The function of a contractile vacuole, or CV, is to control the balance of intracellular water and act as osmoregulatory organelles for amoebae and protozoa. A cell must have a certain amount of solute and solvent to function properly. Here, water becomes a solvent and must be proportional to an additional material or solute (source). In order to identify possible regulators and effectors of Disgorgin, we purified a complex containing disgorgine and analyzed mass spectrometry (additional figure S4A). None of the proteins discussed above have been identified in the Disgorgin complex. We found the FpaA and FpaB orthologous SKP1, the components of the ubiquitination complex SCF, ubiquitin and the vacuolar subunit H+-ATPase A VatA. Co-immunoprecipitation experiments confirmed the in vivo interaction of disgorgine, FpaA and FpaB in a manner dependent on the F-box domain (additional figure S4B). These results will be discussed in the discussion and additional data. A contractile vacuole (CV) is an organelle or subcellular structure involved in osmoregulation and waste disposal. Previously, a CV was known as a pulsed or pulsed vacuole. CVs should not be confused with vacuoles that store food or water. A CV is mainly found in protists and unicellular algae. In freshwater environments, the concentration of solutes inside the cell is higher than outside the cell.

Under these conditions, water flows from the environment into the cell by osmosis. Thus, the CV acts as a protective mechanism against cell expansion (and possibly explosion) due to too much water; It expels excess water from the cell by contracting. However, not all species that have a CV are freshwater organisms; some marine and soil microorganisms also have a CV. VC is predominant in species that do not have a cell wall, but there are exceptions. During the evolutionary process, CV was mainly eliminated in multicellular organisms; However, there are still several multicellular fungi in the single-celled stage and in various types of cells in sponges, including amoebocytes, pinacocytes and choanocytes. Table 2.6 — Phenotypes of contractile vacuole mutants (CV) Disgorgin locates in the CV membrane. (A) Localization of endogenous disgorgine and V-ATPase (VatM). The co-location of Disgorgin and V-ATPase is indicated by arrows. (B) Localization of GFP disgorging with dajumine RFP in Ax2 cells. The co-location of Disgorgin and Dajumin is indicated by arrows.

(C) Location of dajumine RFP (left panel) or GFP disgorgine (right panel) in Ax2 cells under hypotonic stress. The images were taken for both colors at the same time. Full arrows indicate a discharge event and open arrows indicate a full loading and unloading cycle. (D) Cell morphology of disgorginous cells or Ax2 cells expressing truncated disgorgine. The arrows indicate enlarged CVs. (E) Localization of truncated disgorgine in disgorgine cells or cells Ax2 and FM4-64 in cells GFP-C382−717/disgorgin−. To better illustrate the location, an additional image of GFP-C382−717/disgorgin− is shown. The arrows indicate the vacuoles where GFP-C382−717 and FM4-64 are located. Ladder rods, 5 μm. To put it simply, a contractile vacuole expels fluid during contraction in some protozoa. As a contractile vacuole, the subcellular structure works in parallel with osmoregulation, which occurs in protists and single-celled algae.

It has also been called pulsating or pulsed vacuole. The contractile vacuole complex / osmoregulatory system of the ciliates consists of a contractile vacuole and emitting radial channels that pass through a firmly attached tubular “spongioma”. It has two preformed cyclic fusion/membrane cleavage sites. These are the “porus” for excretion of exocytotic fluid and connection of radial channels with the vacuole (Allen and Naitoh, 2002). Electrophysiology has determined its periodic dis-/reconnection from/to the vacuole by capacity measurements (Tominaga et al., 1998a,b). Exactly these sites, as well as the docking site of the vacuole to the cell membrane, are labeled with antibodies against NSF, provided that the dissociation of NSF is inhibited in carefully permeabilized cells by adding the NSF inhibitor, N-ethyl maleimide and non-hydrolyzable ATP γ-S (Kissmehl et al., 2002). Note that NSF is normally released by membrane anocking/fusion sites at each cycle (Section 3.2). Although the biogenesis of the contractile vacuol system has not yet been clarified, a preliminary interpretation of the reported facts is as follows. Based on section 3.3, it can be assumed that the assembly of the components of the vacuol contractile complex would begin in the emergency room, followed by delivery by Golgi-derived vesicles.

Within the spongioma, lateral segregation can be imagined, perhaps made possible by the tendency of H+ ATPase molecules to form higher-order dimers and linear clusters (Strauss et al., 2008). This, in turn, could lead to segregation in the decorated spongioma and this process could lead to its tubularization (Allen et al., 1989). Although Disgorgin is a Rab8A-GAP, overexpression of Rab8A in Disgorgin cells suppresses the stationary accumulation of large vacuoles (that is, large vacuoles do not accumulate; Data not shown), possibly by providing a sufficient level of Rab8A GDP to the CV system, which could compete with Rab8A-GTP for CV membrane sites. However, Rab8A does not suppress the abnormal discharge of CV into disgorginous cells (data not shown), suggesting that the transition from the GTP-related form to the GDP-related form of Rab8A is important for fulfilling its function in the fusion of the CV plasma membrane. Protozoa containing parasitic elements live in an isotonic environment for their internal environment or cytoplasm. .