Fusarium verticillioides NAT1 ( FDB2 ) N ‐malonyltransferase is structurally, functionally and phylogenetically distinct from its N ‐acetyltransferase ( NAT ) homologues

Karagianni, Eleni‐Pavlina, Kontomina, Evanthia, Lowe, Edward D., Athanasopoulos, Konstantinos, Papanikolaou, Georgia, Garefalaki, Vasiliki, Kotseli, Varvara, Zaliou, Sofia, Grimaud, Tom, Arvaniti, Konstantina, Tsatiri, Maria‐Aggeliki, Fakis, Giannoulis, Glenn, Anthony E., Roversi, Pietro, Abuhammad, Areej, Ryan, Ali, Sim, Robert B., Sim, Edith and Boukouvala, Sotiria (2022) Fusarium verticillioides NAT1 ( FDB2 ) N ‐malonyltransferase is structurally, functionally and phylogenetically distinct from its N ‐acetyltransferase ( NAT ) homologues. FEBS Journal. ISSN 1742-464X (In Press)

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Official URL: https://doi.org/10.1111/febs.16642


Fusarium endophytes damage cereal crops and contaminate produce with mycotoxins. Those fungi overcome the main chemical defence of host via detoxification by a malonyl-CoA-dependent enzyme homologous to xenobiotic metabolizing arylamine N-acetyltransferase (NAT). In Fusarium verticillioides (teleomorph Gibberella moniliformis, GIBMO), this N-malonyltransferase activity is attributed to (GIBMO)NAT1, and the fungus has two additional isoenzymes, (GIBMO)NAT3 (N-acetyltransferase) and (GIBMO)NAT2 (unknown function). We present the crystallographic structure of (GIBMO)NAT1, also modelling other fungal NAT homologues. Monomeric (GIBMO)NAT1 is distinctive, with access to the catalytic core through two “tunnel-like” entries separated by a “bridge-like” helix. In the quaternary arrangement, (GIBMO)NAT1 monomers interact in pairs along an extensive interface whereby one entry of each monomer is covered by the N-terminus of the other monomer. Although monomeric (GIBMO)NAT1 apparently accommodates acetyl-CoA better than malonyl-CoA, dimerization changes the active site to allow malonyl-CoA to reach the catalytic triad (Cys110, His158 and Asp173) via the single uncovered entry, and anchor its terminal carboxyl-group via hydrogen bonds to Arg109, Asn157 and Thr261. Lacking a terminal carboxyl-group, acetyl-CoA cannot form such stabilizing interactions, while longer acyl-CoAs enter the active site but cannot reach catalytic Cys. Other NAT isoenzymes lack such structural features, with (GIBMO)NAT3 resembling bacterial NATs and (GIBMO)NAT2 adopting a structure intermediate between (GIBMO)NAT1 and (GIBMO)NAT3. Biochemical assays confirmed differential donor substrate preference of (GIBMO)NAT isoenzymes, with phylogenetic analysis demonstrating evolutionary separation. Given the role of (GIBMO)NAT1 in enhancing Fusarium pathogenicity, unravelling the structure and function of this enzyme may benefit research into more targeted strategies for pathogen control.

Item Type: Article
Additional Information: Funding information: The authors would thank Nicole Crenshaw and Britton Glenn for their technical assistance with PCR and sequencing of the gene from different species. We also acknowledge the support of international mobility programmes providing opportunities for collaboration, as follows: (a) Democritus University of Thrace internship to EK to visit Oxford (UK) in 2011; (b) European Union FP7 “Capacities” Coordination & Support Action REGPOT‐2008‐1 programme (acronym BioStrength) grant to GF and SB to visit Oxford (UK), and to AEG to visit Alexandroupolis (Greece), in 2011; (c) Fulbright‐Schuman research scholarship to SB to visit Athens (GA, USA), in 2012; (d) Democritus University of Thrace Leonardo da Vinci scholarship to SZ to visit Kingston‐upon‐Thames (UK) in 2014; (e) Erasmus+ International Credit Mobility grant to SB to visit Athens (GA, USA) in 2018; (f) University of Angers internship to TG in 2021 to work with the Greek team. Additionally, PR was the recipient of a LISCB Wellcome Trust ISSF award, grant reference 204801/Z/16/Z, and of a Wellcome Trust Seed Award in Science, grant reference 214090/Z/18/Z; AA was the recipient of a University of Jordan Ph.D. scholarship, while at Oxford; EK was the recipient of a Ph.D. scholarship (2016–2019) co‐financed by Greece and the European Union (European Social Fund‐ESF) through Operational Programme “Human Resources Development, Education and Lifelong Learning” in the context of project “Strengthening Human Resources Research Potential via Doctorate Research” (MIS‐5000432), implemented by the State Scholarships Foundation (ΙΚΥ). The funding sources had no involvement in the conduct of the research, the preparation of the article or the decision to submit the article for publication.
Uncontrolled Keywords: Fusarium verticillioides; Gibberella moniliformis; N-malonyltransferase; N-acetyltransferase; protein crystal structure
Subjects: C400 Genetics
C700 Molecular Biology, Biophysics and Biochemistry
Department: Faculties > Health and Life Sciences > Applied Sciences
Depositing User: Elena Carlaw
Date Deposited: 29 Nov 2022 16:38
Last Modified: 29 Nov 2022 16:45
URI: https://nrl.northumbria.ac.uk/id/eprint/50765

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