Single-molecule speckle analysis of actin filament turnover in lamellipodia

Lamellipodia are thin, veil-like extensions at the edge of cells that contain a dynamic array of actin filaments. We describe an approach for analyzing spatial regulation of actin polymerization and depolymerization in vivo in which we tracked single molecules of actin fused to the green fluorescent protein. Polymerization and the lifetime of actin filaments in lamellipodia were measured with high spatial precision. Basal polymerization and depolymerization occurred throughout lamellipodia with largely constant kinetics, and polymerization was promoted within one micron of the lamellipodium tip. Most of the actin filaments in the lamellipodium were generated by polymerization away from the tip.     

Protostar formation in the early universe

The nature of the first generation of stars in the universe remains largely unknown. Observations imply the existence of massive primordial stars early in the history of the universe, and the standard theory for the growth of cosmic structure predicts that structures grow hierarchically through gravitational instability. We have developed an ab initio computer simulation of the formation of primordial stars that follows the relevant atomic and molecular processes in a primordial gas in an expanding universe. The results show that primeval density fluctuations left over from the Big Bang can drive the formation of a tiny protostar with a mass 1% that of the Sun. The protostar is a seed for the subsequent formation of a massive primordial star.     

Rotational movement of the formin mDia1 along the double helical strand of an actin filament

Formin homology proteins (formins) elongate actin filaments (F-actin) by continuously associating with filament tips, potentially harnessing actin-generated pushing forces. During this processive elongation, formins are predicted to rotate along the axis of the double helical F-actin structure (referred to here as helical rotation), although this has not yet been definitively shown. We demonstrated helical rotation of the formin mDia1 by single-molecule fluorescence polarization (FL(P)). FL(P) of labeled F-actin, both elongating and depolymerizing from immobilized mDia1, oscillated with a periodicity corresponding to that of the F-actin long-pitch helix, and this was not altered by actin-bound nucleotides or the actin-binding protein profilin. Thus, helical rotation is an intrinsic property of formins. To harness pushing forces from growing F-actin, formins must be anchored flexibly to cell structures.     

Actin polymerization-driven molecular movement of mDia1 in living cells

mDia1, a Rho effector, belongs to the Formin family of proteins, which shares the conserved tandem FH1-FH2 unit structure. Formins including mDia1 accelerate actin nucleation while interacting with actin filament fast-growing ends. Here our single-molecule imaging revealed fast directional movement of mDia1 FH1-FH2 for tens of microns in living cells. The movement of mDia1 FH1-FH2 was blocked by actin-perturbing drugs, and the speed of mDia1 FH1-FH2 movement appeared to correlate with actin elongation rates. In vitro, mDia1 FH1-FH2 associated persistently with the growing actin barbed end. mDia1 probably moves processively along the growing end of actin filaments in cells, and Formins may be a molecular motility machinery that is independent from motor proteins.     

Gigantic photoresponse in 1/4-filled-band organic salt (EDO-TTF)2PF6

We report that the organic salt (EDO-TTF)2PF6 with 3/4-filled-band (1/4-filled in terms of holes), which forms an organic metal with strong electron and lattice correlation, shows a highly sensitive response to photoexcitation. An ultrafast, photoinduced phase transition from the insulator phase to the metal phase can be induced with very weak excitation intensity at near room temperature. This response makes the material attractive for applications in switching devices with room-temperature operation. The observed photo-induced spectroscopic change shows that this photoinduced phase transition process is caused by the cooperative melting of charge ordering assisted by coherent phonon generation.     

Uracil DNA glycosylase activity is dispensable for immunoglobulin class switch

Activation-induced cytidine deaminase (AID) is required for the DNA cleavage step in immunoglobulin class switch recombination (CSR). AID is proposed to deaminate cytosine to generate uracil (U) in either mRNA or DNA. In the second instance, DNA cleavage depends on uracil DNA glycosylase (UNG) for removal of U. Using phosphorylated histone gamma-H2AX focus formation as a marker of DNA cleavage, we found that the UNG inhibitor Ugi did not inhibit DNA cleavage in immunoglobulin heavy chain (IgH) locus during CSR, even though Ugi blocked UNG binding to DNA and strongly inhibited CSR. Strikingly, UNG mutants that had lost the capability of removing U rescued CSR in UNG-/- B cells. These results indicate that UNG is involved in the repair step of CSR yet by an unknown mechanism. The dispensability of U removal in the DNA cleavage step of CSR requires a reconsideration of the model of DNA deamination by AID.     

Lodging resistance locus prl5 improves physical strength of the lower plant part under different conditions of fertilization in rice (Oryza sativa L.)

High fertilizer application rates increase grain yield and biomass in rice, but reduces lodging resistance. Lodging resistance locus prl5 enhances pushing resistance through delaying leaf senescence and increasing carbohydrate reaccumulation in stems. This study demonstrates the effects of prl5 under different levels of Nitrogen·Phosphorus·Potassium basal fertilizer application using a chromosome segment substitution line based on the genetic background of Koshihikari, which contains prl5. High fertilizer application rates enhanced pushing resistance of the lower part, but did not consistently affect the pushing resistance of the whole plant. Independent of fertilizer application rates, prl5 enhanced pushing resistances of both the lower part and the whole plants. Plant height, biomass and grain yield were increased by high fertilizer application rates, but stem diameter and the dry weight of basal culms did not show consistent increases as determined over a period of 2 years. In leaves just after heading, chlorophyll (Chl) contents, rather than Rubisco contents, were correlated with fertilizer application rates. At the fully ripe stage, fertilizer application did not affect Chl and Rubisco contents in leaves, while prl5 increased the contents in the second leaf. High fertilizer application rates did not affect the concentration of non-structural carbohydrates (NSCs) in basal culms, but prl5 increased them at the fully ripe stage. These results suggest that fertilizer application reduces lodging resistance by highly elevating the center of gravity, but that prl5 can counteract this effect by enhancing the physical strength of the lower part through delaying leaf senescence and increasing NSC reaccumulation in culms without any effects of different fertilizer application rates.