title: The fungal hydrophobin RolA recruits polyesterase and laterally moves on hydrophobic surfaces.
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.
authors.affiliation: Laboratory of Enzymology, Division of Life Science, Graduate School of Agricultural Science, Tohoku University, Sendai 985-8555, Japan.
authors.forename: Toru
authors.initials: T
authors.lastname: Takahashi
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Maeda
authors.forename: Sachiyo
authors.initials: S
authors.lastname: Yoneda
authors.forename: Shinsaku
authors.initials: S
authors.lastname: Ohtaki
authors.forename: Yohei
authors.initials: Y
authors.lastname: Yamagata
authors.forename: Fumihiko
authors.initials: F
authors.lastname: Hasegawa
authors.forename: Katsuya
authors.initials: K
authors.lastname: Gomi
authors.forename: Tasuku
authors.initials: T
authors.lastname: Nakajima
authors.forename: Keietsu
authors.initials: K
authors.lastname: Abe
mesh_terms: D000272:Adipates / Q000378:metabolism*; D000327:Adsorption; D001236:Aspergillus oryzae / Q000235:genetics / Q000254:growth & development / Q000378:metabolism* / Q000502:physiology; D001673:Biodegradation, Environmental; D002265:Carboxylic Ester Hydrolases / Q000235:genetics / Q000378:metabolism; D002942:Circular Dichroism; D003470:Culture Media; D004950:Esterases / Q000378:metabolism*; D005656:Fungal Proteins / Q000235:genetics / Q000378:metabolism* / Q000502:physiology; D015966:Gene Expression Regulation, Fungal; D057927:Hydrophobic and Hydrophilic Interactions; D008856:Microscopy, Fluorescence; D011091:Polyesters / Q000378:metabolism; D011487:Protein Conformation; D013386:Succinates / Q000378:metabolism*
chemical_list: D000272:Adipates; D003470:Culture Media; D005656:Fungal Proteins; D011091:Polyesters; D013386:Succinates; C437084:poly(tetramethylene succinate-co-tetramethylene adipate); D004950:Esterases; D002265:Carboxylic Ester Hydrolases; C019952:cutinase