ABSTRACT
Pompia is an ancient name of a small community in Messara valley, which is the main agricultural area, in central Crete. The constructed wetland in Pompia is a free water surface (FWS) system, for treating the wastewater of the local community of 1,200 p.e. That wastewater treatment plant (WWTP) is a pilot plant but it is simple, safe, innovative, and environmentally friendly. The WWTP was funded by the Region of Crete. The Eastern Crete Development Organization was responsible for the design, supervision, management, and initial operation. The project was completed in August 1999, and has been under operation since then. The effluent is considered to be equivalent to tertiary treated municipal wastewater, and it will be used to irrigate olive orchards. The general sense for a visitor is that the FWS system operates like a natural marsh and a habitat of birds and wild animals. In addition, very high removal rates for BOD5, COD, TSS, TKN, TP, TC, and FC have been obtained.
Subject(s)
Conservation of Natural Resources , Ecosystem , Waste Disposal, Fluid/methods , Agriculture , Animals , Animals, Wild , Birds , Cost-Benefit Analysis , Greece , Pilot Projects , Plants , Waste Disposal, Fluid/economics , Water Pollution/prevention & controlABSTRACT
We have previously shown that the mouse heterochromatin protein 1 homologue M31 interacts dynamically with the nuclear envelope. Using quantitative in vitro assays, we now demonstrate that this interaction is potently inhibited by soluble factors present in mitotic and interphase cytosol. As indicated by depletion and order-of-addition experiments, the inhibitory activity co-isolates with a 55-kDa protein, which binds avidly to the nuclear envelope and presumably blocks M31-binding sites. Purification of this protein and microsequencing of tryptic peptides identify it as alpha2/6:beta2-tubulin. Consistent with this observation, bona fide tubulin, isolated from rat brain and maintained in a nonpolymerized state, abolishes binding of M31 to the nuclear envelope and aborts M31-mediated nuclear envelope reassembly in an in vitro system. These observations provide a new example of "moonlighting," a process whereby multimeric proteins switch function when their aggregation state or localization is altered.
Subject(s)
Chromosomal Proteins, Non-Histone/metabolism , Heterochromatin/metabolism , Nuclear Envelope/physiology , Tubulin/metabolism , Amino Acid Sequence , Animals , Brain/metabolism , Chromobox Protein Homolog 5 , Chromosomal Proteins, Non-Histone/chemistry , Endometrial Neoplasms , Female , HeLa Cells , Humans , Kinetics , Mice , Molecular Sequence Data , Molecular Weight , Nuclear Envelope/ultrastructure , Peptide Fragments/chemistry , Protein Binding , Recombinant Proteins/chemistry , Recombinant Proteins/metabolism , Tumor Cells, CulturedABSTRACT
To study the dynamics of mammalian HP1 proteins we have microinjected recombinant forms of mHP1alpha, M31 and M32 into the cytoplasm of living cells. As could be expected from previous studies, the three fusion proteins were efficiently transported into the nucleus and targeted specific chromatin areas. However, before incorporation into these areas the exogenous proteins accumulated in a peripheral zone and associated closely with the nuclear envelope. This transient association did not occur when the cells were treated with deacetylase inhibitors, indicating an acetylation-inhibited interaction. In line with these observations, recombinant HP1 proteins exhibited saturable binding to purified nuclear envelopes and stained the nuclei of detergent-permeabilized cells in a rim-like fashion. Competition experiments with various M31 mutants allowed mapping of the nuclear envelope-binding site within an N-terminal region that includes the chromodomain. A His(6)-tagged peptide representing this region inhibited recruitment of LAP2beta and B-type lamins around the surfaces of condensed chromosomes, suggesting involvement of HP1 proteins in nuclear envelope reassembly.
