Ribosome biogenesis in eukaryotes is a major cellular activity mobilizing the products of over 200 transcriptionally co-regulated genes referred to as the rRNA and ribosome biosynthesis (RRB) regulon. We investigated the function of an essential, uncharacterized gene of this regulon, renamed RRP36. We show that the Rrp36p protein is nucleolar and interacts with 90S and pre-40S pre-ribosomal particles. Its depletion affects early cleavages of the 35S pre-rRNA and results in a rapid decrease in mature 18S rRNA levels. Rrp36p is a novel component of the 90S pre-ribosome, the assembly of which has been suggested to result from the stepwise incorporation of several modules, including the tUTP/UTP-A, PWP2/UTP-B and UTP-C sub-complexes. We show that Rrp36p depletion does not impair the incorporation of these sub-complexes and the U3 snoRNP into pre-ribosomes. In contrast, depletion of components of the UTP-A or UTP-B modules, but not Rrp5p, prevents Rrp36p recruitment and reduces its accumulation levels. In parallel, we studied the human orthologue of Rrp36p in HeLa cells and we show that the function of this protein in early cleavages of the pre-rRNA has been conserved through evolution in eukaryotes.

Prp43p is a RNA helicase required for pre-mRNA splicing and for the synthesis of large and small ribosomal subunits. The molecular functions and modes of regulation of Prp43p during ribosome biogenesis remain unknown. We demonstrate that the G-patch protein Pfa1p, a component of pre-40S pre-ribosomal particles, directly interacts with Prp43p. We also show that lack of Gno1p, another G-patch protein associated with Prp43p, specifically reduces Pfa1p accumulation, whereas it increases the levels of the pre-40S pre-ribosomal particle component Ltv1p. Moreover, cells lacking Pfa1p and depleted for Ltv1p show strong 20S pre-rRNA accumulation in the cytoplasm and reduced levels of 18S rRNA. Finally, we demonstrate that Pfa1p stimulates the ATPase and helicase activities of Prp43p. Truncated Pfa1p variants unable to fully stimulate the activity of Prp43p fail to complement the 20S pre-rRNA processing defect of Deltapfa1 cells depleted for Ltv1p. Our results strongly suggest that stimulation of ATPase/helicase activities of Prp43p by Pfa1p is required for efficient 20S pre-rRNA-to-18S rRNA conversion.

BACKGROUND: Although it is a crucial cellular process required for both normal development and to face stress conditions, the control of programmed cell death in plants is not fully understood. We previously reported the isolation of ATXR5 and ATXR6, two PCNA-binding proteins that could be involved in the regulation of cell cycle or cell death. A yeast two-hybrid screen using ATXR5 as bait captured AtIPS1, an enzyme which catalyses the committed step of myo-inositol (MI) biosynthesis. atips1 mutants form spontaneous lesions on leaves, raising the possibility that MI metabolism may play a role in the control of PCD in plants. In this work, we have characterised atips1 mutants to gain insight regarding the role of MI in PCD regulation. METHODOLOGY/PRINCIPAL FINDINGS:- lesion formation in atips1 mutants depends of light intensity, is due to PCD as evidenced by TUNEL labelling of nuclei, and is regulated by phytohormones such as salicylic acid - MI and galactinol are the only metabolites whose accumulation is significantly reduced in the mutant, and supplementation of the mutant with these compounds is sufficient to prevent PCD - the transcriptome profile of the mutant is extremely similar to that of lesion mimic mutants such as cpr5, or wild-type plants infected with pathogens. CONCLUSION/SIGNIFICANCE: Taken together, our results provide strong evidence for the role of MI or MI derivatives in the regulation of PCD. Interestingly, there are three isoforms of IPS in Arabidopsis, but AtIPS1 is the only one harbouring a nuclear localisation sequence, suggesting that nuclear pools of MI may play a specific role in PCD regulation and opening new research prospects regarding the role of MI in the prevention of tumorigenesis. Nevertheless, the significance of the interaction between AtIPS1 and ATXR5 remains to be established.

