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1 May 2002 The Vacuolar-ATPase of Paramecium multimicronucleatum:Gene Structure of the B Subunit and the Dynamics of theV-ATPase-rich Osmoregulatory Membranes
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Abstract

Previous studies have shown that the vacuolar-ATPase (V-ATPase) of the contractile vacuole complexes (CVCs) in Paramecium multimicronucleatum is necessary for fluid segregation and osmoregulation. In the current study, immunofluorescence showed that the development of a new CVC begins with the formation of a new pore around which the collecting canals form. The decorated membranes are then deposited around the newly formed collecting canals. Quick-freeze deep-etch techniques reveal that six 10-nm-wide V-ATPase V1 sectors, tightly packed into a 20 × 30-nm rectangle, form two rows of these compacted sectors that helically wrap around the cytosolic side of decorated membrane tubules. During new CVC formation, packing of decorated tubules around mature CVCs was temporarily disrupted so that some of these decorated tubules became transformed into decorated vesicles. Freeze-fracturing of these decorated vesicles revealed a highly pitted E-face and a particulate P-face. The V-ATPase was purified for the first time in any ciliated protozoan and shown to contain, as in other cells, the V1 subunits A to E, and four 14–20 kDa polypeptides. The B subunit was cloned and found to be encoded by one gene containing four short introns. This subunit has 510 amino acid residues with a predicted molecular weight of 56.8 kDa, a value similar to B subunits of other organisms. Except for the N- and C-termini, it has a 75% sequence identity with other B subunits, suggesting that the B subunits in Paramecium, like other species, have been conserved and that the entire surface of this subunit may be important in interacting with other subunits.

AGNES K. FOK, KIYOSHI YAMAUCHI, AKINORI ISHIHARA, MARILYNN S. AIHARA, MASAKI ISHIDA, and RICHARD D. ALLEN "The Vacuolar-ATPase of Paramecium multimicronucleatum:Gene Structure of the B Subunit and the Dynamics of theV-ATPase-rich Osmoregulatory Membranes," The Journal of Eukaryotic Microbiology 49(3), 185-196, (1 May 2002). https://doi.org/10.1111/j.1550-7408.2002.tb00521.x
Received: 1 December 2001; Accepted: 23 February 2002; Published: 1 May 2002
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