Another Really, Really Big Virus

Viruses with genomes larger than 300 kb and up to 1.2 Mb, which encode hundreds of proteins, are being discovered and characterized with increasing frequency. Most, but not all, of these large viruses (often referred to as giruses) infect protists that live in aqueous environments. Bioinformatic analyses of metagenomes of aqueous samples indicate that large DNA viruses are quite common in nature and await discovery. One issue that is perhaps not appreciated by the virology community is that large viruses, even those classified in the same family, can differ significantly in morphology, lifestyle, and gene complement. This brief commentary, which will mention some of these unique properties, was stimulated by the characterization of the newest member of this club, virus CroV (Fischer, M.G.; Allen, M.J.; Wilson, W.H.; Suttle, C.A. Giant virus with a remarkable complement of genes infects marine zooplankton. Proc. Natl. Acad. Sci. USA 2010, 107, 19508–19513 [1]). CroV has a 730 kb genome (with ∼544 protein-encoding genes) and infects the marine microzooplankton Cafeteria roenbergensis producing a lytic infection

PROMOTERS FROM CHLORELLAVIRUS GENES PROVIDING EXPRESSION OF GENES IN PROKARYOTIC AND EUKARYOTIC HOSTS

The invention is directed to novel promoters or mutants thereof from Chlorella virus DNA methyltransferase genes. A Chlorella virus gene promoter is operably linked to a first and/or second DNA sequence encoding a gene that is different from the Chlorella virus gene to form an expression cassette. An expression cassette can be introduced into prokaryotic and/or eukaryotic cells and can provide for a high level of expression of the gene encoded by the first and/or second DNA sequence. The invention also provides a method for screening other Chlorella virus genes for promoters that can function to express a heterologous gene in prokaryotic and/or eukaryotic hosts

Synthesis and Localization of a Development-Specific Protein in Sclerotia of \u3ci\u3eSclerotinia sclerotiorum\u3c/i\u3e

A development-specific protein (SSP) makes up about 35 to 40% of the total protein in sclerotia of the fungus Sclerotinia sclerotiorum. The protein consists of three charge isomers, with one isomer making up 80 to 90% of the total. In vitro translation of poly(A)+ RNA isolated from cells in early stages of sclerotia formation revealed that 44% of the amino acids incorporated was into SSP. In vivo- and in vitro-synthesized forms of SSP migrated at identical rates on both isoelectric focusing and denaturing polyacrylamide gels, indicating that SSP was not synthesized as a larger precursor. This was significant because SSP accumulated in membrane-bound, organellelike structures which resemble protein bodies found in seeds of many higher plants

Biochemical Changes During the Growth of Fungi: I. Nitrogen Compounds and Carbohydrate Changes in \u3ci\u3ePenicillium atrovenetum\u3c/i\u3e

Changes in the biochemical constituents of cells were studied during the growth and development of Penicillium atrovenetum. Growth of the fungus, as measured by the dry weight, could be divided into four phases: lag, log, stationary, and death. The percentages of total nitrogen, cold trichloroacetic acid-soluble nitrogen, ribonucleic acid (RNA), and protein increased to a maximum during the lag phase, and subsequently decreased as the fungus aged. The percentage of deoxyribonucleic acid (DNA) was always slightly higher in the spores than in the mycelium. The DNA in the mycelium decreased in the lag phase, and then increased slightly to a plateau for the duration of the log phase, followed by a decrease to a constant percentage during the stationary and death phases. Carbohydrates were present in higher concentration in the mycelium than in the spores. The percentage of carbohydrates in the mycelium increased continually until it reached a maximum late in the log phase, and then decreased as the fungus entered the death phase. The results reported for this fungus are, in general, in agreement with those reported for other microorganisms. Namely, the percentages of enzyme-forming compounds, such as amino acids, nucleotides, RNA, and protein, were highest in the lag phase, whereas storage compounds such as carbohydrates increased to a maximum near the end of the log phase. The definition of log phase in fungi depends on the criteria that are used. If, instead of using the linear increase in dry weight to delimit this growth period, one uses the end of net protein, RNA, and DNA synthesis, a more realistic concept of growth emerges

PROMOTERS FROM CHLORELLA VIRUS GENES PROVIDING FOR EXPRESSION OF GENES IN PROKARYOTIC AND EUKARYOTIC HOSTS

The invention is directed to novel promoters or mutants thereof from Chlorella virus DNA methyltansferase genes. A Chlorella Virus gene promoter is operably linked to a first and/or Second DNA sequence encoding a gene that is different from the Chlorella virus to form an expression cassette. An expression cassette can be introduced into prokaryotic and/or eukaryotic cells and can provide for a high level of expression of the gene encoded by the first and/or Second DNA sequence. The invention also provides a method for Screening other Chlorella virus genes for promoters that can function to express a heterologous gene in prokaryotic and/or eukaryotic hosts

