Generation of a Set of Conditional Analog-sensitive Alleles of Essential Protein Kinases in the Fission Yeast Schizosaccharomyces Pombe

Overview

Journal Cell Cycle

Specialty Cell Biology

Date 2011 Oct 28

PMID 22030861

Citations 23

Authors

Lubos Cipak

Chao Zhang

Ines Kovacikova

Cornelia Rumpf

Eva Miadokova

Kevan M Shokat

Juraj Gregan

Affiliations

Soon will be listed here.

Abstract

The genome of the fission yeast Schizosaccharomyces pombe encodes for 17 protein kinases that are essential for viability. Studies of the essential kinases often require the use of mutant strains carrying conditional alleles. To inactivate these kinases conditionally, we applied a recently developed chemical genetic strategy. The mutation of a single residue in the ATP-binding pocket confers sensitivity to small-molecule inhibitors, allowing for specific inactivation of the modified kinase. Using this approach, we constructed conditional analog-sensitive alleles of 13 essential protein kinases in the fission yeast S. pombe.

Citing Articles

Sororin is an evolutionary conserved antagonist of WAPL.

Prusen Mota I, Galova M, Schleiffer A, Nguyen T, Kovacikova I, Farias Saad C Nat Commun. 2024; 15(1):4729.

PMID: 38830897 PMC: 11148194. DOI: 10.1038/s41467-024-49178-0.

Polarity kinases that phosphorylate F-BAR protein Cdc15 have unique localization patterns during cytokinesis and contributions to preventing tip septation in .

Igarashi M, Bhattacharjee R, Willet A, Gould K MicroPubl Biol. 2023; 2023.

PMID: 37746062 PMC: 10517346. DOI: 10.17912/micropub.biology.000965.

Multiple polarity kinases inhibit phase separation of F-BAR protein Cdc15 and antagonize cytokinetic ring assembly in fission yeast.

Bhattacharjee R, Hall A, Mangione M, Igarashi M, Roberts-Galbraith R, Chen J Elife. 2023; 12.

PMID: 36749320 PMC: 9904764. DOI: 10.7554/eLife.83062.

Reactivity-based chemical-genetic study of protein kinases.

Miranda R, Zhang C RSC Med Chem. 2022; 13(7):783-797.

PMID: 35923719 PMC: 9298188. DOI: 10.1039/d1md00389e.

Phosphoproteomics Meets Chemical Genetics: Approaches for Global Mapping and Deciphering the Phosphoproteome.

Jurcik J, Sivakova B, Cipakova I, Selicky T, Stupenova E, Jurcik M Int J Mol Sci. 2020; 21(20).

PMID: 33076458 PMC: 7588962. DOI: 10.3390/ijms21207637.

References

Gregan J, Rabitsch P, Rumpf C, Novatchkova M, Schleiffer A, Nasmyth K . High-throughput knockout screen in fission yeast. Nat Protoc. 2007; 1(5):2457-64. PMC: 2957175. DOI: 10.1038/nprot.2006.385. View

Knight Z, Shokat K . Features of selective kinase inhibitors. Chem Biol. 2005; 12(6):621-37. DOI: 10.1016/j.chembiol.2005.04.011. View

KREBS E . Nobel Lecture. Protein phosphorylation and cellular regulation I. Biosci Rep. 1993; 13(3):127-42. DOI: 10.1007/BF01149958. View

Guiguen A, Soutourina J, Dewez M, Tafforeau L, Dieu M, Raes M . Recruitment of P-TEFb (Cdk9-Pch1) to chromatin by the cap-methyl transferase Pcm1 in fission yeast. EMBO J. 2007; 26(6):1552-9. PMC: 1829387. DOI: 10.1038/sj.emboj.7601627. View

Dischinger S, Krapp A, Xie L, Paulson J, Simanis V . Chemical genetic analysis of the regulatory role of Cdc2p in the S. pombe septation initiation network. J Cell Sci. 2008; 121(Pt 6):843-53. DOI: 10.1242/jcs.021584. View

