The family
Marseilleviridae
, defined as a group of icosahedral double-stranded DNA viruses with particle size of approximately 250 nm and genome size of 350–380 kbp, belongs to the nucleo-cytoplasmic family of large DNA viruses. The family
Marseilleviridae
is currently classified into lineages A–E. In this study, we isolated 12 or 15 new members of the family
Marseilleviridae
from three sampling locations in Japan. Molecular phylogenetic analysis of the MCP genes showed that the new viruses could be further classified into three groups, hokutoviruses, kashiwazakiviruses, and kyotoviruses. Hokutoviruses were closely related to lineage B, kyotoviruses were related to lineage A, and kashiwazakiviruses were also classified into lineage B but a new putative subgroup of lineage B, revealing the diversity of this lineage. Interestingly, more than two viruses with slightly different MCP genes were isolated from a single water sample from a single location, i.e., two hokutoviruses and one kashiwazakivirus were isolated from a small reservoir, five kashiwazakiviruses from the mouth of a river, and five kyotoviruses from fresh water of a river, suggesting that several milliliters of water samples contain several types of giant viruses. Amoeba cells infected with hokutoviruses or kashiwazakiviruses exhibited a “bunch” formation consisting of normal and infected cells similarly to a tupanvirus, whereas cells infected with kyotoviruses or tokyovirus did not. These results suggest the previously unrecognized local diversity of the family
Marseilleviridae
in aquatic environments.
Downregulation of virus receptors on the cell surface is considered to be important in preventing superinfection. HIV-1 encodes multiple gene products, Env, Vpu, and Nef, involved in downregulation of CD4, a major HIV-1 receptor. We found that simultaneous mutations in both vpu and nef severely impaired virus replication. We examined the involvement of CD4 downregulation mediated by Vpu and Nef in the modification of virus infectivity. The mutation in vpu increased CD4 incorporation into virions without affecting the Env content in it, inhibiting the attachment step of virions to the CD4-positive cell surface. Although a single mutation in nef suppresses virus infectivity via a CD4-independent mechanism, it could augment CD4 incorporation in virions in combination with a vpu mutation. These results indicated that CD4 downregulation was necessary for maintenance of Env function in the virion.
Crystalline Si-based nanosheets were successfully synthesized from CaSi2 by a simple soft chemical synthetic method in solution. By immersing CaSi2 powder or CaSi2/Si substrates in an inositol hexakisphosphate (IP6) solution, Ca atoms were extracted from the CaSi2 particles, then Si-based nanosheets were formed. The morphological, structural and optical properties of the Si-based nanosheets were investigated. It is noted that the thin Si-based nanosheets stacked with a void space formed bundle structures, and the nanosheets were easily exfoliated from the bundles to expose the surfaces corresponding to the Si{111} planes. Meanwhile, the surface of the Si nanosheets might be terminated by O, H, or OH bonds. The Si-based nanosheet bundles were then formed and directly rooted to the Si(111) substrates, and had a remarkably highly symmetrical morphology. This study demonstrated a simple method for preparing Si-based nanosheets, and electro- and photo-chemical applications would possibly be expected, such as in lithium ion batteries.
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