TY  - JOUR
AU  - Stevanović, Katarina S
AU  - Čepkenović, Bogdana
AU  - Križak, Strahinja
AU  - Pajić, Tanja
AU  - Todorović, Nataša
AU  - Živić, Miroslav Ž
PY  - 2023
UR  - http://radar.ibiss.bg.ac.rs/handle/123456789/6088
AB  - Ion channels are vital components of filamentous fungi signaling in communication with their environment. We exploited the ability of the apical region of growing sporangiophores of Phycomyces blakesleeanus to form membrane-enveloped cytoplasmic droplets (CDs), to examine ion currents in the filamentous fungi native plasma membrane. In hypoosmotic conditions, the dominant current in the CDs is ORIC, an osmotically activated, anionic, outwardly rectified, fast inactivating instantaneous current that we have previously characterized. Here, we examined the effect of ATP on ORIC. We show that CDs contain active mitochondria, and that respiration inhibition by azide accelerates ORIC inactivation. ATP, added intracellularly, reduced ORIC run-down and shifted the voltage dependence of inactivation toward depolarized potentials, in a manner that did not require hydrolysis. Notably, ATP led to slowing down of ORIC inactivation, as evidenced by an increased time constant of inactivation, τin, and slower decline of τin during prolonged recordings. Flavonoids (genistein and quercetin) had the effect on ORIC opposite to ATP, acting as current inhibitors, possibly by disrupting the stabilizing effect of ATP on ORIC. The integration of osmotic sensing with ATP dependence of the anionic current, typical of vertebrate cells, is described here for the first time in filamentous fungi.
PB  - Springer Nature
T2  - Scientific Reports
T1  - ATP modulation of osmotically activated anionic current in the membrane of Phycomyces blakesleeanus sporangiophore
IS  - 1
VL  - 13
DO  - 10.1038/s41598-023-39021-9
SP  - 11897
ER  - 

@article{
author = "Stevanović, Katarina S and Čepkenović, Bogdana and Križak, Strahinja and Pajić, Tanja and Todorović, Nataša and Živić, Miroslav Ž",
year = "2023",
abstract = "Ion channels are vital components of filamentous fungi signaling in communication with their environment. We exploited the ability of the apical region of growing sporangiophores of Phycomyces blakesleeanus to form membrane-enveloped cytoplasmic droplets (CDs), to examine ion currents in the filamentous fungi native plasma membrane. In hypoosmotic conditions, the dominant current in the CDs is ORIC, an osmotically activated, anionic, outwardly rectified, fast inactivating instantaneous current that we have previously characterized. Here, we examined the effect of ATP on ORIC. We show that CDs contain active mitochondria, and that respiration inhibition by azide accelerates ORIC inactivation. ATP, added intracellularly, reduced ORIC run-down and shifted the voltage dependence of inactivation toward depolarized potentials, in a manner that did not require hydrolysis. Notably, ATP led to slowing down of ORIC inactivation, as evidenced by an increased time constant of inactivation, τin, and slower decline of τin during prolonged recordings. Flavonoids (genistein and quercetin) had the effect on ORIC opposite to ATP, acting as current inhibitors, possibly by disrupting the stabilizing effect of ATP on ORIC. The integration of osmotic sensing with ATP dependence of the anionic current, typical of vertebrate cells, is described here for the first time in filamentous fungi.",
publisher = "Springer Nature",
journal = "Scientific Reports",
title = "ATP modulation of osmotically activated anionic current in the membrane of Phycomyces blakesleeanus sporangiophore",
number = "1",
volume = "13",
doi = "10.1038/s41598-023-39021-9",
pages = "11897"
}

Stevanović, K. S., Čepkenović, B., Križak, S., Pajić, T., Todorović, N.,& Živić, M. Ž.. (2023). ATP modulation of osmotically activated anionic current in the membrane of Phycomyces blakesleeanus sporangiophore. in Scientific Reports
Springer Nature., 13(1), 11897.
https://doi.org/10.1038/s41598-023-39021-9

Stevanović KS, Čepkenović B, Križak S, Pajić T, Todorović N, Živić MŽ. ATP modulation of osmotically activated anionic current in the membrane of Phycomyces blakesleeanus sporangiophore. in Scientific Reports. 2023;13(1):11897.
doi:10.1038/s41598-023-39021-9 .

