Dephosphorylation of human cyclin-dependent kinases by protein phosphatase type 2Cα and β2 isoforms
(2000) In Journal of Biological Chemistry 275(44). p.34744-34749- Abstract
We previously reported that the activating phosphorylation on cyclin-dependent kinases in yeast (Cdc28p) and in humans (Cdk2) is removed by type 2C protein phosphatases. In this study, we characterize this PP2C-like activity ha HeLa cell extract and determine that it is due to PP2Cβ2, a novel PP2Cβ isoform, and to PP2Cα. PP2Cα and PP2Cβ2 co-purified with Mg2+-dependent Cdk2/Cdk6 phosphatase activity in DEAE-Sepharose, Superdex-200, and Mono Q chromatographies. Moreover, purified recombinant PP2Cα and PP2Cβ2 proteins efficiently dephosphorylated monomeric Cdk2/Cdk6 in vitro. The dephosphorylation of Cdk2 and Cdk6 by PP2C isoforms was inhibited by the binding of cyclins. We found that the PP2C-like activity in HeLa cell... (More)
We previously reported that the activating phosphorylation on cyclin-dependent kinases in yeast (Cdc28p) and in humans (Cdk2) is removed by type 2C protein phosphatases. In this study, we characterize this PP2C-like activity ha HeLa cell extract and determine that it is due to PP2Cβ2, a novel PP2Cβ isoform, and to PP2Cα. PP2Cα and PP2Cβ2 co-purified with Mg2+-dependent Cdk2/Cdk6 phosphatase activity in DEAE-Sepharose, Superdex-200, and Mono Q chromatographies. Moreover, purified recombinant PP2Cα and PP2Cβ2 proteins efficiently dephosphorylated monomeric Cdk2/Cdk6 in vitro. The dephosphorylation of Cdk2 and Cdk6 by PP2C isoforms was inhibited by the binding of cyclins. We found that the PP2C-like activity in HeLa cell extract, partially purified HeLa PP2Cα and PP2Cβ2 isoforms, and the recombinant PP2Cs exhibited a comparable substrate preference for a phosphothreonine containing substrate, consistent with the conservation of threonine residues at the site of activating phosphorylation in CDKs.
(Less)
- author
- Cheng, Aiyang ; Kaldis, Philipp LU and Solomon, Mark J.
- publishing date
- 2000-11-03
- type
- Contribution to journal
- publication status
- published
- in
- Journal of Biological Chemistry
- volume
- 275
- issue
- 44
- pages
- 34744 - 34749
- publisher
- American Society for Biochemistry and Molecular Biology
- external identifiers
-
- scopus:0034602268
- pmid:10934208
- ISSN
- 0021-9258
- DOI
- 10.1074/jbc.M006210200
- language
- English
- LU publication?
- no
- id
- ddd34f4c-ecb3-4bfe-96d4-3eb5b7515870
- date added to LUP
- 2019-09-18 14:31:47
- date last changed
- 2024-05-29 00:59:03
@article{ddd34f4c-ecb3-4bfe-96d4-3eb5b7515870, abstract = {{<p>We previously reported that the activating phosphorylation on cyclin-dependent kinases in yeast (Cdc28p) and in humans (Cdk2) is removed by type 2C protein phosphatases. In this study, we characterize this PP2C-like activity ha HeLa cell extract and determine that it is due to PP2Cβ2, a novel PP2Cβ isoform, and to PP2Cα. PP2Cα and PP2Cβ2 co-purified with Mg<sup>2+</sup>-dependent Cdk2/Cdk6 phosphatase activity in DEAE-Sepharose, Superdex-200, and Mono Q chromatographies. Moreover, purified recombinant PP2Cα and PP2Cβ2 proteins efficiently dephosphorylated monomeric Cdk2/Cdk6 in vitro. The dephosphorylation of Cdk2 and Cdk6 by PP2C isoforms was inhibited by the binding of cyclins. We found that the PP2C-like activity in HeLa cell extract, partially purified HeLa PP2Cα and PP2Cβ2 isoforms, and the recombinant PP2Cs exhibited a comparable substrate preference for a phosphothreonine containing substrate, consistent with the conservation of threonine residues at the site of activating phosphorylation in CDKs.</p>}}, author = {{Cheng, Aiyang and Kaldis, Philipp and Solomon, Mark J.}}, issn = {{0021-9258}}, language = {{eng}}, month = {{11}}, number = {{44}}, pages = {{34744--34749}}, publisher = {{American Society for Biochemistry and Molecular Biology}}, series = {{Journal of Biological Chemistry}}, title = {{Dephosphorylation of human cyclin-dependent kinases by protein phosphatase type 2Cα and β2 isoforms}}, url = {{http://dx.doi.org/10.1074/jbc.M006210200}}, doi = {{10.1074/jbc.M006210200}}, volume = {{275}}, year = {{2000}}, }