Assembly Of Cellular And Viral Proteins Into Nucleoprotein Complexes In Adenovirus Type 2 Infected Cells

Adenovirus (Ad) is a widely studied DNA virus, but the nucleoprotein structure of the viral genome in the cell is poorly characterized. Our objective is to study Ad DNA-protein associations and how these affect the viral life cycle. Most of the viral DNA condensing protein, protein VII, is lost within a few hours of infection and this loss is independent of transcription. Cellular histones associate with the viral DNA after removal of protein VII, with a preferential deposition of H3.3. Micrococcal nuclease accessibility assays at 6 hpi showed laddering of the viral DNA, suggesting the genome is wrapped in physiologically spaced nucleosomes. Although viral DNA continues to associate with H3.3 at late times of infection, the overall level of association with histones is greatly reduced. Knockdown of the H3.3 chaperone HIRA had no effect on the viral life cycle suggesting that other H3.3 chaperones are involved. Our studies have begun to elucidate the nucleoprotein structure of Ad DNA in the infected cell nucleus.

This book contemplates the structure, dynamics and physics of virus particles: From the moment they come into existence by self-assembly from viral components produced in the infected cell, through their extracellular stage, until they recognise and infect a new host cell and cease to exist by losing their physical integrity to start a new infectious cycle. (Bio)physical techniques used to study the structure of virus particles and components, and some applications of structure-based studies of viruses are also contemplated. This book is aimed first at M.Sc. students, Ph.D. students and postdoctoral researchers with a university degree in biology, chemistry, physics or related scientific disciplines who share an interest or are actually working on viruses. We have aimed also at providing an updated account of many important concepts, techniques, studies and applications in structural and physical virology for established scientists working on viruses, irrespective of their physical, chemical or biological background and their field of expertise. We have not attempted to provide a collection of for-experts-only reviews focused mainly on the latest research in specific topics; we have not generally assumed that the reader knows all of the jargon and all but the most recent and advanced results in each topic dealt with in this book. In short, we have attempted to write a book basic enough to be useful to M.Sc and Ph.D. students, as well as advanced and current enough to be useful to senior scientists with an interest in Structural and/or Physical Virology.

The structure of the nucleoprotein core of adenovirus has been investigated. Using a combination of biophysical techniques and nuclease digestion I have determined that the adenovirus core, as isolated with pyridine, shares several physical properties with cellular chromatin. The most notable difference is the lack of a nucleosome repeat in the staph nuclease digestion intermediates from adenovirus cores. I have exploited this difference to study the fate of the nucleoprotein structure of adenovirus DNA early during productive infection. When nuclei from cells infected with 32P-labeled adenovirus are digested with staph nuclease the pattern of labeled digestion intermediates more closely resembles the pattern of cellular intermediates. This result indicates that virus core proteins are replaced, early during infection, with cellular histones. The newly assembled adenovirus nucleosomes differ from the bulk of the cellular chromatin by having a shorter repeat (160 vs. 185 base pairs) and more labile linker and core regions. Experiments with inhibitors of cellular DNA synthesis indicate that adenovirus nucleosomes are assembled independently of cellular. DNA synthesis. Using the Southern blotting technique it is shown that newly synthesized adenovirus DNA is not in a nucleosome repeat late in infection. At this time of infection, however, the input adenovirus genomes persist in a nucleosome structure. The results of these experiments are discussed in terms of the template requirements for eucaryotic transcription and replication.

Virus Structure covers the full spectrum of modern structural virology. Its goal is to describe the means for defining moderate to high resolution structures and the basic principles that have emerged from these studies. Among the topics covered are Hybrid Vigor, Structural Folds of Viral Proteins, Virus Particle Dynamics, Viral Gemone Organization, Enveloped Viruses and Large Viruses. Covers viral assembly using heterologous expression systems and cell extracts Discusses molecular mechanisms in bacteriophage T7 procapsid assembly, maturation and DNA containment Includes information on structural studies on antibody/virus complexes

This book presents a detailed overview of the development of new viral vector-based vaccines before discussing two major applications: preventive vaccines for infectious diseases and therapeutic cancer vaccines. Viral vector-based vaccines hold a great potential for development into successful pharmaceutical products and several examples at the advanced pre-clinical or clinical stage are presented. Nevertheless, the most efforts were focused on novel and very innovative technologies for new generation of vector-based vaccines. Furthermore, specific topics such as delivery and adjuvant and protection strategies for cell-mediated-based vaccines are presented. Given its scope, the book is a “must read” for all those involved in vaccine development, both in academia and industrial vaccine development.

"Principles of Molecular Virology, Fourth Edition" provides an essential introduction to modern virology in a clear and concise manner. It is a highly enjoyable and readable text with numerous illustrations that enhance the reader's understanding of important principles. It contains new material on virus structure, virus evolution, zoonoses, bushmeat, SARS and bioterrorism. The standard version includes a CD-ROM with Flash animations, virtual interactive tutorials and experiments, self-assessment questions, useful online resources, along with the glossary, classification of subcellular infectious agents and history of virology.

This book provides up-to-date information on experimental and computational characterization of the structural and functional properties of viral proteins, which are widely involved in regulatory and signaling processes. With chapters by leading research groups, it features current information on the structural and functional roles of intrinsic disorders in viral proteomes. It systematically addresses the measles, HIV, influenza, potato virus, forest virus, bovine virus, hepatitis, and rotavirus as well as viral genomics. After analyzing the unique features of each class of viral proteins, future directions for research and disease management are presented.