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Homologues of xenobiotic metabolizing N-acetyltransferases in plant-associated fungi: Novel functions for an old enzyme family.

Karagianni EP, Kontomina E, Davis B, Kotseli B, Tsirka T, Garefalaki V, Sim E, Glenn AE, Boukouvala S - Sci Rep (2015)

Bottom Line: Plant-pathogenic fungi and their hosts engage in chemical warfare, attacking each other with toxic products of secondary metabolism and defending themselves via an arsenal of xenobiotic metabolizing enzymes.The third group generates minimal activity with acyl-CoA compounds that bind non-selectively to the proteins.We propose that fungal NAT isoenzymes may have evolved to perform diverse functions, potentially relevant to pathogen fitness, acetyl-CoA/propionyl-CoA intracellular balance and secondary metabolism.

View Article: PubMed Central - PubMed

Affiliation: Democritus University of Thrace, Department of Molecular Biology and Genetics, Alexandroupolis 68100, Greece.

ABSTRACT
Plant-pathogenic fungi and their hosts engage in chemical warfare, attacking each other with toxic products of secondary metabolism and defending themselves via an arsenal of xenobiotic metabolizing enzymes. One such enzyme is homologous to arylamine N-acetyltransferase (NAT) and has been identified in Fusarium infecting cereal plants as responsible for detoxification of host defence compound 2-benzoxazolinone. Here we investigate functional diversification of NAT enzymes in crop-compromising species of Fusarium and Aspergillus, identifying three groups of homologues: Isoenzymes of the first group are found in all species and catalyse reactions with acetyl-CoA or propionyl-CoA. The second group is restricted to the plant pathogens and is active with malonyl-CoA in Fusarium species infecting cereals. The third group generates minimal activity with acyl-CoA compounds that bind non-selectively to the proteins. We propose that fungal NAT isoenzymes may have evolved to perform diverse functions, potentially relevant to pathogen fitness, acetyl-CoA/propionyl-CoA intracellular balance and secondary metabolism.

No MeSH data available.


Related in: MedlinePlus

Effect of acyl-coenzyme A compounds on the Tm of fungal NAT proteins.Overview of Tm values determined by differential scanning fluorimetry for recombinant NAT isoenzymes of F. verticillioides (G. moniliformis-GIBM7), F. graminearum (G. zeae-GIBZE), F. oxysporum f.sp. lycopersici (FUSO4) and A. flavus (ASPFN), in the absence or presence of various acyl-CoAs. Two replicate experiments were performed, generating overlapping curves for which the average Tm (± standard deviation) is shown. The results for each set of experiments are presented in Supplementary Fig. S4.
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f2: Effect of acyl-coenzyme A compounds on the Tm of fungal NAT proteins.Overview of Tm values determined by differential scanning fluorimetry for recombinant NAT isoenzymes of F. verticillioides (G. moniliformis-GIBM7), F. graminearum (G. zeae-GIBZE), F. oxysporum f.sp. lycopersici (FUSO4) and A. flavus (ASPFN), in the absence or presence of various acyl-CoAs. Two replicate experiments were performed, generating overlapping curves for which the average Tm (± standard deviation) is shown. The results for each set of experiments are presented in Supplementary Fig. S4.

Mentions: Binding of acyl-CoAs to fungal NATs was investigated using differential scanning fluorimetry (DSF), a technique that detects changes in the thermal stability of recombinant proteins upon interaction with their specific ligands22. The method was applicable with all recombinant NAT proteins in our panel, except the NAT3 of F. oxysporum [(FUSO4)NAT3] and the NAT1 of A. nidulans [(EMENI)NAT1] which were recovered at less optimal levels of yield and purity (Supplementary Fig. S2). The Tm values of different fungal NAT homologues varied considerably from around 20 to over 55 °C, with the NAT3 homologues of F. verticillioides [(GIBM7)NAT3] and F. graminearum [(GIBZE)NAT3] at opposite extremes (Fig. 2 and Supplementary Fig. S4). The addition of acyl-CoAs increased the Tm values of NAT proteins, with a pattern that reflected exactly the results of our activity assays for group I N-acetyl/N-propionyltransferases and group II N-malonyltransferases. In the case of group I homologues [(GIBM7)NAT3, (GIBZE)NAT3 and (ASPFN)NAT2], the shift in Tm was always greater with acetyl-CoA, intermediate with n-propionyl-CoA and minimal with malonyl-CoA. Conversely, for group II N-malonyltransferases [(GIBM7)NAT1, (GIBZE)NAT1, (FUSO4)NAT1 and (ASPFN)NAT3], the increase in Tm was substantial only with malonyl-CoA (Fig. 2 and Supplementary Fig. S4).


