The fungal hydrophobin RolA recruits polyesterase and laterally moves on hydrophobic surfaces

Toru Takahashi, Hiroshi Maeda, Sachiyo Yoneda, Shinsaku Ohtaki, Yohei Yamagata, Fumihiko Hasegawa, Katsuya Gomi, Tasuku Nakajima, Keietsu Abe

Research output: Contribution to journalArticle

54 Citations (Scopus)

Abstract

When fungi grow on plant or insect surfaces coated with wax polyesters that protect against pathogens, the fungi generally form aerial hyphae to contact the surfaces. Aerial structures such as hyphae and conidiophores are coated with hydrophobins, which are surface-active proteins involved in adhesion to hydrophobic surfaces. When the industrial fungus Aspergillus oryzae was cultivated in a liquid medium containing the biodegradable polyester polybutylene succinate-coadipate (PBSA), the rolA gene encoding hydrophobin RolA was highly transcribed. High levels of RolA and its localization on the cell surface in the presence of PBSA were confirmed by immunostaining. Under these conditions, A. oryzae simultaneously produced the cutinase CutL1, which hydrolyses PBSA. Pre-incubation of PBSA with RolA stimulated PBSA degradation by CutL1, suggesting that RolA bound to the PBSA surface was required for the stimulation. Immunostaining revealed that PBSA films coated with RolA specifically adsorbed CutL1. Quartz crystal microbalance analyses further demonstrated that RolA attached to a hydrophobic sensor chip specifically adsorbed CutL1. Circular dichroism spectra of soluble-state RolA and bound RolA suggested that RolA underwent a conformational change after its adsorption to hydrophobic surfaces. These results suggest that RolA adsorbed to the hydrophobic surface of PBSA recruits CutL1, resulting in condensation of CutL1 on the PBSA surface and consequent stimulation of PBSA hydrolysis. A fluorescence recovery after photobleaching experiment on PBSA films coated with FITC-labelled RolA suggested that RolA moves laterally on the film. We discuss the novel molecular functions of RolA with regard to plastic degradation.

Original languageEnglish
Pages (from-to)1780-1796
Number of pages17
JournalMolecular Microbiology
Volume57
Issue number6
DOIs
Publication statusPublished - 2005 Sep 1

ASJC Scopus subject areas

  • Microbiology
  • Molecular Biology

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