Understanding the structure and function of ASPM, a protein essential for normal mitosis.

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Background: Abnormal spindle-like, microcephaly-associated (ASPM) protein is essential for normal mitosis. Mutations in ASPM affect the plane of cell division resulting in the disorder primary autosomal recessive microcephaly (MCPH), which is characterized by a small brain and cognitive impairment. Its predicted the N-terminal microtubule binding domain and tandem pair of calponin homology domains probably localize it to mitotic spindle poles. The structure of its large central block of 81 'IQ' (isoleucine-glutamine) motifs, which probably bind calmodulin, is unknown, and could be long and linear, or compact depending on how calmodulin interacts with the IQ motifs. The C-terminal region contains binding sites for further interacting proteins, the identity of which are only just being discovered (Bond Lab). Objectives: Our main objective is to understand the mechanism by which ASPM functions in mitosis. Our first aim is to determine the structure of ASPM, to enable us to determine how it might be organized within the spindle pole. Our second aim is to characterize its interactions with its binding partners in vitro, and use this information to interrogate how these interactions contribute to its function in vivo. Novelty & Timeliness: This new collaboration brings together complimentary expertise from the two supervisors in cell and molecular/structural biology, and provides a novel opportunity to investigate the structure and function of ASPM in cell division, one of the key biological processes.Experimental approach: The project will exploit the ability of the Peckham lab to express and purify domains from ASPM to study their structure and function using a range of techniques from protein expression and purification, biochemical assays, high content imaging, electron microscopy, crystallization, and super-resolution microscopy approaches combined with the strong background, tools, techniques and novel reagents utilised to study ASPM in cells from the Bond Lab.