The Actin Filament-Binding Protein Coronin Regulates Motility in Plasmodium Sporozoites.

Parasites causing malaria need to migrate in order to penetrate tissue barriers and enter host cells. Here we show that the actin filament-binding protein coronin regulates gliding motility in Plasmodium berghei sporozoites, the highly motile forms of a rodent malaria-causing parasite transmitted by mosquitoes. Parasites lacking coronin show motility defects that impair colonization of the mosquito salivary glands but not migration in the skin, yet result in decreased transmission efficiency. In non-motile sporozoites low calcium concentrations mediate actin-independent coronin localization to the periphery. Engagement of extracellular ligands triggers an intracellular calcium release followed by the actin-dependent relocalization of coronin to the rear and initiation of motility. Mutational analysis and imaging suggest that coronin organizes actin filaments for productive motility. Using coronin-mCherry as a marker for the presence of actin filaments we found that protein kinase A contributes to actin filament disassembly. We finally speculate that calcium and cAMP-mediated signaling regulate a switch from rapid parasite motility to host cell invasion by differentially influencing actin dynamics.

Enhanced x-ray emission from nano-particle doped bacteria.

Recently, it has been greatly appreciated that intense light matter interaction is modified due to the nano- and microstructures in the target by--surface plasmons, laser energy localization scattering etc. Extreme laser intensities produce dense plasmas and collective mechanisms generate energetic electrons, ions and hard x-rays. Recently, it is postulated that the anharmonic electron motion, driven by ultrashort, high-intensity laser pulses, provides a universal mechanism for the laser absorption. Here, we provide the first demonstration of anharmonic-resonance-aided high laser-absorption in a biological system. At intensities of ∼ 10¹6⁻¹8 W/cm², 40 fs pulses excite a plasma formed with E. coli bacteria. The density-inhomogeneities due to the micro- and nanostructures in the bacterial target increase anharmonic resonance (AHR) heating and result in a 104-fold enhancement in the hard x-ray yield compared to plain solid targets. These observations lead to novel high-energy x-ray sources that have implications to lithography, imaging and medical applications.

Plasmodium falciparum coronin organizes arrays of parallel actin filaments potentially guiding directional motility in invasive malaria parasites.

BACKGROUND: Gliding motility in Plasmodium parasites, the aetiological agents of malaria disease, is mediated by an actomyosin motor anchored in the outer pellicle of the motile cell. Effective motility is dependent on a parasite myosin motor and turnover of dynamic parasite actin filaments. To date, however, the basis for directional motility is not known. Whilst myosin is very likely orientated as a result of its anchorage within the parasite, how actin filaments are orientated to facilitate directional force generation remains unexplained. In addition, recent evidence has questioned the linkage between actin filaments and secreted surface antigens leaving the way by which motor force is transmitted to the extracellular milieu unknown. Malaria parasites possess a markedly reduced repertoire of actin regulators, among which few are predicted to interact with filamentous (F)-actin directly. One of these, PF3D7_1251200, shows strong homology to the coronin family of actin-filament binding proteins, herein referred to as PfCoronin. METHODS: Here the N terminal beta propeller domain of PfCoronin (PfCor-N) was expressed to assess its ability to bind and bundle pre-formed actin filaments by sedimentation assay, total internal reflection fluorescence (TIRF) microscopy and confocal imaging as well as to explore its ability to bind phospholipids. In parallel a tagged PfCoronin line in Plasmodium falciparum was generated to determine the cellular localization of the protein during asexual parasite development and blood-stage merozoite invasion. RESULTS: A combination of biochemical approaches demonstrated that the N-terminal beta-propeller domain of PfCoronin is capable of binding F-actin and facilitating formation of parallel filament bundles. In parasites, PfCoronin is expressed late in the asexual lifecycle and localizes to the pellicle region of invasive merozoites before and during erythrocyte entry. PfCoronin also associates strongly with membranes within the cell, likely mediated by interactions with phosphatidylinositol-4,5-bisphosphate (PI(4,5)P2) at the plasma membrane. CONCLUSIONS: These data suggest PfCoronin may fulfil a key role as the critical determinant of actin filament organization in the Plasmodium cell. This raises the possibility that macro-molecular organization of actin mediates directional motility in gliding parasites.

A bright point source of ultrashort hard x-ray pulses using biological cells.

We demonstrate that the interaction of intense femtosecond light on a plain solid substrate can be substantially altered by a few micron layer coating of bacterial cells, live or dead. Using E. Coli cells, we show that at an intensity of 10(16)W cm(-2), the bremsstraahlung hard x-ray emission (up to 300 keV), is increased by more than two orders of magnitude as compared to a plain glass slab. Particle-in-cell simulations carried out by modeling the bacterial cells as ellipsoidal particles show that the hot electron generation is indeed enhanced by the presence of microstructures. This new methodology should pave way for using microbiological systems of varied shapes to control intense laser produced plasmas for EUV/x-ray generation.