TY - JOUR
T1 - Comprehensive analysis of aspergillus nidulans PKA phosphorylome identifies a novel mode of CreA regulation
AU - Ribeiro, Liliane F.C.
AU - Chelius, Cynthia
AU - Boppidi, Karthik R.
AU - Naik, Nisha S.
AU - Hossain, Simin
AU - Ramsey, Jessica J.J.
AU - Kumar, Jyothi
AU - Ribeiro, Lucas F.
AU - Ostermeier, Marc
AU - Tran, Bao
AU - Goo, Young Ah
AU - Assis, Leandro J.
AU - Ulas, Mevlut
AU - Bayram, Ozgur
AU - Goldman, Gustavo H.
AU - Lincoln, Stephen
AU - Srivastava, Ranjan
AU - Harris, Steven D.
AU - Martena, Mark R.
N1 - Funding Information:
This material is based upon work supported, in part, by the National Science Foundation under grants 1517309 and 1601935. Additional financial support was provided by the Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) (2016/20358-5 and 2016/18827-7) and Conselho Nacional de Desenvolvimento Cientí-fico e Tecnológico (CNPq), both in Brazil.
Funding Information:
Additional financial support was provided by the Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP), (2016/20358-5 and 2016/18827-7) and Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq), both from Brazil, and with the support of the Technical University of Munich – Institute for Advanced Study, funded by the German Excellence Initiative (G.H.G.).
Publisher Copyright:
© 2019 Ribeiro et al.
PY - 2019/3/1
Y1 - 2019/3/1
N2 - In filamentous fungi, an important kinase responsible for adaptation to changes in available nutrients is cyclic AMP (cAMP)-dependent protein kinase (protein kinase A [PKA]). This kinase has been well characterized at a molecular level, but its systemic action and direct/indirect targets are generally not well understood in filamentous fungi. In this work, we used a pkaA deletion strain (ΔpkaA) to identify Aspergillus nidulans proteins for which phosphorylation is dependent (either directly or indirectly) on PKA. A combination of phosphoproteomic and transcriptomic analyses revealed both direct and indirect targets of PKA and provided a global perspective on its function. One of these targets was the transcription factor CreA, the main repressor responsible for carbon catabolite repression (CCR). In the ΔpkaA strain, we identified a previously unreported phosphosite in CreA, S319, which (based on motif analysis) appears to be a direct target of Stk22 kinase (AN5728). Upon replacement of CreA S319 with an alanine (i.e., phosphonull mutant), the dynamics of CreA import to the nucleus are affected. Collectively, this work provides a global overview of PKA function while also providing novel insight regarding significance of a specific PKA-mediated phosphorylation event. IMPORTANCE The cyclic AMP (cAMP)-dependent protein kinase A (PKA) signaling pathway is well conserved across eukaryotes, and previous work has shown that it plays an important role in regulating development, growth, and virulence in a number of fungi. PKA is activated in response to extracellular nutrients and acts to regulate metabolism and growth. While a number of components in the PKA pathway have been defined in filamentous fungi, current understanding does not provide a global perspective on PKA function. Thus, this work is significant in that it comprehensively identifies proteins and functional pathways regulated by PKA in a model filamentous fungus. This information enhances our understanding of PKA action and may provide information on how to manipulate it for specific purposes.
AB - In filamentous fungi, an important kinase responsible for adaptation to changes in available nutrients is cyclic AMP (cAMP)-dependent protein kinase (protein kinase A [PKA]). This kinase has been well characterized at a molecular level, but its systemic action and direct/indirect targets are generally not well understood in filamentous fungi. In this work, we used a pkaA deletion strain (ΔpkaA) to identify Aspergillus nidulans proteins for which phosphorylation is dependent (either directly or indirectly) on PKA. A combination of phosphoproteomic and transcriptomic analyses revealed both direct and indirect targets of PKA and provided a global perspective on its function. One of these targets was the transcription factor CreA, the main repressor responsible for carbon catabolite repression (CCR). In the ΔpkaA strain, we identified a previously unreported phosphosite in CreA, S319, which (based on motif analysis) appears to be a direct target of Stk22 kinase (AN5728). Upon replacement of CreA S319 with an alanine (i.e., phosphonull mutant), the dynamics of CreA import to the nucleus are affected. Collectively, this work provides a global overview of PKA function while also providing novel insight regarding significance of a specific PKA-mediated phosphorylation event. IMPORTANCE The cyclic AMP (cAMP)-dependent protein kinase A (PKA) signaling pathway is well conserved across eukaryotes, and previous work has shown that it plays an important role in regulating development, growth, and virulence in a number of fungi. PKA is activated in response to extracellular nutrients and acts to regulate metabolism and growth. While a number of components in the PKA pathway have been defined in filamentous fungi, current understanding does not provide a global perspective on PKA function. Thus, this work is significant in that it comprehensively identifies proteins and functional pathways regulated by PKA in a model filamentous fungus. This information enhances our understanding of PKA action and may provide information on how to manipulate it for specific purposes.
KW - CreA
KW - Phosphoproteomic
KW - Phosphosite
KW - Signal transduction
KW - Transcriptomic
UR - http://www.scopus.com/inward/record.url?scp=85065464463&partnerID=8YFLogxK
U2 - 10.1128/mBio.02825-18
DO - 10.1128/mBio.02825-18
M3 - Article
C2 - 31040248
AN - SCOPUS:85065464463
VL - 10
JO - mBio
JF - mBio
SN - 2161-2129
IS - 2
M1 - e02825-18
ER -