PURPOSE: Ultrasound (US) has emerged in the field of regional anaesthesia in the past few years, as it allows physicians to simultaneously see the needle, the targeted nerves, and the vessels to avoid. Nevertheless, anatomical knowledge is essential for identifying all of the structures seen on the US screen. US also allows an in vivo approach to the variations of nerves and vessels. The aim of this study was to describe the anatomical structures of the axilla through a dissection, an anatomical section and US images performed during daily regional anaesthesia. This work will also discuss the usefulness of US in studying anatomical variations of vasculonervous structures. METHODS: The axillary region of an embalmed adult cadaver was dissected in the department of Anatomy, and anatomical sections of another embalmed and frozen cadaver were also performed. During the same period, fortuitous anatomical variations discovered during daily routine axillary US-guided nerve blocks were recorded in the department of Anaesthesiology. RESULTS: The anatomical dissection and sections allowed correlations to be made and structures to be identified on the US images. The most frequent anatomical variations found were double axillary artery, numerous axillary veins, variant location of the musculocutaneous nerve and posterior location of the brachial plexus in relation to the axillary artery. CONCLUSION: Anatomical knowledge is of major importance for US-guided regional anaesthesia. US scan offers a new approach to anatomical variations of the vasculonervous bundle at the junction of the axilla and the upper arm.

Summary Introduction: Bone turnover decreases from adolescence into adulthood, but does not reach a nadir until the fourth decade. Biochemical markers of bone turnover reflect different processes before and after peak bone mass, so hormonal influences on bone turnover may differ before and after peak bone mass. Objectives: To describe the changes in bone turnover and hormones relevant to bone metabolism from adolescence into adulthood, and to identify which hormones correlate with bone turnover before and after peak bone mass. Design/participants: Two measurements of bone turnover markers and hormones were obtained five to nine years apart in 116 healthy males and females recruited from secondary schools and general practices. Correlations were examined cross-sectionally and longitudinally. Results: DHEAS correlated negatively with bone turnover cross-sectionally and longitudinally (r -0.59 to -0.69) in males and females under age 25. IGF-1 correlated positively with aminoterminal propeptide of type I procollagen (PINP) cross-sectionally and longitudinally (r 0.35) in women over age 25. After correction for change in BMI there were significant longitudinal correlations between DHEAS and bone turnover in women under 25 (r -0.62,-0.66) and IGF-1 and PINP in women over 25 (r 0.56). Conclusions: We have described changes in bone turnover and hormones from adolescence into adulthood. DHEAS correlates with bone turnover before peak bone mass which may represent a direct effect on bone metabolism or the role of DHEAS as a substrate for conversion to other sex steroids. IGF-1 is correlated with PINP in women after peak bone mass which may reflect an influence on cortical modelling.

OBJECTIVE: A minority of patients with ankylosing spondylitis (AS) fail to respond to infliximab treatment. This study compared the circulating infliximab concentration and the presence of clinical symptoms in patients continuously treated with infliximab or after treatment interruption. METHODS: Patients with active AS were randomly assigned at week 0 to receive infliximab either at weeks 4, 6, 10, and then every 6 weeks (continuous treatment), or at weeks 4, 6, and 10 and then upon symptom recurrence (on-demand treatment). The circulating concentration of infliximab was determined early during treatment and at weeks 46 and 52 for the continuous treatment group or upon relapse for the on-demand group. Response in the continuous treatment group was defined at week 58 using the ASsessment in AS International Working Group Criteria for 20% improvement. RESULTS: Among the 93 patients in the continuous treatment group, treatment failure was not associated with a low circulating concentration of infliximab, either during early treatment or at 1 year. Eleven (39.2%) of the 28 nonresponders had an infliximab concentration of >10 mug/ml at week 52, whereas 9 (13.8%) of the 65 responders had an infliximab concentration of <1 mug/ml. In the on-demand group, the infliximab concentration at relapse closely correlated with the time to relapse. However, 24 (36.9%) of 65 patients had a resurgence of clinical symptoms at an infliximab concentration of >10 mug/ml, whereas 25 patients (38.4%) had a relapse at an infliximab concentration of <0.5 mug/ml. CONCLUSION: Responsiveness to infliximab treatment is highly heterogeneous among individuals with AS, and this parameter overcomes the circulating infliximab concentration to explain treatment success or failure.

The H/ACA RNAs represent an abundant, evolutionarily conserved and functionally diverse class of non-coding RNAs. Many H/ACA RNAs direct pseudouridylation of rRNAs and snRNAs, while members of the rapidly growing group of 'orphan' H/ACA RNAs participate in pre-rRNA processing, telomere synthesis and probably, in other nuclear processes. The yeast snR30 'orphan' H/ACA snoRNA has long been known to function in the nucleolytic processing of 18S rRNA, but its molecular role remained unknown. Here, we provide biochemical and genetic evidence demonstrating that during pre-rRNA processing, two evolutionarily conserved sequence elements in the 3'-hairpin of snR30 base-pair with short pre-rRNA sequences located in the eukaryote-specific internal region of 18S rRNA. The newly discovered snR30-18S base-pairing interactions are essential for 18S rRNA production and they constitute a complex snoRNA target RNA transient structure that is novel to H/ACA RNAs. We also demonstrate that besides the 18S recognition motifs, the distal part of the 3'-hairpin of snR30 contains an additional snoRNA element that is essential for 18S rRNA processing and that functions most likely as a snoRNP protein-binding site.

