The formation mechanism of apical sterol rich membrane domains (SRDs) and visualization of SRDs by Photoactivated Localization Microscopy in Aspergillus nidulans

Ref ID: 18474

Author:

Norio Takeshita, Yuji Ishitsuka, Ulrich Nienhaus, Reinhard Fischer

Author address:

Karlsruhe Institute of Technology (KIT), 1Institute for Applied Biosciences, Institute for Applied Physics, Karlsruhe,
Germany

Full conference title:

11 th European Conference on Fungal Genetics

Abstract:

Apical sterol8208;rich plasma membrane domains (SRDs), which can be viewed using the sterol8208;binding fluorescent dye
filipin, are gaining attention for their important roles in polarized growth of filamentous fungi. The microdomain
scaffolding protein flotillin was thought to be a good candidate involved in the formation of SRDs. We analyzed the
function of the flotillin orthologue FloA by gene deletion and protein localization in the maintenance of SRDs and
polarity. SRDs are known to be necessary for the localization of some components of the growth machinery. To
investigate deeply the relation of lipid membrane domains and protein localization, the distribution of
microdomains in SRDs are analyzed by super8208;resolution microscope technique, Photoactivated Localization
Microscopy (PALM). Raft membranes and non8208;raft membranes were visualized by each marker protein tagged
with photoconvertible fluorescent protein mEosFP for PALM. The size of SRDs is around a few 61549;m, whereas the
size of lipid rafts ranges in general between 108208;200 nm. In recent years, super8208;resolution microscope techniques
have been improving and breaking the diffraction limit of conventional light microscopy whose resolution limit is
250 nm. In this method, a lateral image resolution as high as 20 nm will be a powerful tool to investigate
membrane microdomains.

Abstract Number: PR1.79

Conference Year: 2012

Link to conference website: http://www.ecfg.info/Abstract_submission.php

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