Binding of 14-3-3β but not 14-3-3σ controls the cytoplasmic localization of CDC25B

Binding site preferences of 14-3-3 subtypes and the subcellular localization of CDC25B

Sanae Uchida, Akiko Kuma, Motoaki Ohtsubo, Mari Shimura, Masato Hirata, Hitoshi Nakagama, Tsukasa Matsunaga, Yukihito Ishizaka, Katsumi Yamashita

Research output: Contribution to journalArticle

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Abstract

The dual specificity phosphatase CDC25B positively controls the G2-M transition by activating CDK1/cyclin B. The binding of 14-3-3 to CDC25B has been shown to regulate the subcellular redistribution of CDC25B from the nucleus to the cytoplasm and may be correlated with the G2 checkpoint. We used a FLAG-tagged version of CDC25B to study the differences among the binding sites for the 14-3-3 subtypes, 14-3-3β, 14-3-3ε and 14-3-3σ, and the relationship between subtype binding and the subcellular localization of CDC25B. All three subtypes were found to bind to CDC25B. Site-directed mutagenesis studies revealed that 14-3-3β bound exclusively near serine-309 of CDC25B1, which is within a potential consensus motif for 14-3-3 binding. By contrast, 14-3-3σ bound preferentially to a site around serine-216, and the presence of serine-137 and -309 enhanced the binding. In addition to these binding-site differences, we found that the binding of 14-3-3β drove CDC25B to the cytoplasm and that mutation of serine-309 to alanine completely abolished the cytoplasmic localization of CDC25B. However, co-expression of 14-3-3σ and CDC25B did not affect the subeellular localization of CDC25B. Furthermore, serine-309 of CDC25B was sufficient to produce its cytoplasmic distribution with co-expression of 14-3-3β, even when other putative 14-3-3 binding sites were mutated. 14-3-3ε resembled 14-3-3β with regard to its binding to CDC25B and the control of CDC25B subcellujar localization. The results of the present study indicite that two 14-3-3 subtypes can control the subcellular localization of CDC25B by binding to a specific site and that 14-3-3σ has effects on CDC25B other than the control of its subcellular localization.

Original languageEnglish
Pages (from-to)3011-3020
Number of pages10
JournalJournal of Cell Science
Volume117
Issue number14
DOIs
Publication statusPublished - Jun 15 2004

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Serine
Binding Sites
Cytoplasm
Dual-Specificity Phosphatases
Cyclin B
Site-Directed Mutagenesis
Alanine
Mutation

All Science Journal Classification (ASJC) codes

  • Cell Biology

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Binding of 14-3-3β but not 14-3-3σ controls the cytoplasmic localization of CDC25B : Binding site preferences of 14-3-3 subtypes and the subcellular localization of CDC25B. / Uchida, Sanae; Kuma, Akiko; Ohtsubo, Motoaki; Shimura, Mari; Hirata, Masato; Nakagama, Hitoshi; Matsunaga, Tsukasa; Ishizaka, Yukihito; Yamashita, Katsumi.

In: Journal of Cell Science, Vol. 117, No. 14, 15.06.2004, p. 3011-3020.

