The establishment of cell polarity is a vital requirement for cellular processes such as proper cell division, growth and movement. Cell polarization relies on different internal and external cues in order to reorient the cell growth machinery along the axis of polarity. The core mechanisms involved in establishment of polarized growth are highly conserved from yeast to humans. Cells of the fission yeast Schizosaccharomyces pombe grow in a highly polarized fashion, with cell growth restricted to the cell tips, making fission yeast an excellent model system to study polarized growth. Here I describe a system for long-term live-cell imaging of fission yeast polarized growth that is stress free, physiological and accessible to media change and drug addition. I use this improved imaging system along with yeast genetics and drug perturbations to address how cell polarity is established and maintained in fission yeast. I have shown that fission yeast growth polarity depends on competition and cooperation among three distinct internal polarity cues: 1) A microtubule-based cue involving Tea1/Tea4 polarity proteins positively regulates polarized growth, initially at the “old” cell end (i.e., the end that pre-existed in the mother cell) and later at the “new” cell end (i.e. the end that is generated by septation), in order to initiate the transition from monopolar to bipolar growth (also known as New End Take-Off, or “NETO”). 2) An actin cable-based cue “clears” polarity proteins from the new end immediately after cytokinesis thereby reinforcing old-end growth. As a result perturbation of actin cable-based transport by either deleting actin cable nucleator For3 or cable-based transporter Myo52 results in premature bipolar growth. 3) A novel “memory-based” growth polarity cue helps to establish polarized growth in the absence of the microtubule-based cue. This memory-based cue is dependent on the predicted transmembrane proteins Rax1/Rax2. In the absence of both Tea1/Tea4 cue and Rax1/Rax2 cue, cells depend on septation cue and grow exclusively from the cell ends generated by septation. Furthermore, both Tea1/Tea4 and Rax1/Rax2 cue are important to maintain polarized growth under various environmental stresses. In fission yeast, during interphase, nucleus is positioned at the centre of the cell and this precise positioning of nucleus, which is important for defining the position of cytokinetic ring is thought to be exclusively MT-dependent. Here I show that MT-independent nuclear movement exists in fission yeast and this nuclear movement is mediated by actin cables and type myosin myo52. Furthermore, I show that actin cable might be important for buffering the pushing forces generated by MTs on the nucleus. In this way both microtubules and actin cables are involved in nuclear movement in fission yeast.
| Identifer | oai:union.ndltd.org:bl.uk/oai:ethos.bl.uk:735730 |
| Date | January 2017 |
| Creators | Ashraf, Sanju |
| Contributors | Sawin, Ken ; Goryachev, Andrew |
| Publisher | University of Edinburgh |
| Source Sets | Ethos UK |
| Detected Language | English |
| Type | Electronic Thesis or Dissertation |
| Source | http://hdl.handle.net/1842/28728 |
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