In Schizosaccharomyces pombe, septum formation is intricately controlled by proteins which constitute the SIN (Septum Initiation Network) signalling cascade. The SIN ensures the coordination between mitotic exit and cytokinesis. Yeast spg1p is a core component of the SIN pathway and we have previously characterized the two orthologs of this G-protein in Arabidopsis thaliana (named AtSGP1 and 2). In this work, the cell and tissue expression of AtSGP genes during plant development has been analysed using AtSGP promoter::GUS fusions in stably transformed A. thaliana lines. AtSGP1 promoter activity was restricted to the quiescent centre, collumella cells, stomata guard cells and the stele while AtSGP2 promoter activity was detected in atrichoblasts, trichomes and pollen. The observed promoter activities are in accordance with publicly available pollen, stomata guard cell and root transcriptome data. Two-hybrid experiments previously evidenced an interaction between AtMAP3Kepsilon1 and AtSGP1. The AtMAP3Kepsilon1 promoter activity was detected in root apices, trichomes and ovule integuments. A genetic approach involving both markers of these specialized cells and mutant backgrounds was used to reinforce our hypothesis. It appears that, although highly conserved between plants and fungi, the spg1p G-protein has evolved in plants to perform a function different from the SIN pathway. Interestingly, cells expressing AtSGPs possessed limited or null mitotic activity. Our data suggests that AtSGP are crucial signalling components involved either in early cell fate specification, or in the final steps of cell differentiation. This is an interesting starting point for a wider study devoted to functional experiments designed to test these hypotheses.

ABSTRACT: BACKGROUND: Histone acetyltransferases (HATs) play critical roles in the regulation of chromatin structure and gene expression. Arabidopsis genome contains 12 HAT genes, but the biological functions of many of them are still unknown. In this work, we studied the evolutionary relationship and cellular functions of the two Arabidopsis HAT genes homologous to the MYST family members. RESULTS: An extensive phylogenetic analysis of 105 MYST proteins revealed that they can be divided into 5 classes, each of which contains a specific combination of protein modules. The two Arabidopsis MYST proteins, HAM1 and HAM2, belong to a "green clade", clearly separated from other families of HATs. Using a reverse genetic approach, we show that HAM1 and HAM2 are a functionally redundant pair of genes, as single Arabidopsis ham1 and ham2 mutants displayed a wild-type phenotype, while no double mutant seedling could be recovered. Genetic analysis and cytological study revealed that ham1ham2 double mutation induced severe defects in the formation of male and female gametophyte, resulting in an arrest of mitotic cell cycle at early stages of gametogenesis. RT-PCR experiments and the analysis of transgenic plants expressing the GUS reporter gene under the HAM1 or the HAM2 promoter showed that both genes displayed an overlapping expression pattern, mainly in growing organs such as shoots and flower buds. CONCLUSION: The work presented here reveals novel properties for MYST HATs in Arabidopsis. In addition to providing an evolutionary relationship of this large protein family, we show the evidence of a link between MYST and gamete formation as previously suggested in mammalian cells. A possible function of the Arabidopsis MYST protein-mediated histone acetylation during cell division is suggested.

Peak bone mass is an important determinant of bone mass in later life, but the age of peak bone mass is still unclear. We found that bone size and density increase and bone turnover decreases until age 25. It may be possible to influence bone accrual into the third decade. INTRODUCTION: Peak bone mass is a major determinant of bone mass in later life. Bone growth and maturation is site-specific, and the age of peak bone mass is still unclear. It is important to know the age to which bone accrual continues so strategies to maximise bone mass can be targeted appropriately. This study aims to ascertain the age of lumbar spine peak bone mass. METHODS: We measured lumbar spine BMC, estimated volume and BMAD by DXA and biochemical markers of bone turnover in 116 healthy males and females ages 11 to 40, followed up at an interval of five to nine years. RESULTS: The majority of peak bone mass was attained by the mid-twenties. Increases in BMC in adolescents and young adults were mostly due to increases in bone size. Bone turnover markers decreased through adolescence and the third decade and the decreasing rate of change in bone turnover corresponded with the decreasing rate of change in lumbar spine measurements. CONCLUSIONS: Skeletal maturation and bone mineral accrual at the lumbar spine continues into the third decade.