Alexandros Linaroudis

An acidic protein aligns magnetosomes along a filamentous structure in magnetotactic bacteria.

Magnetotactic bacteria are widespread aquatic microorganisms that use unique intracellular organelles to navigate along the Earths magnetic field. These organelles, called magnetosomes, consist of membrane-enclosed magnetite crystals that are thought to help to direct bacterial swimming towards growth-favouring microoxic zones at the bottom of natural waters. Questions in the study of magnetosome formation include understanding the factors governing the size and redox-controlled synthesis of the nano-sized magnetosomes and their assembly into a regular chain in order to achieve the maximum possible magnetic moment, against the physical tendency of magnetosome agglomeration. A deeper understanding of these mechanisms is expected from studying the genes present in the identified chromosomal ‘magnetosome island’, for which the connection with magnetosome synthesis has become evident. Here we use gene deletion in Magnetospirillum gryphiswaldense to show that magnetosome alignment is coupled to the presence of the mamJ gene product. MamJ is an acidic protein associated with a novel filamentous structure, as revealed by fluorescence microscopy and cryo-electron tomography. We suggest a mechanism in which MamJ interacts with the magnetosome surface as well as with a cytoskeleton-like structure. According to our hypothesis, magnetosome architecture represents one of the highest structural levels achieved in prokaryotic cells.

Whole Cell Cryo-Electron Tomography Reveals Distinct Disassembly Intermediates of Vaccinia Virus

At each round of infection, viruses fall apart to release their genome for replication, and then reassemble into stable particles within the same host cell. For most viruses, the structural details that underlie these disassembly and assembly reactions are poorly understood. Cryo-electron tomography (cryo-ET), a unique method to investigate large and asymmetric structures at the near molecular resolution, was previously used to study the complex structure of vaccinia virus (VV). Here we study the disassembly of VV by cryo-ET on intact, rapidly frozen, mammalian cells, infected for up to 60 minutes. Binding to the cell surface induced distinct structural rearrangements of the core, such as a shape change, the rearrangement of its surface spikes and de-condensation of the viral DNA. We propose that the cell surface induced changes, in particular the decondensation of the viral genome, are a prerequisite for the subsequent release of the vaccinia DNA into the cytoplasm, which is followed by its cytoplasmic replication. Generally, this is the first study that employs whole cell cryo-ET to address structural details of pathogen-host cell interaction.

Corrigendum: An acidic protein aligns magnetosomes along a filamentous structure in magnetotactic bacteria

This corrects the article DOI: 10.1038/nature04382

SIBS, a powerful concept for automatic segmentation of electron tomograms

A scaling index based segmentation (SIBS) method is proposed in order to improve visualization and interpretation of data obtained by electron tomography. Based on the interpretation of the scaling index as a measure for dimensionality, the pixels/voxels of an image/volume are subdivided into different categories according to the kind of structure they belong to. Using the weighted scaling index method proposed by Räth1 in conjunction with morphological operators, the approach was adapted to the field of electron microscopy, especially to three-dimensional application as needed by electron tomography. The method turns out to be quite effective for linear structures and membranes. Theory, implementation, parameter settings and results obtained with different kinds of data are presented and discussed.

Morphological filtering based on the Minkowski functionals in 3D for segmentation of macromolecular structures in intact eukaryotic cells depicted by cryo-electron tomography

In this contribution, we propose a novel approach to the segmentation of tomographic image data considering topological properties of binarized image components expressed in terms of the Minkowski Functionals in 3D. Electron tomography is a non-invasive method for three-dimensional (3D) reconstruction of cellular sub-structures from a series of projection images (i.e. from a tilt series) recorded with a transmission electron microscope. Data obtained by electron tomography provide a rich source of quantitative information concerning the structural composition and organization of cellular components. It allows to obtain 3D information on structural cellular arrangements at a significantly higher resolution than any other of the currently available imaging modalities. A major challenge, in this context, is the segmentation of the image data with respect to the identification macro-molecular structures such as the actin-cytoskeleton or cell organelles. We introduce a morphological filtering algorithm based on the Minkowski Functionals in 3D for segmentation of macromolecular structures in intact eukaryotic cells depicted by cryo-electron tomography. In mathematical topology, multi-dimensional convex objects can be characterized with respect to shape, structure, and the connectivity of their components using a set of morphological descriptors known as the Minkowski functionals. In a 3D-Euclidian space, these correspond to volume, surface area, mean integral curvature, and the Euler-Poincare characteristic. The morphological filtering procedure is applied to a 3D image data of an intact, ice-embedded Dictyostelium cell obtained by low dose transmission electron microscopy using a tilt series of -50° to +41.5° with an increment of 1.5°. Our method allows to separate cellular components with predefined textural properties, e.g. filamentary or globular structures, from the image data, which may then be studied and interpreted further.