C-terminal amino acids are essential for human heat shock protein 70 dimerization

The human inducible heat shock protein 70 (hHsp70), which is involved in several major pathologies, including neurodegenerative disorders and cancer, is a key molecular chaperone and contributes to the proper protein folding and maintenance of a large number of protein structures. Despite its role in disease, the current structural knowledge of hHsp70 is almost exclusively based on its Escherichia coli homolog, DnaK, even though these two proteins only share ~50 % amino acid identity. For the first time, we describe a complete heterologous production and purification strategy that allowed us to obtain a large amount of soluble, full-length, and non-tagged hHsp70. The protein displayed both an ATPase and a refolding activity when combined to the human Hsp40. Multi-angle light scattering and bio-layer interferometry analyses demonstrated the ability of hHsp70 to homodimerize. The role of the C-terminal part of hHsp70 was identified and confirmed by a study of a truncated version of hHsp70 that could neither dimerize nor present refolding activity.

Electronic supplementary material

The online version of this article (doi:10.1007/s12192-014-0526-3) contains supplementary material, which is available to authorized users.     

The importomer peroxins are differentially required for peroxisome assembly and meiotic development in Podospora anserina: insights into a new peroxisome import pathway

Peroxisome biogenesis relies on two known peroxisome matrix protein import pathways that are mediated by the receptors PEX5 and PEX7. These pathways converge at the importomer, a peroxisome‐membrane complex that is required for protein translocation into peroxisomes and consists of docking and RING–finger subcomplexes. In the fungus Podospora anserina, the RING–finger peroxins are crucial for meiocyte formation, while PEX5, PEX7 or the docking peroxin PEX14 are not. Here we show that PEX14 and the PEX14‐related protein PEX14/17 are differentially involved in peroxisome import during development. PEX14/17 activity does not compensate for loss of PEX14 function, and elimination of both proteins has no effect on meiocyte differentiation. In contrast, the docking peroxin PEX13, and the peroxins implicated in peroxisome membrane biogenesis PEX3 and PEX19, are required for meiocyte formation. Remarkably, the PTS2 coreceptor PEX20 is also essential for meiocyte differentiation and this function does not require PEX5 or PEX7. This finding suggests that PEX20 can mediate the import receptor activity of specific peroxisome matrix proteins. Our results suggest a new pathway for peroxisome import, which relies on PEX20 as import receptor and which seems critically required for specific developmental processes, like meiocyte differentiation in P. anserina.     

Early specialization for voice and emotion processing in the infant brain

Human voices play a fundamental role in social communication, and areas of the adult "social brain" show specialization for processing voices and their emotional content (superior temporal sulcus, inferior prefrontal cortex, premotor cortical regions, amygdala, and insula). However, it is unclear when this specialization develops. Functional magnetic resonance (fMRI) studies suggest that the infant temporal cortex does not differentiate speech from music or backward speech, but a prior study with functional near-infrared spectroscopy revealed preferential activation for human voices in 7-month-olds, in a more posterior location of the temporal cortex than in adults. However, the brain networks involved in processing nonspeech human vocalizations in early development are still unknown. To address this issue, in the present fMRI study, 3- to 7-month-olds were presented with adult nonspeech vocalizations (emotionally neutral, emotionally positive, and emotionally negative) and nonvocal environmental sounds. Infants displayed significant differential activation in the anterior portion of the temporal cortex, similarly to adults. Moreover, sad vocalizations modulated the activity of brain regions involved in processing affective stimuli such as the orbitofrontal cortex and insula. These results suggest remarkably early functional specialization for processing human voice and negative emotions.     

Evaluation of maximal O₂ uptake with undergraduate students at the University of La Reunion

The maximal rate of O? consumption (VO? max) constitutes one of the oldest fitness indexes established for the measure of cardiorespiratory fitness and aerobic performance. Procedures have been developed in which VO? max is estimated from physiological responses during submaximal exercise. Generally, VO? max is estimated using the classical renowned Astrand-Ryhming test. In young adults, poor fitness and low aerobic performance are often associated with a sedentary lifestyle, which is a well-described factor for the development of obesity and its related disorders such as cardiovascular diseases and type 2 diabetes. In the Indian Ocean, the inhabitants of La Reunion Island, a French overseas department, exhibit an increasing prevalence of obesity and type 2 diabetes. At the University of La Reunion, a new laboratory course involving students was designed to teach the indirect evaluation of their VO? max from the classical Astrand-Ryhming test and using a cycle ergometer as the exercise mode. Inverse and significant correlations were established between the students' fat mass percentages and their VO? max and between their waist-to-hip ratio and VO? max as well. Results from the international physical activity questionnaire showed that most participants in this laboratory were sedentary students. Therefore, this laboratory makes the students practice and understand the use of a classical test to estimate their VO? max. It also alerts them to the correlation between a sedentary lifestyle and higher body fat content. This exercise allowed students to use a scientific method to engage the problem of sedentary lifestyle, which is a real world issue.     

Acquisition and maintenance of resistance to viruses in eukaryotic phytoplankton populations

Viruses are known to play a key role in the regulation of eukaryotic phytoplankton population densities; however, little is known about the mechanisms of how they interact with their hosts and how phytoplankton populations mediate their regulations. Viruses are obligate parasites that depend on host cell machinery for their dissemination in the environment (most of the time through host cell lysis that liberates many new particles). But viruses also depend on a reliable host population to carry on their replication before losing their viability. How do hosts cells survive when they coexist with their viruses? We show that clonal lines of three picoeukaryotic green algae (i.e. Bathycoccus sp., Micromonas sp., Ostreococcus tauri) reproducibly acquire resistance to their specific viruses following a round of infection. Our observations show that two mechanisms of resistance may operate in O. tauri. In the first resistant type, viruses can attach to their host cells but no new particles develop. In the second one, O. tauri acquires tolerance to its virus and releases these viruses consistently. These lines maintained their resistance over a 3‐year period, irrespective of whether or not they were re‐challenged with new viral inoculations. Co‐culturing resistant and susceptible lines revealed resistance to be associated with reduced host fitness in terms of growth rate.     

Communication and data-intensive science in the beginning of the 21st century

The advent of data-intensive science has sharpened our need for better communication within and between the fields of science and technology, to name a few. No one mind can encompass all that is necessary to be successful in controlling and analyzing the data deluge we are experiencing. Therefore, we must bring together diverse fields, communicate clearly, and build crossdisciplinary methods and tools to realize its potential. This article is a summary of the communication issues and challenges as discussed in the Data-Intensive Science (DIS) workshop in Seattle, September 19-20, 2010.     

A yeast-based screen reveals that sulfasalazine inhibits tetrahydrobiopterin biosynthesis

We introduce an approach for detection of drug-protein interactions that combines a new yeast three-hybrid screening for identification of interactions with affinity chromatography for their unambiguous validation. We applied the methodology to the profiling of clinically approved drugs, resulting in the identification of previously known and unknown drug-protein interactions. In particular, we were able to identify off-targets for erlotinib and atorvastatin, as well as an enzyme target for the anti-inflammatory drug sulfasalazine. We demonstrate that sulfasalazine and its metabolites, sulfapyridine and mesalamine, are inhibitors of the enzyme catalyzing the final step in the biosynthesis of the cofactor tetrahydrobiopterin. The interference with tetrahydrobiopterin metabolism provides an explanation for some of the beneficial and deleterious properties of sulfasalazine and furthermore suggests new and improved therapies for the drug. This work thus establishes a powerful approach for drug profiling and provides new insights in the mechanism of action of clinically approved drugs.