Activities and response to DNA damage of latent and active sequence-specific DNA binding forms of mouse p53

The mouse p53 protein generated by alternative splicing (p53as) has amino acid substitutions at its C terminus that result in constitutively active sequence-specific DNA binding (active form), whereas p53 protein itself binds inefficiently (latent form) unless activated by C-terminal modification. Exogenous p53as expression activated transcription of reporter plasmids containing p53 binding sequences and inhibited growth of mouse and human cells lacking functional endogenous p53. Inducible p53as in stably transfected p53 null fibroblasts increased p21WAF1/Cip-1/Sdi and decreased bcl-2 protein steady-state levels. Endogenous p53as and p53 proteins differed in response to cellular DNA damage. p53 protein was induced transiently in normal keratinocytes and fibroblasts whereas p53as protein accumulation was sustained in parallel with induction of p21WAF1/Cip-1/Sdi protein and mRNA, in support of p53as transcriptional activity. Endogenous p53 and p53as proteins in epidermal tumor cells responded to DNA damage with different kinetics of nuclear accumulation and efficiencies of binding to a p53 consensus DNA sequence. A model is proposed in which C-terminally distinct p53 protein forms specialize in functions, with latent p53 forms primarily for rapid non-sequence-specific binding to sites of DNA damage and active p53 forms for sustained regulation of transcription and growth.

Biosynthesis of the Monoterpenes Limonene and Carvone in the Fruit of Caraway1 : I. Demonstration of Enzyme Activities and Their Changes with Development

The biosynthesis of the monoterpenes limonene and carvone in the fruit of caraway (Carum carvi L.) proceeds from geranyl diphosphate via a three-step pathway. First, geranyl diphosphate is cyclized to (+)-limonene by a monoterpene synthase. Second, this intermediate is stored in the essential oil ducts without further metabolism or is converted by limonene-6-hydroxylase to (+)-trans-carveol. Third, (+)-trans-carveol is oxidized by a dehydrogenase to (+)-carvone. To investigate the regulation of monoterpene formation in caraway, we measured the time course of limonene and carvone accumulation during fruit development and compared it with monoterpene biosynthesis from [U-14C]Suc and the changes in the activities of the three enzymes. The activities of the enzymes explain the profiles of monoterpene accumulation quite well, with limonene-6-hydroxylase playing a pivotal role in controlling the nature of the end product. In the youngest stages, when limonene-6-hydroxylase is undetectable, only limonene was accumulating in appreciable levels. The appearance of limonene-6-hydroxylase correlates closely with the onset of carvone accumulation. At later stages of fruit development, the activities of all three enzymes declined to low levels. Although this correlates closely with a decrease in monoterpene accumulation, the latter may also be the result of competition with other pathways for substrate.

Expertise Development in Volleyball: the Role of Early Sport Activities and Players’ Age and Height

The purpose of this study was to analyse the developmental pathway of skilled and less skilled volleyball players by focusing on the quantity and type of sporting activities, as well as their age and height in comparison to peers in those experiences. Retrospective interviews were conducted to provide a longitudinal and detailed account of sport involvement of 30 skilled and 30 less skilled volleyball players (15 male and 15 female players per group) throughout different developmental stages (stage 1: 8-12 years; stage 2: 13-16 years; stage 3: 17-20 years). Results indicated that the developmental pathway of these volleyball players (i.e. skilled and less skilled) was characterized by an early diversified sport involvement with a greater participation in sport activities during stages 1 and 2. However, skilled players specialized later in volleyball (between age 14 and 15) and performed more hours of volleyball at stage 3 (from 17 years of age onwards). Also, skilled players (male and female) were younger in both the diversified sport activities and volleyball at the later stages of development (i.e. stages 2 and 3), and skilled female players were taller than peers in those activities in the early stages of development (i.e. stages 1 and 2). The present findings suggest early diversification as a feasible pathway to reach expertise in volleyball and highlight the importance of practicing with older peers once specialization in the main sport has occurred. The findings highlight the need for coaches and sport programs to consider different stimuli existing within the training environment (i.e. characteristics of athletes, such as age and height) that influence the quality of practice and contribute to players’ expertise development.