Stevanović, Katarina S, Čepkenović, Bogdana, Križak, Strahinja, Pajić, Tanja, Todorović, Nataša, Živić, Miroslav Ž, "ATP modulation of osmotically activated anionic current in the membrane of Phycomyces blakesleeanus sporangiophore" in Scientific Reports, 13, no. 1 (2023):11897,
https://doi.org/10.1038/s41598-023-39021-9 . .

TY  - JOUR
AU  - Stevanović, Katarina
AU  - Čepkenović, Bogdana
AU  - Križak, Strahinja
AU  - Živić, Miroslav
AU  - Todorović, Nataša
PY  - 2023
UR  - http://radar.ibiss.bg.ac.rs/handle/123456789/5978
AB  - Studies of ion currents in filamentous fungi are a prerequisite for forming a complete
understanding of their physiology. Cytoplasmic droplets (CDs), obtained from sporangiophores of
Phycomyces blakesleeanus, are a model system that enables the characterization of ion currents in
the native membrane, including the currents mediated by the channels not yet molecularly identified.
Osmotically activated anionic current with outward rectification (ORIC) is a dominant current in
the membrane of cytoplasmic droplets under the conditions of hypoosmotic stimulation. We have
previously reported remarkable functional similarities of ORIC with the vertebrate volume regulated
anionic current (VRAC), such as dose-dependent activation by osmotic difference, ion selectivity
sequence, and time and voltage dependent profile of the current. Using the patch clamp method on
the CD membrane, we further resolve VRAC-like ORIC characteristics in this paper. We examine
the inhibition by extracellular ATP and carbenoxolone, the permeation of glutamate in presence of
chloride, selectivity for nitrates, and activation by GTP, and we show its single channel behavior
in excised membrane. We propose that ORIC is a functional counterpart of vertebrate VRAC in
filamentous fungi, possibly with a similar essential role in anion efflux during cell volume regulation.
PB  - Basel: MDPI
T2  - Journal of Fungi
T1  - Osmotically activated anion current of Phycomyces blakesleeanus-filamentous fungi counterpart to vertebrate volume regulated anion current
IS  - 6
VL  - 9
DO  - 10.3390/jof9060637
SP  - 637
ER  - 

@article{
author = "Stevanović, Katarina and Čepkenović, Bogdana and Križak, Strahinja and Živić, Miroslav and Todorović, Nataša",
year = "2023",
abstract = "Studies of ion currents in filamentous fungi are a prerequisite for forming a complete
understanding of their physiology. Cytoplasmic droplets (CDs), obtained from sporangiophores of
Phycomyces blakesleeanus, are a model system that enables the characterization of ion currents in
the native membrane, including the currents mediated by the channels not yet molecularly identified.
Osmotically activated anionic current with outward rectification (ORIC) is a dominant current in
the membrane of cytoplasmic droplets under the conditions of hypoosmotic stimulation. We have
previously reported remarkable functional similarities of ORIC with the vertebrate volume regulated
anionic current (VRAC), such as dose-dependent activation by osmotic difference, ion selectivity
sequence, and time and voltage dependent profile of the current. Using the patch clamp method on
the CD membrane, we further resolve VRAC-like ORIC characteristics in this paper. We examine
the inhibition by extracellular ATP and carbenoxolone, the permeation of glutamate in presence of
chloride, selectivity for nitrates, and activation by GTP, and we show its single channel behavior
in excised membrane. We propose that ORIC is a functional counterpart of vertebrate VRAC in
filamentous fungi, possibly with a similar essential role in anion efflux during cell volume regulation.",
publisher = "Basel: MDPI",
journal = "Journal of Fungi",
title = "Osmotically activated anion current of Phycomyces blakesleeanus-filamentous fungi counterpart to vertebrate volume regulated anion current",
number = "6",
volume = "9",
doi = "10.3390/jof9060637",
pages = "637"
}

Stevanović, K., Čepkenović, B., Križak, S., Živić, M.,& Todorović, N.. (2023). Osmotically activated anion current of Phycomyces blakesleeanus-filamentous fungi counterpart to vertebrate volume regulated anion current. in Journal of Fungi
Basel: MDPI., 9(6), 637.
https://doi.org/10.3390/jof9060637

Stevanović K, Čepkenović B, Križak S, Živić M, Todorović N. Osmotically activated anion current of Phycomyces blakesleeanus-filamentous fungi counterpart to vertebrate volume regulated anion current. in Journal of Fungi. 2023;9(6):637.
doi:10.3390/jof9060637 .

