Crystal Structure of Reovirus Attachment Protein σ1 in Complex with Sialylated Oligosaccharides

Many viruses attach to target cells by binding to cell-surface glycans. To gain a better understanding of strategies used by viruses to engage carbohydrate receptors, we determined the crystal structures of reovirus attachment protein σ1 in complex with α-2,3-sialyllactose, α-2,6-sialyllactose, and α-2,8-di-siallylactose. All three oligosaccharides terminate in sialic acid, which serves as a receptor for the reovirus serotype studied here. The overall structure of σ1 resembles an elongated, filamentous trimer. It contains a globular head featuring a compact β-barrel, and a fibrous extension formed by seven repeating units of a triple β-spiral that is interrupted near its midpoint by a short α -helical coiled coil. The carbohydrate-binding site is located between β-spiral repeats two and three, distal from the head. In all three complexes, the terminal sialic acid forms almost all of the contacts with σ1 in an identical manner, while the remaining components of the oligosaccharides make little or no contacts. We used this structural information to guide mutagenesis studies to identify residues in σ1 that functionally engage sialic acid by assessing hemagglutination capacity and growth in murine erythroleukemia cells, which require sialic acid binding for productive infection. Our studies using σ1 mutant viruses reveal that residues 198, 202, 203, 204, and 205 are required for functional binding to sialic acid by reovirus. These findings provide insight into mechanisms of reovirus attachment to cell-surface glycans and contribute to an understanding of carbohydrate binding by viruses. They also establish a filamentous, trimeric carbohydrate-binding module that could potentially be used to endow other trimeric proteins with carbohydrate-binding properties

Alfalfa-grass biomass, soil organic carbon, and total nitrogen under different management approaches in an irrigated agroecosystem

Background and aims Management approach may influence forage production as well as soil organic carbon (SOC) and soil total nitrogen (STN) accrued beneath perennial grass-legume components of irrigated crop rotations. This study aimed to evaluate effects of conventional, certified organic, and reduced-tillage management approaches on above- and belowground biomass production and C and N content in alfalfa-grass mixture, and their relationships with SOC and STN. Methods An alfalfa-grass mixture was established in 2009 on four replications under a sprinkler irrigation system. Soil characteristics were analyzed at planting time in 2009. Aboveground biomass production, coarse and fine roots, SOC, STN, aboveground biomass C and N, and coarse- and fine-root C and N were quantified in samples collected during 2009–2011. Results Conventional management produced more aboveground biomass than reduced-tillage and organic, but production under organic matched conventional and exceeded reduced-tillage in the last two harvests of the study. Root production was constant under the three approaches, but resulted in more SOC accrued under reduced-tillage than under the other two approaches. Conclusions Biomass production was favored by conventional seedbed preparation and soil fertility management while SOC accrual was favored by minimum soil disturbance. In addition, aboveground biomass was influenced by seasonal air temperature, precipitation, and nutrient mineralization from the previous season, soabove-/belowground allocation changed seasonally