Acidic residues of yeast frataxin have an essential role in Fe-S cluster assembly

Friedreich ataxia is caused by decreased levels of frataxin, a mitochondrial acidic protein that is assumed to act as chaperone in the assembly of Fe-S clusters on the scaffold Isu protein. Frataxin has the in vitro capacity to form iron-loaded multimers, which also suggests an iron storage function. It has been reported that alanine substitution of residues in an acidic ridge of yeast frataxin (Yfh1) elicits loss of iron binding in vitro but has no effect on Fe-S cluster synthesis in vivo. Here, we show that a marked change in the electrostatic properties of a specific region of Yfh1 surface - by substituting two or four acidic residues by lysine or alanine, respectively - impairs Fe-S cluster assembly, weakens the interaction between Yfh1 and Isu1, and increases oxidative damage. Therefore, the acidic ridge is essential for the Yfh1 function and is likely to be involved in iron-mediated protein-protein interactions.     

Developmental profile of the aberrant dopamine D2 receptor response in striatal cholinergic interneurons in DYT1 dystonia

Background

DYT1 dystonia, a severe form of genetically determined human dystonia, exhibits reduced penetrance among carriers and begins usually during adolescence. The reasons for such age dependence and variability remain unclear.

Methods and Results

We characterized the alterations in D2 dopamine receptor (D2R) signalling in striatal cholinergic interneurons at different ages in mice overexpressing human mutant torsinA (hMT). An abnormal excitatory response to the D2R agonist quinpirole was recorded at postnatal day 14, consisting of a membrane depolarization coupled to an increase in spiking frequency, and persisted unchanged at 3 and 9 months in hMT mice, compared to mice expressing wild-type human torsinA and non-transgenic mice. This response was blocked by the D2R antagonist sulpiride and depended upon G-proteins, as it was prevented by intrapipette GDP-β-S. Patch-clamp recordings from dissociated interneurons revealed a significant increase in the Cav2.2-mediated current fraction at all ages examined. Consistently, chelation of intracellular calcium abolished the paradoxical response to quinpirole. Finally, no gross morphological changes were observed during development.

Conclusions

These results suggest that an imbalanced striatal dopaminergic/cholinergic signaling occurs early in DYT1 dystonia and persists along development, representing a susceptibility factor for symptom generation.     

Binding properties of the calcium-activated F2 isoform of Lethocerus troponin C

While in most muscles contraction is triggered by calcium effluxes, insect flight muscles are also activated by mechanical stretch. We are interested in understanding the role that the troponin C protein, usually the calcium sensor, plays in stretch activation. In the flight muscles of Lethocerus, a giant water bug often used as a model system, there are two isoforms of TnC, F1 and F2, present in an approximately 10:1 ratio. F1 TnC is responsible for activating the muscle following a stretch, whereas F2 TnC produces a sustained contraction, the magnitude of which depends on the concentration of Ca(2+) in the fiber. We have previously shown that F1 TnC binds only one Ca(2+) ion in its C-terminal domain and that interaction with troponin H, the insect ortholog of troponin I, is insensitive to Ca(2+). Here, we have studied the effect of Ca(2+) and Mg(2+) on the affinities of the interaction of F2 TnC with troponin H peptides. We show that the presence of two Ca(2+) ions, one in each of the globular domains, increases the affinity for TnH by at least 1 order of magnitude. The N lobe has a lower affinity for Ca(2+), but it is also sensitive to Mg(2+). The C lobe is insensitive to Mg(2+) as previously demonstrated by mutations of the individual EF-hands. The interaction with TnH seems also to have significant structural differences from that observed for the F1 TnC isoform. We discuss how our findings could account for stretch activation.     

Pollen emission from olive trees and concentrations of airborne pollen in an urban area of North Sardinia

In this study the seasonal and daily variations in olive airborne pollen concentrations were measured in the atmosphere of Sassari (Italy) and the olive pollen emission was monitored in the countryside during the flowering period in 1995 and 1996, in order to detect the patterns of change in the atmosphere. The intensity and the timing of pollination was also studied in relation to phenological stages occurrence. In addition, the influence of the main meteorological parameters on pollen emission and airborne pollen dispersal in the city was assessed. Airborne pollen reached its highest concentration a few days before the peak of pollen emission in 1995 but several days after it in 1996 (6 days). Analysis of hourly concentrations shows that the maximum emission and dispersion recorded during the observation period occurred in the middle of the day. Significant regressions were found between hourly temperature and air humidity values and hourly pollen concentrations recorded in the olive grove for almost every day studied, indicating a negative correlation between humidity and pollen concentration and a positive correlation between pollen concentration and temperature. On the other hand, no significant correlation was observed between the meteorological parameters and pollen concentration recorded in the urban area.     

