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Prochlorococcus phage ferredoxin: structural characterization and electron transfer to cyanobacterial sulfite reductases

Marine cyanobacteria are infected by phages whose genomes encode ferredoxin (Fd) electron carriers. These Fds are thought to redirect the energy harvested from light to phage-encoded oxidoreductases that enhance viral fitness, but it is unclear how the biophysical properties and partner specificities of phage Fds relate to those of photosynthetic organisms. Here, results of a bioinformatics analysis using a sequence similarity network revealed that phage Fds are most closely related to cyanobacterial Fds that transfer electrons from photosystems to oxidoreductases involved in nutrient assimilation. Structural analysis of myovirus P-SSM2 Fd (pssm2-Fd), which infects the cyanobacterium Prochlorococcus marinus, revealed high levels of similarity to cyanobacterial Fds (root mean square deviations of ≤0.5 Å). Additionally, pssm2-Fd exhibited a low midpoint reduction potential (–336 mV versus a standard hydrogen electrode), similar to other photosynthetic Fds, although it had lower thermostability (Tm = 28 °C) than did many other Fds. When expressed in an Escherichia coli strain deficient in sulfite assimilation, pssm2-Fd complemented bacterial growth when coexpressed with a P. marinus sulfite reductase, revealing that pssm2-Fd can transfer electrons to a host protein involved in nutrient assimilation. The high levels of structural similarity with cyanobacterial Fds and reactivity with a host sulfite reductase suggest that phage Fds evolved to transfer electrons to cyanobacterially encoded oxidoreductases.

cyanobacteria

Microbial Adaptation to Spaceflight Is Correlated With Bacteriophage-Encoded Functions

Evidence from the International Space Station suggests microbial populations are rapidly adapting to the spacecraft environment; however, the mechanism of this adaptation is not understood. Bacteriophages are prolific mediators of bacterial adaptation on Earth. Here we survey 245 genomes sequenced from bacterial strains isolated on the International Space Station for dormant (lysogenic) bacteriophages. Our analysis indicates phage-associated genes are significantly different between spaceflight strains and their terrestrial counterparts. Additionally, we identify 283 complete prophages, those that could initiate bacterial lysis and infect additional hosts, of which 21% are novel. These prophage regions encode functions that correlate with increased persistence in extreme environments, such as spaceflight, to include antimicrobial resistance and virulence, DNA damage repair, and dormancy. Our results correlate microbial adaptation in spaceflight to bacteriophage-encoded functions that may impact human health in spaceflight.

Space Biology

Investigation of Prophage Regions of Bacterial Strains Isolated from the International Space Station (ISS)

Space flight agencies are planning missions back to the Moon and to Mars. When sending humans into space, it is impossible to separate them from microorganisms, either in their associated microbiome or in the spacecraft environment, which are modified through the movement of genetic material. Bacteriophages, small viruses that invade and replicate within bacterial cells, play a central role in the genetic composition and evolution of microorganisms. Lysogenic bacteriophages can insert themselves into the DNA of their bacterial hosts, forming prophage regions, which can also transfer genes from previous hosts. Thus, we aimed to identify and classify all prophages from twelve bacterial species cultured from the International Space Station (ISS) from 2017 to 2018. We determined representative bacterial strains for each species, whose genomes were analyzed to identify prophage regions. Complete prophages were identified through database searches and the number of prophage regions were compared to terrestrial analogs. Additionally, prophage region and genome sizes were compared for each species, identifying the percentage of bacteriophage DNA in each genome. We determined that the prophage-susceptible bacterial species isolated from the ISS had a higher number of prophage regions when compared to terrestrial analogs, as well as having a larger percentage of their genomes made up of prophage material. Of the eighteen complete prophages identified, 72.2% were of family Siphoviridae and 27.8% were of family Myoviridae. Only one of the prophages had a BLAST similarity over 80%, suggesting that the remainder of prophages are novel species. These results imply that there is a higher rate of prophage transduction and lysogeny during spaceflight, and that the prophages present are novel.

Phage

Clustered DNA damages induced in isolated DNA and in human cells by low doses of ionizing radiation

Clustered DNA damages-two or more closely spaced damages (strand breaks, abasic sites, or oxidized bases) on opposing strands-are suspects as critical lesions producing lethal and mutagenic effects of ionizing radiation. However, as a result of the lack of methods for measuring damage clusters induced by ionizing radiation in genomic DNA, neither the frequencies of their production by physiological doses of radiation, nor their repairability, nor their biological effects are known. On the basis of methods that we developed for quantitating damages in large DNAs, we have devised and validated a way of measuring ionizing radiation-induced clustered lesions in genomic DNA, including DNA from human cells. DNA is treated with an endonuclease that induces a single-strand cleavage at an oxidized base or abasic site. If there are two closely spaced damages on opposing strands, such cleavage will reduce the size of the DNA on a nondenaturing gel. We show that ionizing radiation does induce clustered DNA damages containing abasic sites, oxidized purines, or oxidized pyrimidines. Further, the frequency of each of these cluster classes is comparable to that of frank double-strand breaks; among all complex damages induced by ionizing radiation, double-strand breaks are only about 20%, with other clustered damage constituting some 80%. We also show that even low doses (0.1-1 Gy) of high linear energy transfer ionizing radiation induce clustered damages in human cells.

NASA Discipline Radiation Health