Research output: Contribution to journalArticle

Uchida, S, Kuma, A, Ohtsubo, M, Shimura, M, Hirata, M, Nakagama, H, Matsunaga, T, Ishizaka, Y & Yamashita, K 2004, 'Binding of 14-3-3β but not 14-3-3σ controls the cytoplasmic localization of CDC25B: Binding site preferences of 14-3-3 subtypes and the subcellular localization of CDC25B', Journal of Cell Science, vol. 117, no. 14, pp. 3011-3020. https://doi.org/10.1242/jcs.01086
Uchida, Sanae ; Kuma, Akiko ; Ohtsubo, Motoaki ; Shimura, Mari ; Hirata, Masato ; Nakagama, Hitoshi ; Matsunaga, Tsukasa ; Ishizaka, Yukihito ; Yamashita, Katsumi. / Binding of 14-3-3β but not 14-3-3σ controls the cytoplasmic localization of CDC25B : Binding site preferences of 14-3-3 subtypes and the subcellular localization of CDC25B. In: Journal of Cell Science. 2004 ; Vol. 117, No. 14. pp. 3011-3020.
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abstract = "The dual specificity phosphatase CDC25B positively controls the G2-M transition by activating CDK1/cyclin B. The binding of 14-3-3 to CDC25B has been shown to regulate the subcellular redistribution of CDC25B from the nucleus to the cytoplasm and may be correlated with the G2 checkpoint. We used a FLAG-tagged version of CDC25B to study the differences among the binding sites for the 14-3-3 subtypes, 14-3-3β, 14-3-3ε and 14-3-3σ, and the relationship between subtype binding and the subcellular localization of CDC25B. All three subtypes were found to bind to CDC25B. Site-directed mutagenesis studies revealed that 14-3-3β bound exclusively near serine-309 of CDC25B1, which is within a potential consensus motif for 14-3-3 binding. By contrast, 14-3-3σ bound preferentially to a site around serine-216, and the presence of serine-137 and -309 enhanced the binding. In addition to these binding-site differences, we found that the binding of 14-3-3β drove CDC25B to the cytoplasm and that mutation of serine-309 to alanine completely abolished the cytoplasmic localization of CDC25B. However, co-expression of 14-3-3σ and CDC25B did not affect the subeellular localization of CDC25B. Furthermore, serine-309 of CDC25B was sufficient to produce its cytoplasmic distribution with co-expression of 14-3-3β, even when other putative 14-3-3 binding sites were mutated. 14-3-3ε resembled 14-3-3β with regard to its binding to CDC25B and the control of CDC25B subcellujar localization. The results of the present study indicite that two 14-3-3 subtypes can control the subcellular localization of CDC25B by binding to a specific site and that 14-3-3σ has effects on CDC25B other than the control of its subcellular localization.",
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AU - Kuma, Akiko

AU - Ohtsubo, Motoaki

AU - Shimura, Mari

AU - Hirata, Masato

AU - Nakagama, Hitoshi

AU - Matsunaga, Tsukasa

AU - Ishizaka, Yukihito

AU - Yamashita, Katsumi

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AB - The dual specificity phosphatase CDC25B positively controls the G2-M transition by activating CDK1/cyclin B. The binding of 14-3-3 to CDC25B has been shown to regulate the subcellular redistribution of CDC25B from the nucleus to the cytoplasm and may be correlated with the G2 checkpoint. We used a FLAG-tagged version of CDC25B to study the differences among the binding sites for the 14-3-3 subtypes, 14-3-3β, 14-3-3ε and 14-3-3σ, and the relationship between subtype binding and the subcellular localization of CDC25B. All three subtypes were found to bind to CDC25B. Site-directed mutagenesis studies revealed that 14-3-3β bound exclusively near serine-309 of CDC25B1, which is within a potential consensus motif for 14-3-3 binding. By contrast, 14-3-3σ bound preferentially to a site around serine-216, and the presence of serine-137 and -309 enhanced the binding. In addition to these binding-site differences, we found that the binding of 14-3-3β drove CDC25B to the cytoplasm and that mutation of serine-309 to alanine completely abolished the cytoplasmic localization of CDC25B. However, co-expression of 14-3-3σ and CDC25B did not affect the subeellular localization of CDC25B. Furthermore, serine-309 of CDC25B was sufficient to produce its cytoplasmic distribution with co-expression of 14-3-3β, even when other putative 14-3-3 binding sites were mutated. 14-3-3ε resembled 14-3-3β with regard to its binding to CDC25B and the control of CDC25B subcellujar localization. The results of the present study indicite that two 14-3-3 subtypes can control the subcellular localization of CDC25B by binding to a specific site and that 14-3-3σ has effects on CDC25B other than the control of its subcellular localization.

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