Identification of an evolutionarily conserved family of inorganic polyphosphate endopolyphosphatases

Inorganic polyphosphate (poly-P) consists of just a chain of phosphate groups linked by high energy bonds. It is found in every organism and is implicated in a wide variety of cellular processes (e.g. phosphate storage, blood coagulation, and pathogenicity). Its metabolism has been studied mainly in bacteria while remaining largely uncharacterized in eukaryotes. It has recently been suggested that poly-P metabolism is connected to that of highly phosphorylated inositol species (inositol pyrophosphates). Inositol pyrophosphates are molecules in which phosphate groups outnumber carbon atoms. Like poly-P they contain high energy bonds and play important roles in cell signaling. Here, we show that budding yeast mutants unable to produce inositol pyrophosphates have undetectable levels of poly-P. Our results suggest a prominent metabolic parallel between these two highly phosphorylated molecules. More importantly, we demonstrate that DDP1, encoding diadenosine and diphosphoinositol phosphohydrolase, possesses a robust poly-P endopolyphosphohydrolase activity. In addition, we prove that this is an evolutionarily conserved feature because mammalian Nudix hydrolase family members, the three Ddp1 homologues in human cells (DIPP1, DIPP2, and DIPP3), are also capable of degrading poly-P.     

Preparation and configuration of racemic and optically active analgesic cycloaminoalkylnaphthalenes

Cycloaminoalkylnaphthalene 3 shows interesting opioid‐like analgesic properties. It possesses two chiral centers and can exist as two racemic pairs and four diastereomers. Since the binding of opioids with receptors is stereoselective, it was important to have the two racemic pairs as well as the four diastereomers. In this paper the synthesis of the (2R,3S/2S,3R) racemate and the (2R,3S) and (2S,3R) enantiomers of the 1,2‐dimethyl‐3‐[2‐(6‐hydroxynaphthyl)]‐3‐hydroxypyrrolidine 3 is considered and the determination of absolute configuration is described. The (2R,3S/2S,3R)‐ 3 racemate and the (2R,3S)‐ 3 and (2S,3R)‐ 3 enantiomers were prepared by reaction of the racemic and optically active 1,2‐dimethyl‐3‐pyrrolidone 2, respectively, with the lithiation product obtained from 2‐bromo‐6‐tetrahydropyranyloxy‐naphthalene 1 and acidic hydrolysis. The above‐mentioned enantiomers of 3 were also obtained by optical resolution via fractional crystallization of the salts with d ‐ and l ‐tartaric acids. The configuration of the optically active compounds was determined by X‐ray analysis of a crystal of (−)‐(2S,3R)‐ 3 · HCl · H₂O. The pharmacological test HPT showed that (−)‐(2S,3R)‐ 3 · HCl · H₂O enantiomer is able to induce opioid‐like analgesia with a relative potency 1.5 times that of (2R,3S/2S,3R)‐ 3 and ∼1.5 times that of morphine. Chirality 11:21–28, 1999. © 1999 Wiley‐Liss, Inc.     

Elemental variability in the coralline alga Lithophyllum yemenense as an archive of past climate in the Gulf of Aden (NW Indian Ocean)

This study presents the first algal thallus (skeleton) archive of Asian monsoon strength and Red Sea influence in the Gulf of Aden. Mg/Ca, Li/Ca, and Ba/Ca were measured in Lithophyllum yemenense from Balhaf (Gulf of Aden) using laser ablation inductively coupled plasma mass spectrometry, and Mg/Ca ratio oscillation was used to reconstruct the chronology (34 y). Oscillations of element rates corresponding to the algal growth between 1974 and 2008 were compared with recorded climate and oceanographic variability. During this period, sea surface temperatures (SST) in Balhaf recorded a warming trend of 0.55°C, corresponding to an increase in Mg and Li content in the algal thallus of 2.1 mol‐% and 1.87 μmol‐%, respectively. Lithophyllum yemenense recorded decadal SST variability by Li/Ca, and the influence of the Pacific El‐Niño Southern Oscillation on the NW Indian Ocean climate system by Ba/Ca. Additionally, algal Mg/Ca, Li/Ca, and Ba/Ca showed strong and significant correlations with All Indian Rainfall in the decadal range indicating that these proxies can be useful for tracking variability in the Indian monsoon system, possibly due to changes of the surface wind system, with deep water upwelling in summer, and a distinct seasonality.