Stevanović, Katarina, Čepkenović, Bogdana, Križak, Strahinja, Živić, Miroslav, Todorović, Nataša, "Osmotically activated anion current of Phycomyces blakesleeanus-filamentous fungi counterpart to vertebrate volume regulated anion current" in Journal of Fungi, 9, no. 6 (2023):637,
https://doi.org/10.3390/jof9060637 . .

TY  - CONF
AU  - Stevanović, Katarina
AU  - Križak, Strahinja
AU  - Todorović, Nataša
AU  - Živić, Miroslav
PY  - 2018
UR  - http://radar.ibiss.bg.ac.rs/handle/123456789/6375
AB  - Jonski kanali filamentoznih gljiva su još uvek u velikoj meri neistraženi, uglavnom usled prisustva brzo regenerišućeg ćelijskog zida. Naš model sistem citoplazmatskih kapi dobijenih iz sporangiofora gljive Phycomyces blakesleeanus, odnosno ćelijske membrane koja još uvek nije pokrivena ćelijskim zidom, nam omogućava da registrujemo struje metodom nametnute voltaže na deliću membrane. U hipoosmotskim uslovima, pored izlazno ispravljene anjonske struje (ORIC), u familiji struja sa cele membrane su uočljive i ulazno ispravljene anjonske struje koje su propustljive za glukonat. Ovde će biti opisane po prvi put. U hipoosmotskoj sredini (kada je prisutan i ORIC), sa hlorom simetrično zamenjenim glukonatom, glukonat propustljiva struja se javlja u 50% eksperimenata (n=10). U izoosmotskim uslovima, kada ORIC nije aktivan, ulazna struja je 2.5 puta veća kada je simetrični glukonat provodni jon nego kada je prisutan hlor (n=11, prisutna u 72% zapisa). Fit voltažne zavisnosti struje Bolcmanovom funkcijom pokazuje jasno ulazno ispravljanje, sa vrednošću nalektrisanja vratnica -0.55±0.10 (p<0.001). Upadljiva osobina glukonat provodljive struje je spora deaktivacija i odložena aktivacija (na potencijalu -150 mV struja se aktivira posle 137±36 ms (n=8)). Snimanje aktivnosti pojedinačnih kanala u asimetričnim anjonskim uslovima (glukonat/hlor) pokazalo je da u membrani citoplazmatske kapi postoje najmanje tri različita jonska kanala (u 3 od 30 eksperimenata) koji su propustljiviji za glukonat nego za hlor sa sledećim karakteristikama: provodljivost (u pS) 83, 60 i 75; potencijal reverzije (mV) -13,13, 9,42, -5,28; propustljivost za glukonat/propustljivost za hlor 1.80, 1.53, 1.27.
PB  - Belgrade: Serbian Biological Society
C3  - Drugi kongres biologa Srbije; 2018 Sep 25-30; Kladovo, Srbija
T1  - Ulazno ispravljena anjonska struja u membrani citoplazmatskih kapi iz gljive Phycomyces blakesleeanus
SP  - 34
UR  - https://hdl.handle.net/21.15107/rcub_ibiss_6375
ER  - 

