Translocation of an 89-kDa periplasmic protein is associated with Holospora infection

Fate of the 63-kDa periplasmic protein of the infectious form of the endonuclear symbiotic bacteriumHolospora obtusaduring the infection process

“…For 2D SDS‐PAGE, 2 × 10 8 IF cells were loaded, whereas 6 × 10 6 IF cells and 5 × 10 7 RF cells were loaded in each lane for 1D SDS‐PAGE. The gels were then transferred to membranes and incubated overnight with the prospective mAbs as described previously (Fujishima et al , 1990; Iwatani et al , 2005).…”

“…An equal volume of culture medium was added 3 days after mixing. Aliquots (5 mL) of the cell suspension were collected from the mixture at various times after mixing, fixed in 4% paraformaldehyde in phosphate‐buffered saline (PBS) (Iwatani et al , 2005) for 10 min, and then subjected to IIFM without air‐drying.…”

“…To assay culture supernatants of hybridoma cells, aliquots of IF cells were air‐dried on cover glasses (4.5 mm × 24 mm), fixed with 4% paraformaldehyde for 10 min, permeabilized with 20 mM NaOH for 45 s (Iwatani et al , 2005), and washed twice with PBS. The cells were consecutively incubated with mAbs and Alexa Fluor 488 (AF488) goat anti‐mouse IgG (Molecular Probes Inc.) diluted 2000‐fold with PBS, for 3 h at 25 °C or overnight at 4 °C, washed with PBS, stained with 0.25 μg mL −1 4′,6‐diamidino‐2‐phenylindole dihydrochloride (DAPI), and observed under differential‐interference‐contrast (DIC) and fluorescence microscopes (BX60, Olympus, Tokyo, Japan; TSC SL, Leica).…”

“…Dohra et al (1994) developed monoclonal antibodies (mAbs) against 39‐ and 15‐kDa periplasmic proteins of the IF cells and illustrated the qualitative and quantitative changes of these proteins during the infection process (Fujishima et al , 1997). In addition, we recently demonstrated the important role of another 89‐kDa periplasmic protein localized in the invasion tip during bacterial escape from the host digestive vacuole, migration through the host cytoplasm towards the target nucleus, and invasion into the macronucleus (Iwatani et al , 2005). The present study was designed to find another periplasmic protein that plays a significant role in the infection process.…”

Response of the bacterial symbiont Holospora caryophila to different growth conditions of its host

“…presence versus absence of the connecting piece and different symbiont maintenance strategies during host's sexual processes. It should be also remembered that the molecular and functional bases of the relationship between H. caryophila and its host are substantially unexplored, contrary to the classical holosporas (Abamo et al 2008;Iwatani et al 2005;Sabaneyeva et al, 2009;Wiemann and Görtz, 1991), for which also genome projects have been undertaken Dohra et al 2013). Analogue investigations on H. caryophila would clearly benefit from an optimisation of growth conditions, which can be obtained as an application of the findings from our study and its further developments.…”