ALIX in ciliogenesis
The primary cilium is a solitary organelle consisting of a microtubule-filled core called the axoneme which extends from the basal body derived from centrioles (Figure 1). The primary cilia protrude from the cell surface like antennae in non-dividing cells. Extensive research over the last 50 years has contributed to unravel that primary cilia are involved in diverse processes like cell signalling and cell cycle progression. Since cancer cells typically show uncontrolled cell proliferation and dysregulated intracellular signalling, primary cilia are suggested to be implicated in cancer development. However, the exact mechanisms are yet to be elucidated.
Figure 1- Confocal micrograph shows a primary cilium where the axoneme is stained with anti-acetylated tubulin (green) and DNA is stained with Hoechst (blue). Interestingly, ALIX (red) localizes to the basal body of the primary cilium.
Goal of the project
According to the current need to further characterize how primary cilia influence cancer development at the molecular level the current study aims to elucidate the role of the multifunctional adaptor protein ALIX in primary cilia-function (i.e ciliogenesis). We have preliminary results indicating that ALIX localizes to the basal body of primary cilia (Figure 1) but it remains to characterize how ALIX regulates the assembly or disassembly of primary cilia and/or the cell signaling mediated from this signaling hub. We have previously shown that ALIX plays a crucial role in both early and late steps of cell division and it will therefore be particularly interesting to elucidate if ALIX-regulated ciliogenesis influences cell cycle progression.
In order to address the outlined scientific questions, the candidate will learn and perform a variety of biochemical and imaging-based methods like:
-genetically manipulate expression levels of ALIX and other relevant proteins in cultured cells by RNA interference, ectopic overexpression and CRISPR/Cas9 technology
-biochemical assays like Western blotting analyses will be used to assess cellular protein levels
-confocal fluorescence and superresolution imaging will be used in order to investigate the localization of ALIX and signaling molecules in primary cilia
-live cell imaging in combination with biochemical cell proliferation assays will be performed to investigate cell cycle-progression of cells with normal and low levels of ALIX
Supervision and environment
The project will be carried out at Centre for Cancer Cell Reprogramming, a Norwegian Centre of Excellence located at the Department of Molecular Cell Biology, Institute for Cancer Research, Oslo University Hospital. The master student will work side-by-side with the supervisor Dr. Lene Malerød in the lab to ensure adequate progression in the project group of Dr. Kaisa Haglund within the group of Prof. Harald Stenmark. The department is an excellent scientific institution to learn comprehensive basic science and the master student will benefit from collaborating with highly skilled colleagues and having access to state-of the-art technologies held by the department and associated core facilities.