Visualizing helicases unwinding DNA at the single molecule level

DNA helicases are motor proteins that catalyze the unwinding of double-stranded DNA into single-stranded DNA using the free energy from ATP hydrolysis. Single molecule approaches enable us to address detailed mechanistic questions about how such enzymes move processively along DNA. Here, an optical method has been developed to follow the unwinding of multiple DNA molecules simultaneously in real time. This was achieved by measuring the accumulation of fluorescent single-stranded DNA-binding protein on the single-stranded DNA product of the helicase, using total internal reflection fluorescence microscopy. By immobilizing either the DNA or helicase, localized increase in fluorescence provides information about the rate of unwinding and the processivity of individual enzymes. In addition, it reveals details of the unwinding process, such as pauses and bursts of activity. The generic and versatile nature of the assay makes it applicable to a variety of DNA helicases and DNA templates. The method is an important addition to the single-molecule toolbox available for studying DNA processing enzymes

Myo1c is designed for the adaptation response in the inner ear

The molecular motor, Myo1c, a member of the myosin family, is widely expressed in vertebrate tissues. Its presence at strategic places in the stereocilia of the hair cells in the inner ear and studies using transgenic mice expressing a mutant Myo1c that can be selectively inhibited implicate it as the mediator of slow adaptation of mechanoelectrical transduction, which is required for balance. Here, we have studied the structural, mechanical and biochemical properties of Myo1c to gain an insight into how this molecular motor works. Our results support a model in which Myo1c possesses a strain-sensing ADP-release mechanism, which allows it to adapt to mechanical load

Occupational image, organisational image and identity in dirty work: Intersections of organisational efforts and media accounts

This article proposes that media representations of an occupational category may intersect with organizations’ efforts to construct a positive organizational identity and image. We fuse three streams of literature namely, organizational identity and image, media and the social construction of reality, and dirty work to extend extant literature on organizational identity and image. Attention is drawn to occupational image as the position of an occupational category in society. We contend that occupational image is likely to influence the decisions and actions taken by organizations and its members, in particular when the occupation is central to the organization’s mission. Occupational image is partly informed by the media. We analyse one year of media coverage of a dirty work occupation, specifically exotic dancing, and identify various ways in which the media portrays the exotic dancing occupation and the organizations providing these services. We focus upon two of these categories, namely Public (dis) Order and Art and Entertainment. We also draw upon a variety of data from one organization, For Your Eyes Only, to explore how organizational efforts to construct a positive organizational identity (based upon professionalism and legitimacy) and image (based upon fantasy, exclusivity and high quality service) intersect these media representations