@conference{
author = "Stevanović, Katarina and Križak, Strahinja and Todorović, Nataša and Živić, Miroslav",
year = "2018",
abstract = "Jonski kanali filamentoznih gljiva su još uvek u velikoj meri neistraženi, uglavnom usled prisustva brzo regenerišućeg ćelijskog zida. Naš model sistem citoplazmatskih kapi dobijenih iz sporangiofora gljive Phycomyces blakesleeanus, odnosno ćelijske membrane koja još uvek nije pokrivena ćelijskim zidom, nam omogućava da registrujemo struje metodom nametnute voltaže na deliću membrane. U hipoosmotskim uslovima, pored izlazno ispravljene anjonske struje (ORIC), u familiji struja sa cele membrane su uočljive i ulazno ispravljene anjonske struje koje su propustljive za glukonat. Ovde će biti opisane po prvi put. U hipoosmotskoj sredini (kada je prisutan i ORIC), sa hlorom simetrično zamenjenim glukonatom, glukonat propustljiva struja se javlja u 50% eksperimenata (n=10). U izoosmotskim uslovima, kada ORIC nije aktivan, ulazna struja je 2.5 puta veća kada je simetrični glukonat provodni jon nego kada je prisutan hlor (n=11, prisutna u 72% zapisa). Fit voltažne zavisnosti struje Bolcmanovom funkcijom pokazuje jasno ulazno ispravljanje, sa vrednošću nalektrisanja vratnica -0.55±0.10 (p<0.001). Upadljiva osobina glukonat provodljive struje je spora deaktivacija i odložena aktivacija (na potencijalu -150 mV struja se aktivira posle 137±36 ms (n=8)). Snimanje aktivnosti pojedinačnih kanala u asimetričnim anjonskim uslovima (glukonat/hlor) pokazalo je da u membrani citoplazmatske kapi postoje najmanje tri različita jonska kanala (u 3 od 30 eksperimenata) koji su propustljiviji za glukonat nego za hlor sa sledećim karakteristikama: provodljivost (u pS) 83, 60 i 75; potencijal reverzije (mV) -13,13, 9,42, -5,28; propustljivost za glukonat/propustljivost za hlor 1.80, 1.53, 1.27.",
publisher = "Belgrade: Serbian Biological Society",
journal = "Drugi kongres biologa Srbije; 2018 Sep 25-30; Kladovo, Srbija",
title = "Ulazno ispravljena anjonska struja u membrani citoplazmatskih kapi iz gljive Phycomyces blakesleeanus",
pages = "34",
url = "https://hdl.handle.net/21.15107/rcub_ibiss_6375"
}

Stevanović, K., Križak, S., Todorović, N.,& Živić, M.. (2018). Ulazno ispravljena anjonska struja u membrani citoplazmatskih kapi iz gljive Phycomyces blakesleeanus. in Drugi kongres biologa Srbije; 2018 Sep 25-30; Kladovo, Srbija
Belgrade: Serbian Biological Society., 34.
https://hdl.handle.net/21.15107/rcub_ibiss_6375

Stevanović K, Križak S, Todorović N, Živić M. Ulazno ispravljena anjonska struja u membrani citoplazmatskih kapi iz gljive Phycomyces blakesleeanus. in Drugi kongres biologa Srbije; 2018 Sep 25-30; Kladovo, Srbija. 2018;:34.
https://hdl.handle.net/21.15107/rcub_ibiss_6375 .

Stevanović, Katarina, Križak, Strahinja, Todorović, Nataša, Živić, Miroslav, "Ulazno ispravljena anjonska struja u membrani citoplazmatskih kapi iz gljive Phycomyces blakesleeanus" in Drugi kongres biologa Srbije; 2018 Sep 25-30; Kladovo, Srbija (2018):34,
https://hdl.handle.net/21.15107/rcub_ibiss_6375 .

TY  - JOUR
AU  - Stanić, Marin
AU  - Križak, Strahinja
AU  - Jovanović, Mirna
AU  - Pajić, Tanja
AU  - Ćirić, Ana
AU  - Žižić, Milan
AU  - Zakrzewska, Joanna
AU  - Cvetić Antić, Tijana
AU  - Todorović, Nataša
AU  - Živić, Miroslav
PY  - 2017
UR  - http://www.microbiologyresearch.org/content/journal/micro/10.1099/mic.0.000429
UR  - https://radar.ibiss.bg.ac.rs/handle/123456789/2730
AB  - Increasing resistance of fungal strains to known fungicides has prompted identification of new candidates for fungicides among substances previously used for other purposes. We have tested the effects of known anion channel inhibitors anthracene-9-carboxylic acid (A9C) and niflumic acid (NFA) on growth, energy metabolism and anionic current of mycelium of fungus Phycomyces blakesleeanus. Both inhibitors significantly decreased growth and respiration of mycelium, but complete inhibition was only achieved by 100 and 500 μM NFA for growth and respiration, respectively. A9C had no effect on respiration of human NCI-H460 cell line and very little effect on cucumber root sprout clippings, which nominates this inhibitor for further investigation as a potential new fungicide. Effects of A9C and NFA on respiration of isolated mitochondria of P. blakesleeanus were significantly smaller, which indicates that their inhibitory effect on respiration of mycelium is indirect. NMR spectroscopy showed that both A9C and NFA decrease the levels of ATP and polyphosphates in the mycelium of P. blakesleeanus, but only A9C caused intracellular acidification. Outwardly rectifying, fast inactivating instantaneous anionic current (ORIC) was also reduced to 33±5 and 21±3% of its pre-treatment size by A9C and NFA, respectively, but only in the absence of ATP. It can be assumed from our results that the regulation of ORIC is tightly linked to cellular energy metabolism in P. blakesleeanus, and the decrease in ATP and polyphosphate levels could be a direct cause of growth inhibition.
T2  - Microbiology
T1  - Growth inhibition of fungus Phycomyces blakesleeanus by anion channel inhibitors anthracene-9-carboxylic and niflumic acid attained through decrease in cellular respiration and energy metabolites
IS  - 3
VL  - 163
DO  - 10.1099/mic.0.000429
SP  - 364
EP  - 372
ER  - 

