Functional analysis of the AAA ATPase AipA localizing at the endocytic sites in the filamentous fungus Aspergillus oryzae.

Ref ID: 15835

Author:

Yujiro Higuchi,

Author address:

Manabu Arioka, Katsuhiko Kitamoto. Department of Biotechnology, The University of Tokyo, Japan.

Full conference title:

26th Fungal Genetics Conference

Date: 15 March 2014

Abstract:

We explored novel components involved in endocytosis by the yeast two-hybrid (YTH) screening using AoAbp1 (Aspergillus oryzae actin binding
protein) as bait. A gene named aipA (AoAbp1 interacting protein) which encodes a putative AAA (ATPases associated with diverse cellular activities)
ATPase was obtained. Further YTH analyses showed that 346-370 aa region of AipA interacted with the two SH3 domains of AoAbp1. AipA interacted
with AoAbp1 in vitro, and in A. oryzae EGFP-AipA co-localized with AoAbp1-mDsRed at the tip region, suggesting that AipA functions in endocytosis.
Although aipA disruptants did not display any phenotypic alteration in several culture conditions, aipA-overexpressing strains showed defective growth
and the aberrant hyphal morphology. Moreover, we generated strains which have mutations of either aipAK542A or aipAE596Q. These mutations were
introduced in the ATPase domain of AipA and would cause defect in the ATPase activity. In contrast to the strain overexpressing WT aipA, the growth
of mutated strains was normal, suggesting that ATPase activity is important for the function of AipA. Furthermore, the aipA-overexpressing strain
displayed a delay in FM4- 64 transport to Spitzenkorper, whereas the mutated aipA-overexpressing strains did not, suggesting that AipA negatively
regulates apical endocytic recycling.

Abstract Number: NULL

Conference Year: 2011

Link to conference website: NULL

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