Identification of novel ligands of WDR47, using yeast two-hybrid analysis
Date
2009-12
Authors
McGillewie, L.
Journal Title
Journal ISSN
Volume Title
Publisher
Stellenbosch : University of Stellenbosch
Abstract
The mammalian neocortex contributes to the increasing functional complexity of the mammalian brain,
partly because of its striking organisation into distinct neuronal layers. The development of the neocortex
has been well studied because disrupted neurodevelopment results in several human diseases.
The basic principles of neocortical development have been well established for some time; however the
molecular mechanisms have only recently been identified. One major advance in our understanding of
these molecular mechanisms was the discovery of Reelin, an extracellular matrix protein that directs the
migration of neurons to their final positions in the developing neocortex.
Reelin is a large multi-domain protein that exerts its functions by binding to its ligands on the cell surface
and initiating a signal transduction cascade that ultimately results in cytoskeletal rearrangements. Several
investigations have been undertaken to elucidate the functions of each of these domains to gain a better
understanding reelin’s functions.
We have previously identified the WR40 repeat protein 47 (WDR47), a protein of unknown function, as a
novel putative ligand for the N-terminal reeler domain of reelin. To gain better understanding into the
functional significance of this interaction, the present study sought to identify novel WDR47- interacting
proteins. In order to achieve this, a cDNA encoding a polypeptide that contains the two N-terminal
domains of WDR47, i.e. the Lis homology and the C-terminal Lis homology domain (CTLH) was used as
bait in a Y2H screen of a foetal brain cDNA library. Putative WDR47 ligands were subsequently verified
using 3D in vivo co-localisation.
Results of these analyses showed that SCG10, a microtubule destabilizing protein belonging to the
stathmin family of proteins, interacted with the N-terminal of WDR47. The identification of SCG10 as a
novel WDR47 interacting protein not only sheds some light on the role and function of WDR47 but also
aids in a better understanding of the reelin pathway and cortical lamination. Moreover, the data presented
here, may also provide researchers with new avenues of research into molecular mechanisms involved in
neuronal migration disorders.
Description
Thesis (MScMedSc (Biomedical Sciences. Molecular Biology and Human Genetics. Medical Biochemistry))--University of Stellenbosch, 2009.
Keywords
Reelin, WDR47, Neocortical development, Yeast two-hybrid, Dissertations -- Medicine, Theses -- Medicine, Dissertations -- Medical biochemistry, Theses -- Medical biochemistry