Coordinated regulation and complex formation of yellow variegated1 and yellow variegated2, chloroplastic FtsH metalloproteases involved in the repair cycle of photosystem II in Arabidopsis thylakoid membranes

Arabidopsis yellow variegated1 (VAR1) and VAR2 are separate loci that encode similar chloroplast FtsH proteases. To date, FtsH is the best-characterized protease in thylakoid membranes involved in the turnover of photosynthetic protein complexes. It comprises a protein family that is encoded by 12 different nuclear genes in Arabidopsis. We show here that nine FtsH proteins are located in the chloroplasts. Mutations in either VAR1 or VAR2 cause typical leaf variegation and sensitivity to photoinhibition. By contrast, none of these phenotypes was observed in T-DNA insertion mutants in other ftsH genes (ftsh1, ftsh6, and ftsh8) closely related to VAR1 and VAR2. This finding suggests that VAR1 and VAR2 play a predominant role in the photosystem II repair cycle in thylakoid membranes. By generating VAR1- and VAR2-specific antibodies, we found that loss of either VAR1 or VAR2 results in the decreased accumulation of the other. Thus, the genetic nonredundancy between VAR1 and VAR2 could be attributed to their coordinated regulation at the protein level. These observations led us to examine whether VAR1 and VAR2 form a complex. Sucrose density gradient and gel filtration analyses revealed a complex of approximately 400 to 450 kD, probably representing a hexamer. Furthermore, VAR1 and VAR2 were shown to coprecipitate by immunoprecipitation using VAR1- and VAR2-specific antibodies. The majority of VAR1 appears to exist as heterocomplexes with VAR2, whereas VAR2 may be present as homocomplexes as well. Based on these results, we conclude that VAR1 and VAR2 are the major components of an FtsH complex involved in the repair of photodamaged proteins in thylakoid membranes.     

Biological productivity of Lake Towada,a north temperate,oligotrophic, kokanee fishery lake

During the Lake Towada survey from April through October 1998, we measured primary production at shore and offshore stations, and calculated crustacean zooplankton production from samples collected at the offshore station. We then analyzed these data and compared them with commercial fishery data in order to discuss the energy flow in this lake where kokanee (Oncorhynchus nerka) fisheries are one of the main businesses. At all stations, primary production was relatively high: 150–300mgCm^–2day^–1 in April–mid-June and lower at ca. 100mgCm^–2day^–1 thereafter. The variation in primary productivity could largely be explained by multiple regression models that included phytoplankton biomass and ambient nutrient conditions as independent variables. Among zooplankton, rotifers had their peak abundance in May, before the crustacean zooplankton (Daphnia longispina, Bosmina longirostris, and Acanthodiaptomus pacificus) population was well established. D. longispina dominated the crustacean zooplankton community in terms of biomass and production; their production during the study period made up 80% of crustacean community production (19.6gdry-wtm^–2), which was 40% of primary production during the survey. In July, when the abundance of D. longispina was particularly high, their daily production slightly exceeded daily primary production, which resulted in ca. 30% and 75% reduction in the amount of particulate organic carbon and chlorophyll a, respectively, during this period. The community ingestion rate of crustacean zooplankton, calculated from their daily production (D. longispina accounted for 90%) and the assumption that their gross production efficiency (K₁) was 60%, could roughly explain this reduction of particulate matter, corroborating previous studies that the grazing of D. longispina can significantly improve the water transparency of this lake. The catch of kokanee and pond smelt (Hypomesus transpacificus nipponensis, another important fish in the lake) during the survey corresponded to 1.1% of crustacean community production, and corresponded to 0.45% of the primary production, which is one of the highest recorded values. Bearing in mind that D. longispina was the major food item of planktivorous fish such as kokanee and pond smelt, the present study suggests that the energy transfer from phytoplankton to zooplankton to fish is outstandingly efficient, compared with other aquatic ecosystems, when D. longispina dominates in the lake.     

Synthetic studies on guanofosfocin: glycosylation of 8-oxo-purine nucleosides via Mitsunobu reaction

In model studies directed to the total synthesis of guanofosfocins, a unique glycosidic bond formation between the 8-oxo-purine nucleosides and mannopyranose derivatives under Mistunobu conditions is described.     

Cholesteryl glucoside-induced protection against gastric ulcer

The cytoprotective effect of heat shock proteins (HSPs) promises new therapeutic modalities for medical treatment. We examined the anti-ulcer effect of cholesteryl glucoside (1-O-cholesteryl-beta-D-glucopyranoside, CG) on cold-restraint stress-induced gastric ulcer in rats, in terms of its correlative ability to activate heat shock factor (HSF) and to induce HSP70. Rapid induction of CG occurred in animal tissues, especially in stomach, after exposure to stress, indicating that this glycolipid might act as an anti-stress, lipid mediator involved in the very early stages of stress-induced signal transduction. Orally administered CG apparently showed anti-ulcer activity in rats via HSF activation and HSP70 induction. When compared with geranylgeranylacetone (GGA), the well known as an effective, synthetic anti-ulcer agent, CG proved to have the same level of strength on ulcer inhibition. GGA caused CG and HSP70 induction in gastric mucosa, indicating that GGA induced HSP70 via CG production. CG thus might be useful for medical treatment of stress-induced diseases, and as an anti-stress supplement for daily diet.     

Crystal structure of AlgQ2, a macromolecule (alginate)-binding protein of Sphingomonas sp. A1 at 2.0A resolution

Sphingomonas sp. A1 possesses a high molecular mass (average 25,700 Da) alginate uptake system mediated by a novel pit-dependent ABC transporter. The X-ray crystallographic structure of AlgQ2 (57,200 Da), an alginate-binding protein in the system, was determined by the multiple isomorphous replacement method and refined at 2.0 A resolution with a final R-factor of 18.3% for 15 to 2.0 A resolution data. The refined structure of AlgQ2 was comprised of 492 amino acid residues, 172 water molecules, and one calcium ion. AlgQ2 was composed of two globular domains with a deep cleft between them, which is expected to be the alginate-binding site. The overall structure is basically similar to that of maltose/maltodextrin-binding protein, except for the presence of an N2-subdomain. The entire calcium ion-binding site is similar to the site in the EF-hand motif, but comprises a ten residue loop. This calcium ion-binding site is about 40 A away from the alginate-binding site.