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From the Drosophila virilis late puff region 31C, we microcloned two neighbouring genes, Kil-1 and Kil-2, that encode putative Kunitz serine protease inhibitor like proteins. The Kil-1 gene is expressed exclusively in prepupal salivary glands. Using a size mutant of the KIL-1 protein and MALDI-TOF analysis, we demonstrate that during pupation this protein is released from the prepupal salivary glands into the pupation fluid covering the surface of the pupa. 3-D- structure predictions are consistent with the known crystal structure of the human Kunitz type protease inhibitor 2KNT. This is the first experimental proof for the extra-corporal presence of a distinct Drosophila prepupal salivary gland protein. Possible functions of KIL-1 in the context of the control of proteolytic activities in the pupation fluid are discussed. (C) 2004 Elsevier Ltd. All rights reserved
A putative phosphatase, LSF1 (for LIKE SEX4; previously PTPKIS2), is closely related in sequence and structure to STARCH-EXCESS4 (SEX4), an enzyme necessary for the removal of phosphate groups from starch polymers during starch degradation in Arabidopsis (Arabidopsis thaliana) leaves at night. We show that LSF1 is also required for starch degradation: lsf1 mutants, like sex4 mutants, have substantially more starch in their leaves than wild-type plants throughout the diurnal cycle. LSF1 is chloroplastic and is located on the surface of starch granules. lsf1 and sex4 mutants show similar, extensive changes relative to wild-type plants in the expression of sugar-sensitive genes. However, although LSF1 and SEX4 are probably both involved in the early stages of starch degradation, we show that LSF1 neither catalyzes the same reaction as SEX4 nor mediates a sequential step in the pathway. Evidence includes the contents and metabolism of phosphorylated glucans in the single mutants. The sex4 mutant accumulates soluble phospho- oligosaccharides undetectable in wild-type plants and is deficient in a starch granule-dephosphorylating activity present in wild-type plants. The lsf1 mutant displays neither of these phenotypes. The phenotype of the lsf1/sex4 double mutant also differs from that of both single mutants in several respects. We discuss the possible role of the LSF1 protein in starch degradation.
The hypersaline crater lake and its catchment on seabird island Isabel (Pacific, off Mexico) was studied to explore the influence of strong seasonal variations in rainfall/evaporation and guano contribution on its limnology. The hypersaline lake water (HSW, 78 %) is up to 2.2-times enriched in inert ions relative to mean seawater. Rainfall during summer dilutes the HSW to form a less saline rainwater body (RWB) above a chemolimnion between 2 and 4 m water depth. The RWB is inhabited first by diatoms and ostracods followed later on by cyanobacteria and ciliates. Evaporation of > 1.5 m depth of lake water over the dry season increases the salinity of the RWB until the water column becomes isohaline at HSW concentrations in the late dry season. Differences in the stable isotope composition of water and primary producers in RWB and HSW reflect this development. Introduction of seabird guano and the decrease of salinity fuel a high primary production in the RWB with higher delta(CDIC)-C-13 and delta(13)Corg of particulate organic matter than in the HSW. The high N supply leads to high delta N-15 NH4 values (+ 39 % in the HSW) as the consequence of ammonia volatilization that is strongest during guano maturation and with evaporative salinity increase from the HSW. Precipitation of carbonate (calcite and aragonite) from the RWB and the HSW is hindered by the high concentration of guano-derived P. This inhibition may be overcome with evaporative supersaturation during particularly dry conditions. Carbonate may also precipitate during particularly wet conditions from the dilute RWB, where the P-concentration is reduced during an active phytoplankton production that raises the pH. Differences in the stable isotope signatures of carbon and oxygen in HSW and RWB (+ 5 % delta(CDIC)-C-13 and -3 % d18OH2O) suggest the processes of carbonate precipitation can be distinguished based on the isotope signature of the carbonates deposited. Changes in the lake system are indicated when lower temperatures and higher rainfall in the 2006 wet season introduced more and less mature guano to the lake. The lower pH was accompanied by lower ammonia volatilization and carbonate precipitation as indicated by an increased concentration of NH4, Ca, Sr and DIC, while delta H-2, delta(NNH4)-N-15, and salinity were lower. According to our results, the observed sediment laminations should reflect the introduction of catchment material (including guano) with runoff, the RWB plankton production, and the carbonate precipitation in relation to its origin and seasonality.