Evidence of 14-3-3 Proteins Contributing to Kinetochore Integrity and Chromosome Congression During Mitosis

Overview

Journal J Cell Sci

Specialty Cell Biology

Date 2024 Jul 11

PMID 38988319

Authors

Guhan Kaliyaperumal Anbalagan

Prakhar Agarwal

Santanu Kumar Ghosh

Affiliations

Soon will be listed here.

Abstract

The 14-3-3 family of proteins are conserved across eukaryotes and serve myriad important regulatory functions in the cell. Homo- and hetero-dimers of these proteins mainly recognize their ligands via conserved motifs to modulate the localization and functions of those effector ligands. In most of the genetic backgrounds of Saccharomyces cerevisiae, disruption of both 14-3-3 homologs (Bmh1 and Bmh2) are either lethal or cells survive with severe growth defects, including gross chromosomal missegregation and prolonged cell cycle arrest. To elucidate their contributions to chromosome segregation, in this work, we investigated their centromere- and kinetochore-related functions of Bmh1 and Bmh2. Analysis of appropriate deletion mutants shows that Bmh isoforms have cumulative and non-shared isoform-specific contributions in maintaining the proper integrity of the kinetochore ensemble. Consequently, Bmh mutant cells exhibited perturbations in kinetochore-microtubule (KT-MT) dynamics, characterized by kinetochore declustering, mis-localization of kinetochore proteins and Mad2-mediated transient G2/M arrest. These defects also caused an asynchronous chromosome congression in bmh mutants during metaphase. In summary, this report advances the knowledge on contributions of budding yeast 14-3-3 proteins in chromosome segregation by demonstrating their roles in kinetochore integrity and chromosome congression.

References

Sluchanko N, Bustos D . Intrinsic disorder associated with 14-3-3 proteins and their partners. Prog Mol Biol Transl Sci. 2019; 166:19-61. DOI: 10.1016/bs.pmbts.2019.03.007. View

Mayordomo I, Sanz P . The Saccharomyces cerevisiae 14-3-3 protein Bmh2 is required for regulation of the phosphorylation status of Fin1, a novel intermediate filament protein. Biochem J. 2002; 365(Pt 1):51-6. PMC: 1222645. DOI: 10.1042/BJ20020053. View

Roberts R, Mosch H, Fink G . 14-3-3 proteins are essential for RAS/MAPK cascade signaling during pseudohyphal development in S. cerevisiae. Cell. 1997; 89(7):1055-65. DOI: 10.1016/s0092-8674(00)80293-7. View

Pearson C, Yeh E, Gardner M, Odde D, Salmon E, Bloom K . Stable kinetochore-microtubule attachment constrains centromere positioning in metaphase. Curr Biol. 2004; 14(21):1962-7. DOI: 10.1016/j.cub.2004.09.086. View

Wach A, Brachat A, Rebischung C, Philippsen P . Heterologous HIS3 marker and GFP reporter modules for PCR-targeting in Saccharomyces cerevisiae. Yeast. 1997; 13(11):1065-75. DOI: 10.1002/(SICI)1097-0061(19970915)13:11<1065::AID-YEA159>3.0.CO;2-K. View

Kumar R, Dhali S, Srikanth R, Ghosh S, Srivastava S . Comparative proteomics of mitosis and meiosis in Saccharomyces cerevisiae. J Proteomics. 2014; 109:1-15. DOI: 10.1016/j.jprot.2014.06.006. View

Fraschini R, Formenti E, Lucchini G, Piatti S . Budding yeast Bub2 is localized at spindle pole bodies and activates the mitotic checkpoint via a different pathway from Mad2. J Cell Biol. 1999; 145(5):979-91. PMC: 2133126. DOI: 10.1083/jcb.145.5.979. View

Kumar R . An account of fungal 14-3-3 proteins. Eur J Cell Biol. 2017; 96(2):206-217. DOI: 10.1016/j.ejcb.2017.02.006. View

Cheeseman I, Muller-Reichert T, Drubin D, Barnes G . Mitotic spindle integrity and kinetochore function linked by the Duo1p/Dam1p complex. J Cell Biol. 2001; 152(1):197-212. PMC: 2193660. DOI: 10.1083/jcb.152.1.197. View

10.

Guarente L . Synthetic enhancement in gene interaction: a genetic tool come of age. Trends Genet. 1993; 9(10):362-6. DOI: 10.1016/0168-9525(93)90042-g. View

11.

Gelperin D, WEIGLE J, Nelson K, Roseboom P, Irie K, Matsumoto K . 14-3-3 proteins: potential roles in vesicular transport and Ras signaling in Saccharomyces cerevisiae. Proc Natl Acad Sci U S A. 1995; 92(25):11539-43. PMC: 40437. DOI: 10.1073/pnas.92.25.11539. View

12.

Martens G, Piosik P, Danen E . Evolutionary conservation of the 14-3-3 protein. Biochem Biophys Res Commun. 1992; 184(3):1456-9. DOI: 10.1016/s0006-291x(05)80046-4. View

13.

Mehta G, Agarwal M, Ghosh S . Functional characterization of kinetochore protein, Ctf19 in meiosis I: an implication of differential impact of Ctf19 on the assembly of mitotic and meiotic kinetochores in Saccharomyces cerevisiae. Mol Microbiol. 2014; 91(6):1179-99. DOI: 10.1111/mmi.12527. View

14.

Wang C, Skinner C, Easlon E, Lin S . Deleting the 14-3-3 protein Bmh1 extends life span in Saccharomyces cerevisiae by increasing stress response. Genetics. 2009; 183(4):1373-84. PMC: 2787426. DOI: 10.1534/genetics.109.107797. View

15.

Jin F, Liu H, Li P, Yu H, Wang Y . Loss of function of the Cik1/Kar3 motor complex results in chromosomes with syntelic attachment that are sensed by the tension checkpoint. PLoS Genet. 2012; 8(2):e1002492. PMC: 3271067. DOI: 10.1371/journal.pgen.1002492. View

16.

Prajapati H, Agarwal M, Mittal P, Ghosh S . Evidence of Promoting Sister Kinetochore Mono-orientation During Meiosis in Budding Yeast. G3 (Bethesda). 2018; 8(11):3691-3701. PMC: 6222564. DOI: 10.1534/g3.118.200469. View

17.

Lu M, Prehoda K . A NudE/14-3-3 pathway coordinates dynein and the kinesin Khc73 to position the mitotic spindle. Dev Cell. 2013; 26(4):369-80. PMC: 3786870. DOI: 10.1016/j.devcel.2013.07.021. View

18.

Sanyal K, Ghosh S, Sinha P . The MCM16 gene of the yeast Saccharomyces cerevisiae is required for chromosome segregation. Mol Gen Genet. 1998; 260(2-3):242-50. DOI: 10.1007/s004380050892. View

19.

Huang X, Zheng Z, Wu Y, Gao M, Su Z, Huang Y . 14-3-3 Proteins are Potential Regulators of Liquid-Liquid Phase Separation. Cell Biochem Biophys. 2022; 80(2):277-293. PMC: 8830994. DOI: 10.1007/s12013-022-01067-3. View

20.

Mittal P, Ghule K, Trakroo D, Prajapati H, Ghosh S . Meiosis-Specific Functions of Kinesin Motors in Cohesin Removal and Maintenance of Chromosome Integrity in Budding Yeast. Mol Cell Biol. 2020; 40(8). PMC: 7108822. DOI: 10.1128/MCB.00386-19. View