Phosphomimetic Mutation of the N-Terminal Lid of MDM2 Enhances the Polyubiquitination of p53 through Stimulation of E2-Ubiquitin Thioester Hydrolysis.

Mouse double minute 2 (MDM2) has a phosphorylation site within a lid motif at Ser17 whose phosphomimetic mutation to Asp17 stimulates MDM2-mediated polyubiquitination of p53. MDM2 lid deletion, but not Asp17 mutation, induced a blue shift in the λmax of intrinsic fluorescence derived from residues in the central domain including Trp235, Trp303, Trp323, and Trp329. This indicates that the Asp17 mutation does not alter the conformation of MDM2 surrounding the tryptophan residues. In addition, Phe235 mutation enhanced MDM2 binding to p53 but did not stimulate its ubiquitination function, thus uncoupling increases in p53 binding from its E3 ubiquitin ligase function. However, the Asp17mutation inMDM2 stimulated its discharge of the UBCH5a-ubiquitin thioester adduct (UBCH5a is a ubiquitin-conjugating enzyme E2D 1 UBC4/5 homolog yeast). This stimulation of ubiquitin discharge fromE2 was independent of the p53 substrate. There are now four known effects of the Asp17 mutation on MDM2: (i) it alters the conformation of the isolated N-terminus as defined by NMR; (ii) it induces increased thermostability of the isolated N-terminal domain; (iii) it stimulates the allosteric interaction ofMDM2 with the DNA-binding domain of p53; and (iv) it stimulates a novel protein–protein interaction with the E2-ubiquitin complex in the absence of substrate p53 that, in turn, increases hydrolysis of theE2-ubiquitin thioester bond. These data also suggest a new strategy to disrupt MDM2 function by targeting the E2-ubiquitin discharge reaction.

Construct and content validity of the Greek version of the birth satisfaction scale (G-BSS).

Background: ‘Birth Satisfaction’ is a term that encompasses a woman’s evaluation of her birth experience. The term includes factors such as her appraisal of the quality of care she received, a personal assessment of how she coped, and her reconstructions of what happened on that particular day. Her accounts may be accurate or skewed, yet correspond with her reality of how events unfolded. Objective: To evaluate properties of an instrument designed to measure birth satisfaction in a Greek population of postnatal women. Study design: We assessed factor structure, internal consistency, divergent validity and known-groups discriminant validity of the 30-item Greek Birth Satisfaction Scale – Long Form (30-item G-BSS-LF) and its revised version the 10-item Greek-BSS-Revised (10-item-G-BSS-R), using survey data collected in Athens. Participants: A convenience sample of healthy Greek postnatal women (n = 162) aged 22–46 years who had delivered between 34 and 42 weeks’ gestation. Results: The 30-item-G-BSS-LF performed poorly in terms of factor structure. The short-form 10-item-G-BSS-R performed well in terms of measurement replication of the English equivalent version as a multidimensional instrument. The short-form 10-item-G-BSS-R comprises three subscales which measure distinct but correlated domains of: (1) quality of care provision (4 items), (2) women’s personal attributes (2 items), and (3) stress experienced during labour (4 items). Key conclusions: The 10-item-G-BSS-R is a valid and reliable multidimensional psychometric instrument for measuring birth satisfaction in Greek postnatal women.