Rumpf C, Cipak L, Dudas A, Benko Z, Pozgajova M, Riedel C . Casein kinase 1 is required for efficient removal of Rec8 during meiosis I. Cell Cycle. 2010; 9(13):2657-62. PMC: 3083834. DOI: 10.4161/cc.9.13.12146. View

Rumpf C, Cipak L, Schleiffer A, Pidoux A, Mechtler K, Tolic-Norrelykke I . Laser microsurgery provides evidence for merotelic kinetochore attachments in fission yeast cells lacking Pcs1 or Clr4. Cell Cycle. 2010; 9(19):3997-4004. PMC: 2975032. DOI: 10.4161/cc.9.19.13233. View

Viladevall L, St Amour C, Rosebrock A, Schneider S, Zhang C, Allen J . TFIIH and P-TEFb coordinate transcription with capping enzyme recruitment at specific genes in fission yeast. Mol Cell. 2009; 33(6):738-51. PMC: 2693121. DOI: 10.1016/j.molcel.2009.01.029. View

Sabatinos S, Forsburg S . Molecular genetics of Schizosaccharomyces pombe. Methods Enzymol. 2010; 470:759-95. DOI: 10.1016/S0076-6879(10)70032-X. View

10.

Zhang C, Kenski D, Paulson J, Bonshtien A, Sessa G, Cross J . A second-site suppressor strategy for chemical genetic analysis of diverse protein kinases. Nat Methods. 2005; 2(6):435-41. DOI: 10.1038/nmeth764. View

11.

Fischer E . Phosphorylase and the origin of reversible protein phosphorylation. Biol Chem. 2009; 391(2-3):131-137. DOI: 10.1515/bc.2010.011. View

12.

Das M, Wiley D, Chen X, Shah K, Verde F . The conserved NDR kinase Orb6 controls polarized cell growth by spatial regulation of the small GTPase Cdc42. Curr Biol. 2009; 19(15):1314-9. DOI: 10.1016/j.cub.2009.06.057. View

13.

Forsburg S, Rhind N . Basic methods for fission yeast. Yeast. 2006; 23(3):173-83. PMC: 5074380. DOI: 10.1002/yea.1347. View

14.

Mata J, Marguerat S, Bahler J . Post-transcriptional control of gene expression: a genome-wide perspective. Trends Biochem Sci. 2005; 30(9):506-14. DOI: 10.1016/j.tibs.2005.07.005. View

15.

Hauf S, Biswas A, Langegger M, Kawashima S, Tsukahara T, Watanabe Y . Aurora controls sister kinetochore mono-orientation and homolog bi-orientation in meiosis-I. EMBO J. 2007; 26(21):4475-86. PMC: 2034495. DOI: 10.1038/sj.emboj.7601880. View

16.

Koch A, Hauf S . Strategies for the identification of kinase substrates using analog-sensitive kinases. Eur J Cell Biol. 2010; 89(2-3):184-93. DOI: 10.1016/j.ejcb.2009.11.024. View

17.

Nemoto N, Udagawa T, Ohira T, Jiang L, Hirota K, Wilkinson C . The roles of stress-activated Sty1 and Gcn2 kinases and of the protooncoprotein homologue Int6/eIF3e in responses to endogenous oxidative stress during histidine starvation. J Mol Biol. 2010; 404(2):183-201. PMC: 4378542. DOI: 10.1016/j.jmb.2010.09.016. View

18.

Sabatinos S, Forsburg S . Measuring DNA content by flow cytometry in fission yeast. Methods Mol Biol. 2009; 521:449-61. DOI: 10.1007/978-1-60327-815-7_25. View

19.

Lackner D, Bahler J . Translational control of gene expression from transcripts to transcriptomes. Int Rev Cell Mol Biol. 2008; 271:199-251. DOI: 10.1016/S1937-6448(08)01205-7. View

20.

Bishop A, Ubersax J, Petsch D, Matheos D, Gray N, Blethrow J . A chemical switch for inhibitor-sensitive alleles of any protein kinase. Nature. 2000; 407(6802):395-401. DOI: 10.1038/35030148. View