An ESCRT–spastin interaction promotes fission of recycling tubules from the endosome

Mechanisms coordinating endosomal degradation and recycling are poorly understood, as are the cellular roles of microtubule (MT) severing. We show that cells lacking the MT-severing protein spastin had increased tubulation of and defective receptor sorting through endosomal tubular recycling compartments. Spastin required the ability to sever MTs and to interact with ESCRT-III (a complex controlling cargo degradation) proteins to regulate endosomal tubulation. Cells lacking IST1 (increased sodium tolerance 1), an endosomal sorting complex required for transport (ESCRT) component to which spastin binds, also had increased endosomal tubulation. Our results suggest that inclusion of IST1 into the ESCRT complex allows recruitment of spastin to promote fission of recycling tubules from the endosome. Thus, we reveal a novel cellular role for MT severing and identify a mechanism by which endosomal recycling can be coordinated with the degradative machinery. Spastin is mutated in the axonopathy hereditary spastic paraplegia. Zebrafish spinal motor axons depleted of spastin or IST1 also had abnormal endosomal tubulation, so we propose this phenotype is important for axonal degeneration.

Biochemical Analyses of Human IST1 and Its Function in Cytokinesis

The newly described yeast endosomal sorting complexes required for transport (ESCRT) protein increased sodium tolerance-1 (Ist1p) binds the late-acting ESCRT proteins Did2p/charged MVB protein (CHMP) 1 and Vps4p and exhibits synthetic vacuolar protein sorting defects when combined with mutations in the Vta1p/LIP5–Vps60p/CHMP5 complex. Here, we report that human IST1 also functions in the ESCRT pathway and is required for efficient abscission during HeLa cell cytokinesis. IST1 binding interactions with VPS4, CHMP1, LIP5, and ESCRT-I were characterized, and the IST1–VPS4 interaction was investigated in detail. Mutational and NMR spectroscopic studies revealed that the IST1 terminus contains two distinct MIT interacting motifs (MIM1 and MIM2) that wrap around and bind in different groves of the MIT helical bundle. IST1, CHMP1, and VPS4 were recruited to the midbodies of dividing cells, and depleting either IST1 or CHMP1 proteins blocked VPS4 recruitment and abscission. In contrast, IST1 depletion did not inhibit human immunodeficiency virus-1 budding. Thus, IST1 and CHMP1 act together to recruit and modulate specific VPS4 activities required during the final stages of cell division.

Impact of high saline wastewaters on anaerobic granular sludge functionalities

Three UASB reactors were operated at different salinity levels in order to assess the effects on the granular sludge properties. High levels of activity inhibition were observed at sodium concentrations over 7 g Na+/L, which resulted in low applicable organic loading rates and VFA accumulation in reactor effluents. However, either sludge adaptation or selection for saline resistant microorganisms occurred, which could be concluded from the observed increase in the 50% activity inhibitory concentrations of sodium during continuous flow experiments. Changes in Na+ susceptibility in time are likely to be expected when treating saline wastewaters. The latter was evidenced by the high sodium tolerance of granular methanogenic sludge coming from a full-scale industrial reactor treating such wastewater. High salinity conditions resulted in a reduced granule strength, predicting process instabilities during long term reactor operation. Batch tests showed that high sodium concentrations seemed to displace the calcium from the granular sludge, a factor known to affect anaerobic granules formation.

Foliar and Root Absorption and Translocation of Bispyribac-sodium in Cool-season Turfgrass

Response of creeping bentgrass, annual bluegrass, and Kentucky bluegrass to foliar, soil, or foliar plus soil applications of bispyribac-sodium was evaluated in greenhouse studies. Soil-alone and foliar plus soil applications of bispyribac-sodium at 148 or 296 g ai/ha resulted in greater injury and shoot dry weight reduction of all species 28 d after treatment (DAT) compared to foliar-alone treatments. Creeping bentgrass was less injured than annual or Kentucky bluegrass regardless of application placement. Further studies evaluated foliar and root absorption and translocation of14C-bispyribac-sodium in creeping bentgrass, annual bluegrass, Kentucky bluegrass, and roughtstalk bluegrass. Foliar absorption into creeping bentgrass was less than that of other species at most harvest timings from 4 to 72 h after treatment. Annual and roughstalk bluegrass translocated greater amounts of foliar-absorbed14C to the crown and shoots compared to creeping bentgrass. Annual and roughstalk bluegrass accumulated approximately 47% more14C per dry weight of plant tissue than creeping bentgrass and 74% more than Kentucky bluegrass after 72 h in nutrient solution containing14C-bispyribac-sodium. Annual and roughstalk bluegrass translocated approximately 80% of root-absorbed14C to shoots, whereas creeping bentgrass and Kentucky bluegrass translocated slightly less (66% of absorbed for both species). These studies suggest that bispyribac-sodium is readily absorbed by roots and translocated to shoots which may contribute to its total activity within a plant. In addition, creeping bentgrass displayed lower amounts of foliar and root absorption and subsequent translocation than annual and roughstalk bluegrass which may contribute to greater bispyribac-sodium tolerance displayed by creeping bentgrass.