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Turkish Journal of Biology

Author ORCID Identifier

SEVİLAY GİRGİN: 0000-0002-0244-3919

AYŞE ÇAYDAŞI: 0000-0003-2570-1367

DOI

10.55730/1300-0152.2702

Abstract

Background/aimSpindle Pole Bodies (SPB), functional equivalent of centrosomes in yeast, duplicate through generation of a new SPB next to the old one. Yet, SPBs are dynamic structures that can grow and exchange, and mechanisms that regulate SPB size remain largely unknown. This study aims to elucidate the role of Bud14 in SPB size maintenance in Saccharomyces cerevisiae.Materials and methodsWe employed quantitative fluorescence microscopy to assess the relative and absolute amounts of SPB structural proteins at SPBs of wildtype cells and in cells lacking BUD14 (bud14∆). Quantifications were performed using asynchronous cell cultures as well as cultures synchronously progressing through the cell cycle and upon different cell cycle arrests. We also made use of mutants that allow Bud14 separation of function.ResultsOur results indicate that higher levels of SPB inner, outer, and central plaque proteins are present at the SPBs of bud14∆ cells compared to wildtype cells during anaphase as well as during nocodazole induced M-phase arrest. However, during α-factor mediated G1 arrest, inner and outer plaque proteins responded differently to the absence of BUD14. A Bud14 mutant that cannot interact with the Protein Phosphatase 1 (Glc7) phenocopied bud14∆ in terms of SPB-bound levels of the inner plaque protein Spc110, whereas disruption of Bud14-Kel1-Kel2 complex did not alter Spc110 levels at SPBs. In cells synchronously released from α-factor arrest, lack of Bud14-Glc7 caused increase of Spc110 at the SPBs at early stages of the cell cycle.ConclusionWe identified Bud14 as a critical protein for SPB size maintenance. The interaction of Bud14 with Glc7, but not with the Kelch proteins, is indispensable for restricting levels of Spc110 incorporated into the SPBs.

Keywords

Bud14, centrosome, Glc7, Protein Phosphatase 1, Spc110, Spindle Pole Body

First Page

267

Last Page

278

Creative Commons License

Creative Commons Attribution 4.0 International License
This work is licensed under a Creative Commons Attribution 4.0 International License.

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