@article{
author = "Stanić, Marin and Križak, Strahinja and Jovanović, Mirna and Pajić, Tanja and Ćirić, Ana and Žižić, Milan and Zakrzewska, Joanna and Cvetić Antić, Tijana and Todorović, Nataša and Živić, Miroslav",
year = "2017",
abstract = "Increasing resistance of fungal strains to known fungicides has prompted identification of new candidates for fungicides among substances previously used for other purposes. We have tested the effects of known anion channel inhibitors anthracene-9-carboxylic acid (A9C) and niflumic acid (NFA) on growth, energy metabolism and anionic current of mycelium of fungus Phycomyces blakesleeanus. Both inhibitors significantly decreased growth and respiration of mycelium, but complete inhibition was only achieved by 100 and 500 μM NFA for growth and respiration, respectively. A9C had no effect on respiration of human NCI-H460 cell line and very little effect on cucumber root sprout clippings, which nominates this inhibitor for further investigation as a potential new fungicide. Effects of A9C and NFA on respiration of isolated mitochondria of P. blakesleeanus were significantly smaller, which indicates that their inhibitory effect on respiration of mycelium is indirect. NMR spectroscopy showed that both A9C and NFA decrease the levels of ATP and polyphosphates in the mycelium of P. blakesleeanus, but only A9C caused intracellular acidification. Outwardly rectifying, fast inactivating instantaneous anionic current (ORIC) was also reduced to 33±5 and 21±3% of its pre-treatment size by A9C and NFA, respectively, but only in the absence of ATP. It can be assumed from our results that the regulation of ORIC is tightly linked to cellular energy metabolism in P. blakesleeanus, and the decrease in ATP and polyphosphate levels could be a direct cause of growth inhibition.",
journal = "Microbiology",
title = "Growth inhibition of fungus Phycomyces blakesleeanus by anion channel inhibitors anthracene-9-carboxylic and niflumic acid attained through decrease in cellular respiration and energy metabolites",
number = "3",
volume = "163",
doi = "10.1099/mic.0.000429",
pages = "364-372"
}

Stanić, M., Križak, S., Jovanović, M., Pajić, T., Ćirić, A., Žižić, M., Zakrzewska, J., Cvetić Antić, T., Todorović, N.,& Živić, M.. (2017). Growth inhibition of fungus Phycomyces blakesleeanus by anion channel inhibitors anthracene-9-carboxylic and niflumic acid attained through decrease in cellular respiration and energy metabolites. in Microbiology, 163(3), 364-372.
https://doi.org/10.1099/mic.0.000429

Stanić M, Križak S, Jovanović M, Pajić T, Ćirić A, Žižić M, Zakrzewska J, Cvetić Antić T, Todorović N, Živić M. Growth inhibition of fungus Phycomyces blakesleeanus by anion channel inhibitors anthracene-9-carboxylic and niflumic acid attained through decrease in cellular respiration and energy metabolites. in Microbiology. 2017;163(3):364-372.
doi:10.1099/mic.0.000429 .

Stanić, Marin, Križak, Strahinja, Jovanović, Mirna, Pajić, Tanja, Ćirić, Ana, Žižić, Milan, Zakrzewska, Joanna, Cvetić Antić, Tijana, Todorović, Nataša, Živić, Miroslav, "Growth inhibition of fungus Phycomyces blakesleeanus by anion channel inhibitors anthracene-9-carboxylic and niflumic acid attained through decrease in cellular respiration and energy metabolites" in Microbiology, 163, no. 3 (2017):364-372,
https://doi.org/10.1099/mic.0.000429 . .