ser301 but not at Ser317 and Ser345 with the latter. In vitro analyses revealed Ser286 and Ser301 on Chk1 to serve as two major phosphorylation sites for Cdk2. Immunoprecipitation analyses further demonstrated that Ser286/Ser301 and Ser317/Ser345 phosphorylation occur in the same Chk1 molecule during the checkpoint response. In addition, Ser286/Ser301 phosphorylation by Cdk2 was observed in Chk1 mutated to Ala at Ser317 and Ser345 (S317A/S345A), as well as Ser317/Ser345 phosphorylation by ATR was in S286A/S301A. Therefore, Chk1 phosphorylation in the checkpoint response is regulated not only by ATR but also by Cdk2.</abstract><cop>United States</cop><pub>Elsevier Inc</pub><pmid>18983824</pmid><doi>10.1016/j.bbrc.2008.10.119</doi><tpages>5</tpages></addata></record> |
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| subjects | 60 APPLIED LIFE SCIENCES ATR Checkpoint Checkpoint Kinase 1 Chk1 Cyclin-dependent kinase 2 (Cdk2) DAMAGE DNA - drug effects DNA - radiation effects DNA Damage DNA REPLICATION DNA Replication - drug effects DNA Replication - radiation effects ELECTROPHORESIS GELS GENES GLUTATHIONE HeLa Cells Humans HYDROXYUREA Hydroxyurea - pharmacology Immunoprecipitation IN VITRO IRRADIATION MITOSIS PHOSPHORYLATION Protein Kinases - genetics Protein Kinases - metabolism Serine - genetics Serine - metabolism STIMULATION ULTRAVIOLET RADIATION Ultraviolet Rays |
| title | Chk1 phosphorylation at Ser286 and Ser301 occurs with both stalled DNA replication and damage checkpoint stimulation |
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