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A Genetic Screen to Isolate Toxoplasma gondii Host-cell Egress Mutants
Published on: February 8, 2012
The invasion pore induced by Toxoplasma gondii
Yuto Kegawa1, Frances Male2, Irene Jiménez-Munguía1
1Section on Integrative Biophysics; Division of Basic and Translational Biophysics, Eunice Kennedy Shriver National Institute of Child Health and Human Development (NICHD), National Institutes of Health (NIH), Bethesda, MD, USA.
None:
The parasite Toxoplasma gondii invades its host cell only after secreting proteins such as invasion-requisite RON2 that inserts into the host cell membrane to establish the moving junction. Electrophysiological recordings at sub-200 µs resolution show a transient increase in host cell membrane conductance following parasite exposure. Transients always precede invasion, but parasites depleted of RON2 generate transients without invading. Thus RON2 is not essential for transient generation. Time-series analysis developed here and applied to the 910,000 data point transient dataset reveal multiple quantal conductance changes in the parasite-induced transient, consistent with rapid insertion, then slower removal, blocking, or inactivation of potential pore components. Quantal steps for wild-type RH strain parasites have a principal mode with Gaussian mean of 0.26 nS, similar in step size to the pore forming protein EXP2, part of the PTEX translocon of malaria parasites. Without RON2 the quantal mean (0.19 nS) is significantly different. Because we observe no parasite invasion without poration, the term "invasion pore" is proposed to describe this transient breach in host cell membrane barrier integrity during invasion.
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