Mechanism of sphingolipid homeostasis revealed by structural analysis of \u3ci\u3eArabidopsis\u3c/i\u3e SPT-ORM1 complex

The serine palmitoyltransferase (SPT) complex catalyzes the first and rate-limiting step in sphingolipid biosynthesis in all eukaryotes. ORM/ORMDL proteins are negative regulators of SPT that respond to cellular sphingolipid levels. However, the molecular basis underlying ORM/ORMDL-dependent homeostatic regulation of SPT is not well understood.We determined the cryo–electron microscopy structure of Arabidopsis SPT-ORM1 complex, composed of LCB1, LCB2a, SPTssa, and ORM1, in an inhibited state. A ceramide molecule is sandwiched between ORM1 and LCB2a in the cytosolic membrane leaflet. Ceramide binding is critical for the ORM1-dependent SPT repression, and dihydroceramides and phytoceramides differentially affect this repression. A hybrid β sheet, formed by the amino termini of ORM1 and LCB2a and induced by ceramide binding, stabilizes the amino terminus of ORM1 in an inhibitory conformation. Our findings provide mechanistic insights into sphingolipid homeostatic regulation via the binding of ceramide to the SPT-ORM/ORMDL complex that may have implications for plant-specific processes such as the hypersensitive response for microbial pathogen resistance

KULL: LLNL's ASCI Inertial Confinement Fusion Simulation Code

KULL is a three dimensional, time dependent radiation hydrodynamics simulation code under development at Lawrence Livermore National Laboratory. A part of the U.S. Department of Energy's Accelerated Strategic Computing Initiative (ASCI), KULL's purpose is to simulate the physical processes in Inertial Confinement Fusion (ICF) targets. The National Ignition Facility, where ICF experiments will be conducted, and ASCI are part of the experimental and computational components of DOE's Stockpile Stewardship Program. This paper provides an overview of ASCI and describes KULL, its hydrodynamic simulation capability and its three methods of simulating radiative transfer. Particular emphasis is given to the parallelization techniques essential to obtain the performance required of the Stockpile Stewardship Program and to exploit the massively parallel processor machines that ASCI is procuring

Collaborative regulation of yeast SPT-Orm2 complex by phosphorylation and ceramide

Summary: The homeostatic regulation of serine palmitoyltransferase (SPT) activity in yeast involves N-terminal phosphorylation of Orm proteins, while higher eukaryotes lack these phosphorylation sites. Although recent studies have indicated a conserved ceramide-mediated feedback inhibition of the SPT-ORM/ORMDL complex in higher eukaryotes, its conservation and relationship with phosphorylation regulation in yeast remain unclear. Here, we determine the structure of the yeast SPT-Orm2 complex in a dephosphomimetic state and identify an evolutionarily conserved ceramide-sensing site. Ceramide stabilizes the dephosphomimetic Orm2 in an inhibitory conformation, facilitated by an intramolecular β-sheet between the N- and C-terminal segments of Orm2. Moreover, we find that a phosphomimetic mutant of Orm2, positioned adjacent to its intramolecular β-sheet, destabilizes the inhibitory conformation of Orm2. Taken together, our findings suggest that both Orm dephosphorylation and ceramide binding are crucial for suppressing SPT activity in yeast. This highlights a distinctive regulatory mechanism in yeast involving the collaborative actions of phosphorylation and ceramide