Localization and functionality of microsporidian iron-sulphur cluster assembly proteins

Microsporidia are highly specialized obligate intracellular parasites of other eukaryotes (including humans) that show extreme reduction at the molecular, cellular and biochemical level. Although microsporidia have long been considered as early branching eukaryotes that lack mitochondria, they have recently been shown to contain a tiny mitochondrial remnant called a mitosome. The function of the mitosome is unknown, because microsporidians lack the genes for canonical mitochondrial functions, such as aerobic respiration and haem biosynthesis. However, microsporidial genomes encode several components of the mitochondrial iron-sulphur (Fe-S) cluster assembly machinery. Here we provide experimental insights into the metabolic function and localization of these proteins. We cloned, functionally characterized and localized homologues of several central mitochondrial Fe-S cluster assembly components for the microsporidians Encephalitozoon cuniculi and Trachipleistophora hominis. Several microsporidial proteins can functionally replace their yeast counterparts in Fe-S protein biogenesis. In E. cuniculi, the iron (frataxin) and sulphur (cysteine desulphurase, Nfs1) donors and the scaffold protein (Isu1) co-localize with mitochondrial Hsp70 to the mitosome, consistent with it being the functional site for Fe-S cluster biosynthesis. In T. hominis, mitochondrial Hsp70 and the essential sulphur donor (Nfs1) are still in the mitosome, but surprisingly the main pools of Isu1 and frataxin are cytosolic, creating a conundrum of how these key components of Fe-S cluster biosynthesis coordinate their function. Together, our studies identify the essential biosynthetic process of Fe-S protein assembly as a key function of microsporidian mitosomes.     

Protein degradation and protection against misfolded or damaged proteins

The ultimate mechanism that cells use to ensure the quality of intracellular proteins is the selective destruction of misfolded or damaged polypeptides. In eukaryotic cells, the large ATP-dependent proteolytic machine, the 26S proteasome, prevents the accumulation of non-functional, potentially toxic proteins. This process is of particular importance in protecting cells against harsh conditions (for example, heat shock or oxidative stress) and in a variety of diseases (for example, cystic fibrosis and the major neurodegenerative diseases). A full understanding of the pathogenesis of the protein-folding diseases will require greater knowledge of how misfolded proteins are recognized and selectively degraded.     

Parallel bacterial evolution within multiple patients identifies candidate pathogenicity genes

Bacterial pathogens evolve during the infection of their human host(1-8), but separating adaptive and neutral mutations remains challenging(9-11). Here we identify bacterial genes under adaptive evolution by tracking recurrent patterns of mutations in the same pathogenic strain during the infection of multiple individuals. We conducted a retrospective study of a Burkholderia dolosa outbreak among subjects with cystic fibrosis, sequencing the genomes of 112 isolates collected from 14 individuals over 16 years. We find that 17 bacterial genes acquired nonsynonymous mutations in multiple individuals, which indicates parallel adaptive evolution. Mutations in these genes affect important pathogenic phenotypes, including antibiotic resistance and bacterial membrane composition and implicate oxygen-dependent regulation as paramount in lung infections. Several genes have not previously been implicated in pathogenesis and may represent new therapeutic targets. The identification of parallel molecular evolution as a pathogen spreads among multiple individuals points to the key selection forces it experiences within human hosts.     

Cellular uptake of amelogenin,and its localization to CD63, and Lamp1-positive vesicles

Proteins of the developing enamel matrix include amelogenin, ameloblastin and enamelin. Of these three proteins amelogenin predominates. Protein-protein interactions are likely to occur at the ameloblast Tomes’ processes between membrane-bound proteins and secreted enamel matrix proteins. Such protein-protein interactions could be associated with cell signaling or endocytosis. CD63 and Lamp1 are ubiquitously expressed, are lysosomal integral membrane proteins, and localize to the plasma membrane. CD63 and Lamp1 interact with amelogenin in vitro. In this study our objective was to study the molecular events of intercellular trafficking of an exogenous source of amelogenin, and related this movement to the spatiotemporal expression of CD63 and Lamp1 using various cell lineages. Exogenously added amelogenin moves rapidly into the cell into established Lamp1-positive vesicles that subsequently localize to the perinuclear region. These data indicate a possible mechanism by which amelogenin, or degraded amelogenin peptides, are removed from the extracellular matrix during enamel formation and maturation. Received 27 September 2006; received after revision 24 November 2006; accepted 5 December 2006     

Late Neoproterozoic to early Cambrian sulphur cycle-An isotopic investigation of sedimentary rocks from the Yangtze Platform

The sulphur cycle responds to changes in seawater chemistry, biological evolution and tectonic activity. We follow an isotopic approach in order to constrain the state of the ocean/atmosphere system during late Neoproterozoic and early Cambrian. For this purpose, a sedimentary succession deposited on the Yangtze Platform, South China, was analysed for its sulphur isotopic composition in different S-bearing phases. Redox changes were defined by the degree of pyritization (DOP) values in order to show variations in the oxygenation of the depositional environment. The sulphur isotopic composition of late Neoproterozoic to early Cambrian seawater sulphate ranges from +30‰ to +35‰ as evident from trace sulphate in unaltered carbonates and phosphorites. The isotopic composition for pyrite and organic sulphur varies between -16‰ and +23‰. The apparent sulphur isotopic fractionation between seawater sulphate and pyrite as well as organically bound sulphur varies between 7‰ and 50‰. This large fractionation, as well as its variability suggests a biological origin for pyrite and organically bound sulphur. The temporal evolution of different geochemical proxy signals is comparable for different successions across the Yangtze Platform.     

Lingually induced inhibition of masseteric motoneurones

Zusammenfassung Elektrische Reizung afferenter Fasern niedriger Reizschwelle im Nervus lingualis induzierte in den motorischen Neuronen des Nervus massetericus: (1) IPSPs von kurzer Latenz mittels einer bisynaptischen Bahn, deren Zwischenneurone im Gebiet der supratrigeminalen Nuclei liegen; (2) eine Erregungsphase, die teilweise von einer früh auftretenden Hemmung verdeckt ist; (3) eine Hemmung von langer Latenz mittels einer polysynaptischen Bahn, die sich kaudalwärts vom motorischen Ursprungskern des Nervus trigeminus erstreckt.

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This research was supported by a grant from the USPHS (No. MH-10083). Dr.Goldberg is a NIDR Postdoctoral Fellow. Dr.Nakamura is a Research Fellow from the Department of Neurophysiology, Institute of Brain Research, University of Tokyo (Japan).