Synthetic In Vivo Reconstitution of Asymmetric Cell Division
type: CreativeWork
description: Asymmetric cell division (ACD) is the process whereby one dividing cell gives rise to two daughter cells of differing fates. ACD is known to be controlled, at least in part, by protein complexes that reside asymmetrically at the cortex of dividing cells. These asymmetric domains are involved in orienting the division plane, localising proteins important for cell fate, as well as for generating polarity in cytosolic structures important for the dispatch of cell fate determinants. One protein complex of particular interest is the Par complex, which is known to asymmetrically localise to the plasma membrane in many systems. It is known that an asymmetric Par complex is required (in many systems), for orienting the division plane, and for generating asymmetry in the density of microtubules in the anaphase-B central spindle (at least in Drosophila). However, whether the Par complex is sufficient to generate these features, or whether they rely on other polarity pathways, is unknown. In this work, I sought to address this question, by developing two techniques for reimparting cortical polarity back into unpolarised cells (cells that have lost polarity). The first of these techniques relies on protein micropatterning, which is the process by which proteins can be ‘printed’ onto a surface. For successful establishment of this assay, a new system of micropatterning, based on ‘anchoring’ proteins to Fibrinogen (a large glycoprotein complex), was developed. Fibrinogen-anchoring remarkably improves the micropatterning of previously difficult-to-pattern proteins, and gives a universal patterning platform upon which complex, multi-protein patterning experiments can be performed. For the first time, protein micropatterns were used to control the subcellular localisation of cell receptors. Further, with this technology, intracellular plasma membrane asymmetries, akin to those that occur in multicellular organisms, could be reconstituted. The second technology for reconstituting pla…
abstract: Asymmetric cell division (ACD) is the process whereby one dividing cell gives rise to two daughter cells of differing fates. ACD is known to be controlled, at least in part, by protein complexes that reside asymmetrically at the cortex of dividing cells. These asymmetric domains are involved in orienting the division plane, localising proteins important for cell fate, as well as for generating polarity in cytosolic structures important for the dispatch of cell fate determinants. One protein complex of particular interest is the Par complex, which is known to asymmetrically localise to the plasma membrane in many systems. It is known that an asymmetric Par complex is required (in many systems), for orienting the division plane, and for generating asymmetry in the density of microtubules in the anaphase-B central spindle (at least in Drosophila). However, whether the Par complex is sufficient to generate these features, or whether they rely on other polarity pathways, is unknown. In this work, I sought to address this question, by developing two techniques for reimparting cortical polarity back into unpolarised cells (cells that have lost polarity). The first of these techniques relies on protein micropatterning, which is the process by which proteins can be ‘printed’ onto a surface. For successful establishment of this assay, a new system of micropatterning, based on ‘anchoring’ proteins to Fibrinogen (a large glycoprotein complex), was developed. Fibrinogen-anchoring remarkably improves the micropatterning of previously difficult-to-pattern proteins, and gives a universal patterning platform upon which complex, multi-protein patterning experiments can be performed. For the first time, protein micropatterns were used to control the subcellular localisation of cell receptors. Further, with this technology, intracellular plasma membrane asymmetries, akin to those that occur in multicellular organisms, could be reconstituted. The second technology for reconstituting pla…
author: Watson, Joseph
language: eng
keywords: Mitosis, Asymmetric Cell Division, Reconstitution, Protein Design, Micropatterning
license: https://apollo8-f-pro.lib.cam.ac.uk/bitstreams/4d5b5646-4e1e-42f4-9a4d-ccfcb6824e4f/download
license: https://www.rioxx.net/licenses/all-rights-reserved/
date published: 2020-08-28
date modified: 2026-07-22T22:24:23Z
provider: Cambridge University
publisher: Networked Digital Library of Theses and Dissertations
type: WebPage
source organization: University of Cambridge

PropertyValue
property ID: DOI
value: 10.17863/CAM.63175