PPX, a Novel Protein Serine/threonine Phosphatase Localized to Centrosomes

Overview

Journal EMBO J

Specialties Biotechnology
Molecular Biology

Date 1993 Mar 1

PMID 8384557

Citations 53

Authors

N D Brewis

A J Street

A R Prescott

P T Cohen

Affiliations

Soon will be listed here.

Abstract

The amino acid sequence of a novel mammalian protein phosphatase, termed PPX (and designated PPP4 in the human genome nomenclature), has been deduced from the cDNA and shown to be 65% identical to PP2A alpha and PP2A beta and 45% identical to PPI isoforms, the predicted molecular mass being 35 kDa. PPX was expressed in the baculovirus system. Its substrate specificity and sensitivity to the inhibitors, okadaic acid and microcystin, were similar (but not identical) to the catalytic subunit of PP2A. However, PPX did not bind the 65 kDa regulatory subunit of PP2A. The intracellular localization of PPX was investigated by immunofluorescence using two different antibodies raised against bacterially expressed PPX and a PPX-specific peptide. These showed that although PPX was distributed throughout the cytoplasm and the nucleus, intense staining occurred at centrosomes. The centrosomal staining was apparent in interphase and at all stages of mitosis, except telophase. In contrast, antibodies directed against bacterially expressed PP2A were not specifically localized to centrosomes. The human autoantibody #5051, which stains the pericentriolar material, colocalizes with PPX antibodies, suggesting that PPX may play a role in microtubule nucleation.

Citing Articles

Nuclear-enriched protein phosphatase 4 ensures outer kinetochore assembly prior to nuclear dissolution.

Rocha H, Simoes P, Budrewicz J, Lara-Gonzalez P, Carvalho A, Dumont J J Cell Biol. 2023; 222(3).

PMID: 36719399 PMC: 9930252. DOI: 10.1083/jcb.202208154.

Targeting protein phosphatases for the treatment of inflammation-related diseases: From signaling to therapy.

Pan J, Zhou L, Zhang C, Xu Q, Sun Y Signal Transduct Target Ther. 2022; 7(1):177.

PMID: 35665742 PMC: 9166240. DOI: 10.1038/s41392-022-01038-3.

Protein phosphatases in TLR signaling.

Seumen C, Grimm T, Hauck C Cell Commun Signal. 2021; 19(1):45.

PMID: 33882943 PMC: 8058998. DOI: 10.1186/s12964-021-00722-1.

Functional roles of protein phosphatase 4 in multiple aspects of cellular physiology: a friend and a foe.

Park J, Lee D BMB Rep. 2020; 53(4):181-190.

PMID: 32192570 PMC: 7196183.

Protein phosphatase 2A regulates the nuclear accumulation of the Arabidopsis bZIP protein VIP1 under hypo-osmotic stress.

Tsugama D, Yoon H, Fujino K, Liu S, Takano T J Exp Bot. 2019; 70(21):6101-6112.

PMID: 31504762 PMC: 6859724. DOI: 10.1093/jxb/erz384.

References

Zhang A, Bai G, Browner M, Lee E . Expression of the catalytic subunit of phosphorylase phosphatase (protein phosphatase-1) in Escherichia coli. J Biol Chem. 1992; 267(3):1484-90. View

Vandre D, Wills V . Inhibition of mitosis by okadaic acid: possible involvement of a protein phosphatase 2A in the transition from metaphase to anaphase. J Cell Sci. 1992; 101 ( Pt 1):79-91. DOI: 10.1242/jcs.101.1.79. View

Laemmli U . Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970; 227(5259):680-5. DOI: 10.1038/227680a0. View

Nissen M, LANGAN T, Reeves R . Phosphorylation by cdc2 kinase modulates DNA binding activity of high mobility group I nonhistone chromatin protein. J Biol Chem. 1991; 266(30):19945-52. View

Sneddon A, Cohen P, Stark M . Saccharomyces cerevisiae protein phosphatase 2A performs an essential cellular function and is encoded by two genes. EMBO J. 1990; 9(13):4339-46. PMC: 552220. DOI: 10.1002/j.1460-2075.1990.tb07883.x. View

