Nek7 conformational flexibility and inhibitor binding probed through protein engineering of the R-spine

Nek7 is a serine/threonine protein kinase required for proper spindle formation and cytokinesis. Elevated Nek7 levels have been observed in several cancers, and inhibition of Nek7 might provide a route to the development of cancer therapeutics. To date no selective and potent Nek7 inhibitors have been identified. Nek7 crystal structures exhibit an improperly formed Regulatory-spine (R-spine), characteristic of an inactive kinase. We reasoned that the preference of Nek7 to crystallize in this inactive conformation might hinder attempts to capture Nek7 in complex with Type I inhibitors. Here we have introduced aromatic residues into the R-spine of Nek7 with the aim to stabilize the active conformation of the kinase through R-spine stacking. The strong R-spine mutant Nek7SRSretained catalytic activity and was crystallized in complex with compound 51, an ATP-competitive inhibitor of Nek2 and Nek7. Subsequently, we obtained the same crystal form for wild-type Nek7WTin apo form and bound to compound 51. The R-spines of the three well-ordered Nek7WTmolecules exhibit variable conformations while the R-spines of the Nek7SRSmolecules all have the same, partially stacked configuration. Compound 51 bound to Nek2 and Nek7 in similar modes, but differences in the precise orientation of a substituent highlights features that could be exploited in designing inhibitors that are selective for particular Nek family members. Although the SRS mutations are not required to obtain a Nek7-inhibitor structure, we conclude that it is a useful strategy for restraining the conformation of a kinase in order to promote crystallogenesis

From "a woman's place is in her union" to "strong unions need women": changing gender discourses, policies and realities in the union movement

This article draws on research that I along with others have conducted in various countries and regions of the world, which highlights the changed and evolving gender context within the union movement. From the late 1970s onwards, the campaign for women’s equality and gender democracy within unions gathered pace under the influence of second wave feminism. New discourses were espoused and new practices adopted towards these goals, and some countries, some unions have achieved greater success than have others. The article considers the contemporary landscape, continuity and change in the global union movement’s unfinished gender democracy project

Life after Burberry: shifting experiences of work and non-work life following redundancy

This article sheds new light on neglected areas of recent ‘work-life’ discussions. Drawing on a study of a largely female workforce made redundant by factory relocation, the majority subsequently finding alternative employment in a variety of work settings, the results illustrate aspects of both positive and negative spillover from work to non-work life. In addition, the findings add to the growing number of studies that highlight the conditions under which part-time working detracts from, rather than contributes to, successful work-life balance. The conclusion discusses the need for a more multi-dimensional approach to work-life issues

Front line or all fronts? Women's trade union activism in retail services

This article draws on data from a case study of a trade union campaign to organize part-time women workers in a large supermarket chain. The data indicate that combining paid work, work in the home and increased trade union participation means that the work of women activists and the resistance they encounter in its execution is broader than the customer/ employee interface, or 'front line', that is the current focus in literature on service-sector work and trade unionism. The findings are used to argue that established feminist literature, in which the location and recipients of women's work are conceptualized as multiple and shifting but inter-related, still provide a useful analytical framework for service-sector work. Therefore an 'all fronts' approach may better describe the lives of part-time women workers and trade unionists in the sector. However it is argued that, far from simply being considered as an added burden, trade union activism was a powerful catalyst for change in the home and work lives of the working-class women in the study. © Blackwell Publishing Ltd 2005