Biography of James L. Van Etten

Green algae, in surface layers of almost every lake or stream, are some of the most common aquatic creatures. However, unbeknownst to researchers until recently, viruses that infect algae are almost as widespread. Entire ecosystems of algal hosts and their corresponding viruses lay hidden until the 1980s, when James L. Van Etten, a professor of plant pathology at the University of Nebraska (Lincoln), and his colleague Russ Meints discovered and began to characterize the first member of what is now a rapidly expanding family of algal viruses. Van Etten and his colleagues have continued to study these intriguing viruses, focusing on those that infect Chlorella and other similar green algae. The chlorella viruses have many unusual properties, ranging from their large genome sizes to unique modifications in their DNA

Biography of James L. Van Etten

Green algae, in surface layers of almost every lake or stream, are some of the most common aquatic creatures. However, unbeknownst to researchers until recently, viruses that infect algae are almost as widespread. Entire ecosystems of algal hosts and their corresponding viruses lay hidden until the 1980s, when James L. Van Etten, a professor of plant pathology at the University of Nebraska (Lincoln), and his colleague Russ Meints discovered and began to characterize the first member of what is now a rapidly expanding family of algal viruses. Van Etten and his colleagues have continued to study these intriguing viruses, focusing on those that infect Chlorella and other similar green algae. The chlorella viruses have many unusual properties, ranging from their large genome sizes to unique modifications in their DNA

Synthesis and Localization of a Development-Specific Protein in Sclerotia of \u3ci\u3eSclerotinia sclerotiorum\u3c/i\u3e

A development-specific protein (SSP) makes up about 35 to 40% of the total protein in sclerotia of the fungus Sclerotinia sclerotiorum. The protein consists of three charge isomers, with one isomer making up 80 to 90% of the total. In vitro translation of poly(A)+ RNA isolated from cells in early stages of sclerotia formation revealed that 44% of the amino acids incorporated was into SSP. In vivo- and in vitro-synthesized forms of SSP migrated at identical rates on both isoelectric focusing and denaturing polyacrylamide gels, indicating that SSP was not synthesized as a larger precursor. This was significant because SSP accumulated in membrane-bound, organellelike structures which resemble protein bodies found in seeds of many higher plants

Giant Chloroviruses: Five Easy Questions

Chloroviruses are large, icosahedral, dsDNA-containing viruses that replicate in certain unicellular, chlorella-like green algae [1,2]. They exist in freshwater throughout the world with titers as high as thousands of plaque-forming units (PFU) per ml of indigenous water although titers are typically 1–100 PFU/ml. Titers fluctuate during the year with the highest titers typically occurring in the spring and late fall. Known chlorovirus hosts, which are normally symbionts and are often referred to as zoochlorellae, are associated with either the protozoan Paramecium bursaria (Fig 1A), the coelenterate Hydra viridis, or the heliozoan Acanthocystis turfacea. Zoochlorellae are resistant to viruses in their symbiotic state. Fortunately, some zoochlorellae grow independently of their partners in the laboratory, permitting plaque assay of the viruses (Fig 1B) and synchronous infection of their hosts, which allows one to study the virus life cycle in detail

The \u3ci\u3ePhycodnaviridae\u3c/i\u3e: The Story of How Tiny Giants Rule the World

The family Phycodnaviridae encompasses a diverse and rapidly expanding collection of large icosahedral, dsDNA viruses that infect algae. These lytic and lysogenic viruses have genomes ranging from 160 to 560 kb. The family consists of six genera based initially on host range and supported by sequence comparisons. The family is monophyletic with branches for each genus, but the phycodnaviruses have evolutionary roots that connect them with several other families of large DNA viruses, referred to as the nucleocytoplasmic large DNA viruses (NCLDV).The phycodnaviruses have diverse genome structures, some with large regions of noncoding sequence and others with regions of ssDNA. The genomes of members in three genera in the Phycodnaviridae have been sequenced. The genome analyses have revealed more than 1000 unique genes, with only 14 homologous genes in common among the three genera of phycodnaviruses sequenced to date. Thus, their gene diversity far exceeds the number of so-called core genes. Not much is known about the replication of these viruses, but the consequences of these infections on phytoplankton have global affects, including influencing geochemical cycling and weather patterns