Browsing by discipline "Allmän mikrobiologi"

Archaea are known to thrive in different kinds of extreme habitats. Halophilic archaea are found in environments where the salt concentration is high, like in salt lakes and solar salterns. The habitats of halophilic archaea have salt concentration varying from higher than sea water to the saturation of salt. In high salinity habitats, the biodiversity is typically low and archaea are dominant microorganisms. The cell density of haloarchaea can be up to 107 cell/ml.When there are no predators for archaea and other halophilic microbes, viruses are thought to be the driving agents for their evolution. To this date, 130 archaeal viruses are described and 90 of them are known infect halophilic archaea. Most of the isolated haloarchaeal viruses are head-tailed. These head-tailed viruses can be divided into three different groups by the properties of the tail structure. Those three types of head-tailed viruses are myo-, sipho- and podoviruses. The infection cycles, receptors or lysis mechanisms used by archaeal viruses are still poorly known. Few studied archaeal viruses use seemingly similar strategies to attach to the surface of the host cell, to penetrate the cell surface and to release new virions, as bacteriophages and viruses of eukaryotic cells do. Since archaeas differ so much from eukaryotic or bacterial cells, their viruses must have developed different strategies to carry out their infection cycle. Several filamentous viruses of archaea are known to attach to the pilus structures of their host cells and some of the enveloped archaeal viruses attach straight to the cell membrane of the host. These strategies are seemingly similar to those used by bacteriophages and viruses of eukaryotic cells. Strikingly different mechanism to release virions has been seen on SIRV-2, virus of a hyperthermophilic archaeon. The SIRV-2 infection induces pyramid shaped extrusions on the surface of the host cells. In the end of the infection cycle these pyramids open and release new virions from the cell. This is the first lysis mechanism described for an archaeal virus. In this work, the infection cycle of a head-tailed virus infecting extremely halophilic Haloarcula vallismortis was studied. The Haloarcula vallismortis tailed virus 1 (HVTV-1) is head-tailed siphovirus with a double stranded DNA genome. In previous studies the infection cycle of HVTV-1 is described to be lytic. The genome sequence, 3D-structure of the capsid and the major capsid protein of HVTV-1 is known. HVTV-1 infection has been seen to induce large, roundish structures on the surface of the infected cell. These structures may have a role in the course of the infection cycle. Several animal viruses are known to cause massive rearrangements in the host cell so that replication proteins, viral genome and the replication agents needed from the host are concentrated in specific locations. These rearrangements facilitates and enhances the viral replication. The roundish structures induced by HVTV-1 might play the same role as the virus factories of animal viruses do or these structures may be involved in the release of new virions in somewhat similar manner as those virus induced pyramid structures. In this study two main goals were set: 1.) To explore the adsorption of HVTV-1 in more details and see if the host cell receptor could be solved. 2.) To solve the possible role of the virus induced structure seen in the host cell and to find out, in which point of the infection cycle those structures appear. Based on this study, it can be said that the adsorption of HVTV-1 is efficient and very fast. The virus attaches to the archaella structures of the host cell and the attachment is mediated by the tail of the virus. One archaella may serve as a receptor for several viruses and they are not saturated. The intracellular phase of the infection cycle of HVTV-1 is long and the virus production starts 8 hours after infection. Virions are released when the host cells is lysed 12 hours after the infection. Two hours before the lysis, two virus induced structural changes of the cell are seen. The roundish structures at the surface of the cell and wide areas in the cytoplasm filled by a structure looking similar as lipid membranes. At the same time when the virus production begins, at least two virus induced or coded proteins are produced in the cell. The other one of these proteins was identified and it is predicted to be the virus encoded ribonucleotide reductase. Ribonucleotide reductase is an enzyme known to catalyse the synthesis of deoxyribonucleic acids from ribonucleic acids. If the ribonucleotide reductase is a part of seen structures, the timing of their appearance and the known function of the enzyme, could suggest those structures to play a role in the maturation or release of the virions. Many questions still remains and there would be lot more details to study in the HVTV-1 infection. Especially interesting would be the identification of the other virus induced protein, purifying those roundish structures and analysing them in more details.