Berndt N, Cohen P . Renaturation of protein phosphatase 1 expressed at high levels in insect cells using a baculovirus vector. Eur J Biochem. 1990; 190(2):291-7. DOI: 10.1111/j.1432-1033.1990.tb15575.x. View

Buendia B, Draetta G, Karsenti E . Regulation of the microtubule nucleating activity of centrosomes in Xenopus egg extracts: role of cyclin A-associated protein kinase. J Cell Biol. 1992; 116(6):1431-42. PMC: 2289368. DOI: 10.1083/jcb.116.6.1431. View

MacDougall L, Campbell D, Hubbard M, Cohen P . Partial structure and hormonal regulation of rabbit liver inhibitor-1; distribution of inhibitor-1 and inhibitor-2 in rabbit and rat tissues. Biochim Biophys Acta. 1989; 1010(2):218-26. DOI: 10.1016/0167-4889(89)90164-x. View

Mackintosh C, Beattie K, Klumpp S, Cohen P, Codd G . Cyanobacterial microcystin-LR is a potent and specific inhibitor of protein phosphatases 1 and 2A from both mammals and higher plants. FEBS Lett. 1990; 264(2):187-92. DOI: 10.1016/0014-5793(90)80245-e. View

10.

Cohen P . The structure and regulation of protein phosphatases. Annu Rev Biochem. 1989; 58:453-508. DOI: 10.1146/annurev.bi.58.070189.002321. View

11.

Cohen P, Alemany S, Hemmings B, Resink T, Stralfors P, Tung H . Protein phosphatase-1 and protein phosphatase-2A from rabbit skeletal muscle. Methods Enzymol. 1988; 159:390-408. DOI: 10.1016/0076-6879(88)59039-0. View

12.

Siebert M, Hubble R, Mitchison T, Kirschner M . Centrosome development in early mouse embryos as defined by an autoantibody against pericentriolar material. Cell. 1983; 35(3 Pt 2):621-9. DOI: 10.1016/0092-8674(83)90094-6. View

13.

Cohen P, Foulkes J, Holmes C, Nimmo G, Tonks N . Protein phosphatase inhibitor-1 and inhibitor-2 from rabbit skeletal muscle. Methods Enzymol. 1988; 159:427-37. DOI: 10.1016/0076-6879(88)59042-0. View

14.

Church G, Gilbert W . Genomic sequencing. Proc Natl Acad Sci U S A. 1984; 81(7):1991-5. PMC: 345422. DOI: 10.1073/pnas.81.7.1991. View

15.

Kinoshita N, Ohkura H, Yanagida M . Distinct, essential roles of type 1 and 2A protein phosphatases in the control of the fission yeast cell division cycle. Cell. 1990; 63(2):405-15. DOI: 10.1016/0092-8674(90)90173-c. View

16.

Cohen P, Brewis N, Hughes V, Mann D . Protein serine/threonine phosphatases; an expanding family. FEBS Lett. 1990; 268(2):355-9. DOI: 10.1016/0014-5793(90)81285-v. View

17.

Picard A, Capony J, Brautigan D, Doree M . Involvement of protein phosphatases 1 and 2A in the control of M phase-promoting factor activity in starfish. J Cell Biol. 1989; 109(6 Pt 2):3347-54. PMC: 2115967. DOI: 10.1083/jcb.109.6.3347. View

18.

Lee T, Solomon M, Mumby M, Kirschner M . INH, a negative regulator of MPF, is a form of protein phosphatase 2A. Cell. 1991; 64(2):415-23. DOI: 10.1016/0092-8674(91)90649-j. View

19.

Agostinis P, Goris J, Waelkens E, Pinna L, Marchiori F, MERLEVEDE W . Dephosphorylation of phosphoproteins and synthetic phosphopeptides. Study of the specificity of the polycation-stimulated and MgATP-dependent phosphorylase phosphatases. J Biol Chem. 1987; 262(3):1060-4. View

20.

Axton J, Dombradi V, Cohen P, Glover D . One of the protein phosphatase 1 isoenzymes in Drosophila is essential for mitosis. Cell. 1990; 63(1):33-46. DOI: 10.1016/0092-8674(90)90286-n. View