Advances and perspectives of the architecture of hemidesmosomes: Lessons from structural biology

Hemidesmosomes (HD) are adhesive protein complexes that mediate stable attachment of basal epithelial cells to the underlying basement membrane. The organization of HDs relies on a complex network of protein-protein interactions, in which integrin α6β4 and plectin play an essential role. Here we summarize the current knowledge of the structure of hemidesmosomal proteins, which includes the structures of the first and second fibronectin type III (FnIII) domains and the calx-β domain of the integrin β4 subunit, the actin binding domain of plectin, and two non-overlapping pairs of spectrin repeats of plectin and BPAG1e. Binding of plectin to the β4 subunit is critical for the formation and the stability of HDs. The recent 3D structure of the primary complex between the integrin β4 subunit and plectin has provided a first insight into the macromolecular recognition mechanisms responsible for HD assembly. Two missense mutations in β4 linked to non lethal forms of epidermolysis bullosa map on the plectin-binding surface. Finally, the formation of the β4-plectin complex induces conformational changes in β4 and plectin, suggesting that their interaction may be subject to allosteric regulation.     

Cryo-electron microscopy for structural biology:current status and future perspectives

Recently, significant technical breakthroughs in both hardware equipment and software algorithms have enabled cryo-electron microscopy(cryo-EM) to become one of the most important techniques in biological structural analysis. The technical aspects of cryo-EM define its unique advantages and the direction of development. As a rapidly emerging field, cryo-EM has benefitted from highly interdisciplinary research efforts. Here we review the current status of cryo-EM in the context of structural biology and discuss the technical challenges. It may eventually merge structural and cell biology at multiple scales.     

Evolutionary cell biology: Lessons from diversity

Novel perspectives emerge from a recent conference on the origins of eukaryotic cells, which covered phylogenetics, population genetics and evolutionary consequences of energy requirements and host–pathogen interactions.     

Enterococcus infection biology: Lessons from invertebrate host models

The enterococci are commensals of the gastrointestinal tract of many metazoans, from insects to humans. While they normally do not cause disease in the intestine, they can become pathogenic when they infect sites outside of the gut. Recently, the enterococci have become important nosocomial pathogens, with the majority of human enterococcal infections caused by two species, Enterococcus faecalis and Enterococcus faecium. Studies using invertebrate infection models have revealed insights into the biology of enterococcal infections, as well as general principles underlying host innate immune defense. This review highlights recent findings on Enterococcus infection biology from two invertebrate infection models, the greater wax moth Galleria mellonella and the free-living bacteriovorous nematode Caenorhabditis elegans.     

Lessons from crystals grown in the Advanced Protein Crystallisation Facility for conventional crystallisation applied to structural biology

The crystallographic quality of protein crystals that were grown in microgravity has been compared to that of crystals that were grown in parallel on earth gravity under otherwise identical conditions. A goal of this comparison was to assess if a more accurate 3D-structure can be derived from crystallographic analysis of the former crystals. Therefore, the properties of crystals prepared with the Advanced Protein Crystallisation Facility (APCF) on earth and in orbit during the last decade were evaluated. A statistical analysis reveals that about half of the crystals produced under microgravity had a superior X-ray diffraction limit with respect of terrestrial controls. Eleven protein structures could be determined at previously unachieved resolutions using crystals obtained in the APCF. Microgravity induced features of the most relevant structures are reported. A second goal of this study was to identify the cause of the crystal quality enhancement useful for structure determination. No correlations between the effect of microgravity and other system-dependent parameters, such as isoelectric point or crystal solvent content, were found except the reduced convection during the crystallisation process. Thus, crystal growth under diffusive regime appears to be the key parameter explaining the beneficial effect of microgravity on crystal quality. The mimicry of these effects on earth in gels or in capillary tubes is discussed and the practical consequences for structural biology highlighted.     

When cells lose water: Lessons from biophysics and molecular biology

Organisms living in deserts and anhydrobiotic species are useful models for unraveling mechanisms used to overcome water loss. In this context, late embryogenesis abundant (LEA) proteins and sugars have been extensively studied for protection against desiccation stress and desiccation tolerance. This article aims to reappraise the current understanding of these molecules by focusing on converging contributions from biochemistry, molecular biology, and the use of biophysical tools. Such tools have greatly advanced the field by uncovering intriguing aspects of protein 3-D structure, such as folding upon stress. We summarize the current research on cellular responses against water deficit at the molecular level, considering both plausible water loss-sensing mechanisms and genes governing signal transduction pathways. Finally, we propose models that could guide future experimentation, for example, by concentrating on the behavior of selected proteins in living cells.     

Opportunities for yeast metabolic engineering: Lessons from synthetic biology

Constant progress in genetic engineering has given rise to a number of promising areas of research that facilitated the expansion of industrial biotechnology. The field of metabolic engineering, which utilizes genetic tools to manipulate microbial metabolism to enhance the production of compounds of interest, has had a particularly strong impact by providing new platforms for chemical production. Recent developments in synthetic biology promise to expand the metabolic engineering toolbox further by creating novel biological components for pathway design. The present review addresses some of the recent advances in synthetic biology and how these have the potential to affect metabolic engineering in the yeast Saccharomyces cerevisiae. While S. cerevisiae for years has been a robust industrial organism and the target of multiple metabolic engineering trials, its potential for synthetic biology has remained relatively unexplored and further research in this field could strongly contribute to industrial biotechnology. This review also addresses are general considerations for pathway design, ranging from individual components to regulatory systems, overall pathway considerations and whole-organism engineering, with an emphasis on potential contributions of synthetic biology to these areas. Some examples of applications for yeast synthetic biology and metabolic engineering are also discussed.     

Theoretical molecular biology: prospectives and perspectives

I briefly discuss some aspects of theoretical molecular biology. Specifically, I include the issues of searches for homologies via string matchings, for patterns of specific nucleotide groupings and of sequence-structure relationship. The various approaches developed in order to achieve this end are described, attempting to convey some of the excitement in this quickly growing field.     

BioMEMS and cellular biology: perspectives and applications

The ability to culture cells has revolutionized hypothesis testing in basic cell and molecular biology research. It has become a standard methodology in drug screening, toxicology, and clinical assays, and is increasingly used in regenerative medicine. However, the traditional cell culture methodology essentially consisting of the immersion of a large population of cells in a homogeneous fluid medium and on a homogeneous flat substrate has become increasingly limiting both from a fundamental and practical perspective. Microfabrication technologies have enabled researchers to design, with micrometer control, the biochemical composition and topology of the substrate, and the medium composition, as well as the neighboring cell type in the surrounding cellular microenvironment. Additionally, microtechnology is conceptually well-suited for the development of fast, low-cost in vitro systems that allow for high-throughput culturing and analysis of cells under large numbers of conditions. In this interview, Albert Folch explains these limitations, how they can be overcome with soft lithography and microfluidics, and describes some relevant examples of research in his lab and future directions.     

Timeline: the march of structural biology

One hundred years ago, we knew very little about biological macromolecules and had no tools available to study their structure. Structural biology is now a mature science. New structures are being solved at an ever-increasing rate and there are important new initiatives to determine all the protein folds that are used by biological systems (structural genomics). This article traces some of the key developments in the field.     

Nuclear architecture: the cell biology of a laminopathy

Lamin mutations cause muscular dystrophies, but the mechanism is unclear. A?new study shows that lamin mutant worms display muscle-specific defects linked to altered subnuclear localization of heterochromatin, leading to altered gene expression.