Molecular mechanism of poly(ADP-ribosyl)ation by PARP1 and identification of lysine residues as ADP-ribose acceptor sites

Poly(ADP-ribose) polymerase 1 (PARP1) synthesizes poly(ADP-ribose) (PAR) using nicotinamide adenine dinucleotide (NAD) as a substrate. Despite intensive research on the cellular functions of PARP1, the molecular mechanism of PAR formation has not been comprehensively understood. In this study, we elucidate the molecular mechanisms of poly(ADP-ribosyl)ation and identify PAR acceptor sites. Generation of different chimera proteins revealed that the amino-terminal domains of PARP1, PARP2 and PARP3 cooperate tightly with their corresponding catalytic domains. The DNA-dependent interaction between the amino-terminal DNA-binding domain and the catalytic domain of PARP1 increased Vmax and decreased the Km for NAD. Furthermore, we show that glutamic acid residues in the auto-modification domain of PARP1 are not required for PAR formation. Instead, we identify individual lysine residues as acceptor sites for ADP-ribosylation. Together, our findings provide novel mechanistic insights into PAR synthesis with significant relevance for the different biological functions of PARP family member

PARP1 ADP-ribosylates lysine residues of the core histone tails

The chromatin-associated enzyme PARP1 has previously been suggested to ADP-ribosylate histones, but the specific ADP-ribose acceptor sites have remained enigmatic. Here, we show that PARP1 covalently ADP-ribosylates the amino-terminal histone tails of all core histones. Using biochemical tools and novel electron transfer dissociation mass spectrometric protocols, we identify for the first time K13 of H2A, K30 of H2B, K27 and K37 of H3, as well as K16 of H4 as ADP-ribose acceptor sites. Multiple explicit water molecular dynamics simulations of the H4 tail peptide into the catalytic cleft of PARP1 indicate that two stable intermolecular salt bridges hold the peptide in an orientation that allows K16 ADP-ribosylation. Consistent with a functional cross-talk between ADP-ribosylation and other histone tail modifications, acetylation of H4K16 inhibits ADP-ribosylation by PARP1. Taken together, our computational and experimental results provide strong evidence that PARP1 modifies important regulatory lysines of the core histone tail

‘It’s like equality now; it’s not as if it’s the old days’: an investigation into gender identity development and football participation of adolescent girls

This article explores the influence participating in football has on the development of adolescent girls’ gender identity, an area which currently lacks academic attention. Data were taken from an ethnographic study with a group of adolescent girls and boys and compared to Jeanes’ research. A social constructionist framework was deployed with links to both critical theory and feminist literature. Qualitative and participatory methods were used to fully engage with the complex issue of gender identity. The girls within this study were aware of the normative gender expectations linked to ‘being a female’ but did not find this restrictive. The girls moved between many changing identities and organised their ‘web of selves’ accordingly. The apparent need to measure success by the parameters of male standards created a barrier to girls’ identity development

Unreliable is Better: Theoretical and Practical Impulses for Performance Management

This review aims to stimulate discussion about a comprehensive understanding of performance evaluation—namely, the taken-forgranted benefit of maximal reliable performance evaluation, where employee performance is evaluated with high levels of reliability (i.e., large samples of performance observations). So far, the management discipline has ignored the evidence-based view that one’s performance is better under unreliable performance evaluation compared to reliable performance evaluation. Drawing on tournament theory, behavioral research, and real-world sports data, we argue that while reliable performance evaluation boosts only superior employees, unreliable performance evaluation boosts all employees. The mechanisms that drive inferior and superior employees to perform better when evaluated unreliably substantiate that psychological insight is essential for efficient performance management. Overall, we complement the predominant thinking of performance management by offering innovative insights and implications that are significant for academics, employees, and employers

Forks in the Road of Men’s Gender Politics: Men’s Rights vs Feminist Allies

How do men respond to feminist movements and to shifts in the gender order?  In this paper, I introduce the concept of historical gender formation to show how shifting social conditions over the past forty years shaped a range of men’s organized responses to feminism. Focusing on the US, I show how progressive men reacted to feminism in the 1970s by forming an internally contradictory ‘men’s liberation’ movement that soon split into opposing anti-feminist and pro-feminist factions. Three large transformations of the 1980s and 1990s – the professional institutionalization of feminism, the rise of a postfeminist sensibility, and shifts in the political economy (especially deindustrialization and the rise of the neoliberal state) – generated new possibilities. I end by pointing to an emergent moderate men’s rights discourse that appeals to a postfeminist sensibility, and to an increasingly diverse base for men’s work to prevent violence against women

An image study of institutional management within the state of Kansas

Call number: LD2668 .T4 1969 M45Master of Scienc

Opportunities for organoids as new models of aging.

The biology of aging is challenging to study, particularly in humans. As a result, model organisms are used to approximate the physiological context of aging in humans. However, the best model organisms remain expensive and time-consuming to use. More importantly, they may not reflect directly on the process of aging in people. Human cell culture provides an alternative, but many functional signs of aging occur at the level of tissues rather than cells and are therefore not readily apparent in traditional cell culture models. Organoids have the potential to effectively balance between the strengths and weaknesses of traditional models of aging. They have sufficient complexity to capture relevant signs of aging at the molecular, cellular, and tissue levels, while presenting an experimentally tractable alternative to animal studies. Organoid systems have been developed to model many human tissues and diseases. Here we provide a perspective on the potential for organoids to serve as models for aging and describe how current organoid techniques could be applied to aging research

A versatile ceramic capillary membrane reactor system for continuous enzyme-catalyzed hydrolysis

As an alternative to classical batch processes, enzyme-catalyzed hydrolysis can also be carried out continuously. To facilitate this, a continuous ceramic capillary membrane reactor system (CCCMRS) was developed which can be operated with various proteolytic enzymes immobilized on the porous ceramic capillary membranes. This system has several advantages over common batch processes regarding stability, reproducibility and controllability and can easily be adapted to optimal reaction conditions and individual preferences. Two exemplary applications utilizing the CCCMRS were carried out and investigated in long-term stability studies. In the first application the continuous enzymatic cleavage of human IgG into the antibody fragments Fab and Fc by immobilized papain was performed. A total volume of 22 mL of 1 mg mL-1 IgG-solution was enzymatically cleaved over a period of 33.3 h. The antibody cleavage products could be detected in an SEC-HPLC over the whole process time thus indicating long-term stability of the continuous hydrolysis process. The second application investigated the continuous digestion of pea and almond protein isolates by immobilized Alcalase resulting in the generation of a large variety of different peptides. This peptide fingerprint remains constant over a long period of time enabling fractionation and thus making the peptides accessible for further bioactivity studies in sufficient quantities. The constant peptide fingerprint could be shown in the RP-HPLC analysis for all 30 samples with a total volume of 29.7 mL collected over a period of 45 h