Revealing the dynamic mechanisms by which protein complexes cooperate to establish cell polarity
Lead Research Organisation:
Newcastle University
Department Name: Biosciences Institute
Abstract
Most metazoan cells are polarised; they show structural asymmetries essential to their function. Polarisation is driven by effectors, notably PAR proteins, that must be efficient at both (1) sensing polarity-cues in order to become asymmetrically localised, and (2) signalling to a variety of downstream targets that execute the cell polarisation programme. How can PARs accommodate these functional requirements polarising cells as diverse as neurons, epithelia and asymmetrically dividing cells? Recent work from our lab has shed light onto this longstanding-question by identifying distinct PAR-complexes that specialise in either sensing the polarity cue or in transducing the signal. We propose that cooperation between specialised PAR-complexes can confer plasticity to the PAR network (Rodriguez_et_al.,Dev.Cell.2017).
This project will elucidate the dynamics and molecular basis of the cooperation between the identified PAR-complexes in the genetically tractable C.elegans embryo. The project will benefit from our combined approach of unbiased screens and sophisticated phenotypic analyses of polarity conditional mutants, which is advancing our understanding of PAR regulation and function (Sundar_et_al.,Elife2018 -under revision, minor corrections-; Rodriguez_et_al.,Dev. Cell.2017; Rodriguez_et_al.,Nat.Cell.Biol.2013). Experiments will tackle the following aims:
[1] Determine in vivo how the dynamic cooperation of PAR-complexes is achieved
Our data indicates that this PAR-complex cooperation depends on a dynamic exchange of the key polarity-signalling component, aPKC between the different PAR-complexes, ensuring an asymmetric domain of active PARs. Using optimized reporter strains for super-resolution imaging the student will test this hypothesis by characterizing aPKC dynamics in vivo at a nano-scale resolution.
[2] Identify regulators of the PAR-complex cooperation
We have developed an innovative genetic screen and we will perform proteomics analyses (supported by a recently awarded BBSRC new investigator grant, May 2018), identifying candidate regulators of PAR-complex cooperation. The student will join our screening efforts and will characterise 2-3 promising candidates using the imaging tools/techniques developed in aim1.
[3] In silico analyses of PAR dynamic cooperation
The student will help model the functional interactions identified in aims1&2 to uncover biological mechanisms and aid experimental design.
By elucidating the fundamental principles governing cell polarity and tissue homeostasis, we will provide new avenues for drug design strategies, particularly in developmental diseases including cancer (Martin-Belmonte_et_al., Nat.Rev.Cancer.2018) or in complex processes where cell polarity is perturbed, such as aging (Budovsky_et_al.,Biogerontology,2011; Florian_et_al.,CellStemCell,2012).
Moreover the supervisory teams expertise in cell polarity, super-resolution imaging and genetic-screens (JR-Newcastle), quantitative and automated images analyses (RB-Newcastle) and cell polarity modelling (NS-Liverpool) will afford the student an ideal interdisciplinary research-training environment.
This project will elucidate the dynamics and molecular basis of the cooperation between the identified PAR-complexes in the genetically tractable C.elegans embryo. The project will benefit from our combined approach of unbiased screens and sophisticated phenotypic analyses of polarity conditional mutants, which is advancing our understanding of PAR regulation and function (Sundar_et_al.,Elife2018 -under revision, minor corrections-; Rodriguez_et_al.,Dev. Cell.2017; Rodriguez_et_al.,Nat.Cell.Biol.2013). Experiments will tackle the following aims:
[1] Determine in vivo how the dynamic cooperation of PAR-complexes is achieved
Our data indicates that this PAR-complex cooperation depends on a dynamic exchange of the key polarity-signalling component, aPKC between the different PAR-complexes, ensuring an asymmetric domain of active PARs. Using optimized reporter strains for super-resolution imaging the student will test this hypothesis by characterizing aPKC dynamics in vivo at a nano-scale resolution.
[2] Identify regulators of the PAR-complex cooperation
We have developed an innovative genetic screen and we will perform proteomics analyses (supported by a recently awarded BBSRC new investigator grant, May 2018), identifying candidate regulators of PAR-complex cooperation. The student will join our screening efforts and will characterise 2-3 promising candidates using the imaging tools/techniques developed in aim1.
[3] In silico analyses of PAR dynamic cooperation
The student will help model the functional interactions identified in aims1&2 to uncover biological mechanisms and aid experimental design.
By elucidating the fundamental principles governing cell polarity and tissue homeostasis, we will provide new avenues for drug design strategies, particularly in developmental diseases including cancer (Martin-Belmonte_et_al., Nat.Rev.Cancer.2018) or in complex processes where cell polarity is perturbed, such as aging (Budovsky_et_al.,Biogerontology,2011; Florian_et_al.,CellStemCell,2012).
Moreover the supervisory teams expertise in cell polarity, super-resolution imaging and genetic-screens (JR-Newcastle), quantitative and automated images analyses (RB-Newcastle) and cell polarity modelling (NS-Liverpool) will afford the student an ideal interdisciplinary research-training environment.
Organisations
People |
ORCID iD |
| Josana Rodriguez (Primary Supervisor) | |
| Iolo Squires (Student) |