Comprehensive Analysis of <named-content content-type="genus-species">Aspergillus nidulans</named-content> PKA Phosphorylome Identifies a Novel Mode of CreA Regulation

Liliane F. C. Ribeiro; Cynthia Chelius; Karthik R. Boppidi; Nisha S. Naik; Simin Hossain; Jessica J. J. Ramsey; Jyothi Kumar; Lucas F. Ribeiro; Marc Ostermeier; Bao Tran; Young Ah Goo; Leandro J. de Assis; Mevlut Ulas; Ozgur Bayram; Gustavo H. Goldman; Stephen Lincoln; Ranjan Srivastava; Steven D. Harris; Mark R. Marten

doi:10.1128/mBio.02825-18

mBio (Apr 2019)

Comprehensive Analysis of <named-content content-type="genus-species">Aspergillus nidulans</named-content> PKA Phosphorylome Identifies a Novel Mode of CreA Regulation

Liliane F. C. Ribeiro,
Cynthia Chelius,
Karthik R. Boppidi,
Nisha S. Naik,
Simin Hossain,
Jessica J. J. Ramsey,
Jyothi Kumar,
Lucas F. Ribeiro,
Marc Ostermeier,
Bao Tran,
Young Ah Goo,
Leandro J. de Assis,
Mevlut Ulas,
Ozgur Bayram,
Gustavo H. Goldman,
Stephen Lincoln,
Ranjan Srivastava,
Steven D. Harris,
Mark R. Marten

Affiliations

Liliane F. C. Ribeiro: Department of Chemical, Biochemical and Environmental Engineering, University of Maryland, Baltimore County (UMBC), Baltimore, Maryland, USA
Cynthia Chelius: Department of Chemical, Biochemical and Environmental Engineering, University of Maryland, Baltimore County (UMBC), Baltimore, Maryland, USA
Karthik R. Boppidi: Department of Chemical, Biochemical and Environmental Engineering, University of Maryland, Baltimore County (UMBC), Baltimore, Maryland, USA
Nisha S. Naik: Department of Chemical, Biochemical and Environmental Engineering, University of Maryland, Baltimore County (UMBC), Baltimore, Maryland, USA
Simin Hossain: Department of Chemical, Biochemical and Environmental Engineering, University of Maryland, Baltimore County (UMBC), Baltimore, Maryland, USA
Jessica J. J. Ramsey: Department of Chemical, Biochemical and Environmental Engineering, University of Maryland, Baltimore County (UMBC), Baltimore, Maryland, USA
Jyothi Kumar: Center for Plant Science Innovation, University of Nebraska, Lincoln, Nebraska, USA
Lucas F. Ribeiro: Department of Chemical and Biomolecular Engineering, Johns Hopkins University, Baltimore, Maryland, USA
Marc Ostermeier: Department of Chemical and Biomolecular Engineering, Johns Hopkins University, Baltimore, Maryland, USA
Bao Tran: Mass Spectrometry Center, University of Maryland School of Pharmacy, Baltimore, Maryland, USA
Young Ah Goo: Mass Spectrometry Center, University of Maryland School of Pharmacy, Baltimore, Maryland, USA
Leandro J. de Assis: Departamento de Ciências Farmacêuticas, Faculdade de Ciências Farmacêuticas de Ribeirão Preto, Universidade de São Paulo, São Paulo, Brazil
Mevlut Ulas: Department of Biology, Maynooth University, Maynooth, County Kildare, Ireland
Ozgur Bayram: Department of Biology, Maynooth University, Maynooth, County Kildare, Ireland
Gustavo H. Goldman: Faculdade de Ciências Farmacêuticas de Ribeirão Preto, Universidade de São Paulo, CEP 14040-903, São Paulo, Brazil
Stephen Lincoln: Chemical & Biomolecular Engineering Department, University of Connecticut, Storrs, Connecticut, USA
Ranjan Srivastava: Chemical & Biomolecular Engineering Department, University of Connecticut, Storrs, Connecticut, USA
Steven D. Harris: Center for Plant Science Innovation, University of Nebraska, Lincoln, Nebraska, USA
Mark R. Marten: Department of Chemical, Biochemical and Environmental Engineering, University of Maryland, Baltimore County (UMBC), Baltimore, Maryland, USA

DOI: https://doi.org/10.1128/mBio.02825-18
Journal volume & issue: Vol. 10, no. 2

Abstract

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ABSTRACT 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.

Published in mBio

ISSN: 2161-2129 (Print); 2150-7511 (Online)
Publisher: American Society for Microbiology
Country of publisher: United States
LCC subjects: Science: Microbiology
Website: https://journals.asm.org/journal/mbio

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