TY  - JOUR
A1  - Panzer, Sabine
A1  - Zhang, Chong
A1  - Konte, Tilen
A1  - Bräuer, Celine
A1  - Diemar, Anne
A1  - Yogendran, Parathy
A1  - Yu-Strzelczyk, Jing
A1  - Nagel, Georg
A1  - Gao, Shiqiang
A1  - Terpitz, Ulrich
T1  - Modified Rhodopsins From Aureobasidium pullulans Excel With Very High Proton-Transport Rates
JF  - Frontiers in Molecular Biosciences
N2  - Aureobasidium pullulans is a black fungus that can adapt to various stressful conditions like hypersaline, acidic, and alkaline environments. The genome of A. pullulans exhibits three genes coding for putative opsins ApOps1, ApOps2, and ApOps3. We heterologously expressed these genes in mammalian cells and Xenopus oocytes. Localization in the plasma membrane was greatly improved by introducing additional membrane trafficking signals at the N-terminus and the C-terminus. In patch-clamp and two-electrode-voltage clamp experiments, all three proteins showed proton pump activity with maximal activity in green light. Among them, ApOps2 exhibited the most pronounced proton pump activity with current amplitudes occasionally extending 10 pA/pF at 0 mV. Proton pump activity was further supported in the presence of extracellular weak organic acids. Furthermore, we used site-directed mutagenesis to reshape protein functions and thereby implemented light-gated proton channels. We discuss the difference to other well-known proton pumps and the potential of these rhodopsins for optogenetic applications.
KW  - black yeast
KW  - photoreceptor
KW  - microbial rhodopsins
KW  - optogenetics
KW  - proton channel
KW  - membrane trafficking
KW  - fungal rhodopsins
KW  - Aureobasidium
Y1  - 2021
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-249248
SN  - 2296-889X
VL  - 8
ER  - 
TY  - JOUR
A1  - Adam, Alexander
A1  - Deimel, Stephan
A1  - Pardo-Medina, Javier
A1  - García-Martínez, Jorge
A1  - Konte, Tilen
A1  - Limón, M. Carmen
A1  - Avalos, Javier
A1  - Terpitz, Ulrich
T1  - Protein activity of the \(Fusarium\) \(fujikuroi\) rhodopsins CarO and OpsA and their relation to fungus−plant interaction
JF  - International Journal of Molecular Sciences
N2  - Fungi possess diverse photosensory proteins that allow them to perceive different light wavelengths and to adapt to changing light conditions in their environment. The biological and physiological roles of the green light-sensing rhodopsins in fungi are not yet resolved. The rice plant pathogen Fusarium fujikuroi exhibits two different rhodopsins, CarO and OpsA. CarO was previously characterized as a light-driven proton pump. We further analyzed the pumping behavior of CarO by patch-clamp experiments. Our data show that CarO pumping activity is strongly augmented in the presence of the plant hormone indole-3-acetic acid and in sodium acetate, in a dose-dependent manner under slightly acidic conditions. By contrast, under these and other tested conditions, the Neurospora rhodopsin (NR)-like rhodopsin OpsA did not exhibit any pump activity. Basic local alignment search tool (BLAST) searches in the genomes of ascomycetes revealed the occurrence of rhodopsin-encoding genes mainly in phyto-associated or phytopathogenic fungi, suggesting a possible correlation of the presence of rhodopsins with fungal ecology. In accordance, rice plants infected with a CarO-deficient F. fujikuroi strain showed more severe bakanae symptoms than the reference strain, indicating a potential role of the CarO rhodopsin in the regulation of plant infection by this fungus.
KW  - fungal rhodopsins
KW  - CarO
KW  - OpsA
KW  - Fusarium fujikuroi
KW  - Oryza sativa
KW  - rice–plant infection
KW  - green light perception
KW  - indole-3-acetic acid (IAA)
KW  - bakanae
KW  - patch-clamp
Y1  - 2018
U6  - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-285125
SN  - 1422-0067
VL  - 19
IS  - 1
ER  -