Homologues of xenobiotic metabolizing N-acetyltransferases in plant-associated fungi: Novel functions for an old enzyme family.

Karagianni EP, Kontomina E, Davis B, Kotseli B, Tsirka T, Garefalaki V, Sim E, Glenn AE, Boukouvala S - Sci Rep (2015)

Effect of acyl-coenzyme A compounds on the Tm of fungal NAT proteins.Overview of Tm values determined by differential scanning fluorimetry for recombinant NAT isoenzymes of F. verticillioides (G. moniliformis-GIBM7), F. graminearum (G. zeae-GIBZE), F. oxysporum f.sp. lycopersici (FUSO4) and A. flavus (ASPFN), in the absence or presence of various acyl-CoAs. Two replicate experiments were performed, generating overlapping curves for which the average Tm (± standard deviation) is shown. The results for each set of experiments are presented in Supplementary Fig. S4.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4542470&req=5

f2: Effect of acyl-coenzyme A compounds on the Tm of fungal NAT proteins.Overview of Tm values determined by differential scanning fluorimetry for recombinant NAT isoenzymes of F. verticillioides (G. moniliformis-GIBM7), F. graminearum (G. zeae-GIBZE), F. oxysporum f.sp. lycopersici (FUSO4) and A. flavus (ASPFN), in the absence or presence of various acyl-CoAs. Two replicate experiments were performed, generating overlapping curves for which the average Tm (± standard deviation) is shown. The results for each set of experiments are presented in Supplementary Fig. S4.
Mentions: Binding of acyl-CoAs to fungal NATs was investigated using differential scanning fluorimetry (DSF), a technique that detects changes in the thermal stability of recombinant proteins upon interaction with their specific ligands22. The method was applicable with all recombinant NAT proteins in our panel, except the NAT3 of F. oxysporum [(FUSO4)NAT3] and the NAT1 of A. nidulans [(EMENI)NAT1] which were recovered at less optimal levels of yield and purity (Supplementary Fig. S2). The Tm values of different fungal NAT homologues varied considerably from around 20 to over 55 °C, with the NAT3 homologues of F. verticillioides [(GIBM7)NAT3] and F. graminearum [(GIBZE)NAT3] at opposite extremes (Fig. 2 and Supplementary Fig. S4). The addition of acyl-CoAs increased the Tm values of NAT proteins, with a pattern that reflected exactly the results of our activity assays for group I N-acetyl/N-propionyltransferases and group II N-malonyltransferases. In the case of group I homologues [(GIBM7)NAT3, (GIBZE)NAT3 and (ASPFN)NAT2], the shift in Tm was always greater with acetyl-CoA, intermediate with n-propionyl-CoA and minimal with malonyl-CoA. Conversely, for group II N-malonyltransferases [(GIBM7)NAT1, (GIBZE)NAT1, (FUSO4)NAT1 and (ASPFN)NAT3], the increase in Tm was substantial only with malonyl-CoA (Fig. 2 and Supplementary Fig. S4).

Bottom Line: Plant-pathogenic fungi and their hosts engage in chemical warfare, attacking each other with toxic products of secondary metabolism and defending themselves via an arsenal of xenobiotic metabolizing enzymes.The third group generates minimal activity with acyl-CoA compounds that bind non-selectively to the proteins.We propose that fungal NAT isoenzymes may have evolved to perform diverse functions, potentially relevant to pathogen fitness, acetyl-CoA/propionyl-CoA intracellular balance and secondary metabolism.

View Article: PubMed Central - PubMed

Affiliation: Democritus University of Thrace, Department of Molecular Biology and Genetics, Alexandroupolis 68100, Greece.

ABSTRACT
Plant-pathogenic fungi and their hosts engage in chemical warfare, attacking each other with toxic products of secondary metabolism and defending themselves via an arsenal of xenobiotic metabolizing enzymes. One such enzyme is homologous to arylamine N-acetyltransferase (NAT) and has been identified in Fusarium infecting cereal plants as responsible for detoxification of host defence compound 2-benzoxazolinone. Here we investigate functional diversification of NAT enzymes in crop-compromising species of Fusarium and Aspergillus, identifying three groups of homologues: Isoenzymes of the first group are found in all species and catalyse reactions with acetyl-CoA or propionyl-CoA. The second group is restricted to the plant pathogens and is active with malonyl-CoA in Fusarium species infecting cereals. The third group generates minimal activity with acyl-CoA compounds that bind non-selectively to the proteins. We propose that fungal NAT isoenzymes may have evolved to perform diverse functions, potentially relevant to pathogen fitness, acetyl-CoA/propionyl-CoA intracellular balance and secondary metabolism.

No MeSH data available.


Related in: MedlinePlus