Fungal-induced protein hyperacetylation in maize identified by acetylome profiling

Lysine acetylation is a key posttranslational modification that regulates diverse proteins involved in a range of biological processes. The role of histone acetylation in plant defense is well established, and it is known that pathogen effector proteins encoding acetyltransferases can directly acetylate host proteins to alter immunity. However, it is unclear whether endogenous plant enzymes can modulate protein acetylation during an immune response. Here, we investigate how the effector molecule HC-toxin (HCT), a histone deacetylase inhibitor produced by the fungal pathogen Cochliobolus carbonum race 1, promotes virulence in maize through altering protein acetylation. Using mass spectrometry, we globally quantified the abundance of 3,636 proteins and the levels of acetylation at 2,791 sites in maize plants treated with HCT as well as HCT-deficient or HCT-producing strains of C. carbonum. Analyses of these data demonstrate that acetylation is a widespread posttranslational modification impacting proteins encoded by many intensively studied maize genes. Furthermore, the application of exogenous HCT enabled us to show that the activity of plant-encoded enzymes (histone deacetylases) can be modulated to alter acetylation of nonhistone proteins during an immune response. Collectively, these results provide a resource for further mechanistic studies examining the regulation of protein function by reversible acetylation and offer insight into the complex immune response triggered by virulent C. carbonum.

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Fungal Induced Protein Hyperacetylation Identified by Acetylome Profiling

10.1101/057174 ◽

2016 ◽

Author(s):

Justin W Walley ◽

Zhouxin Shen ◽

Maxwell R. McReynolds ◽

Steven P. Briggs

Keyword(s):

Immune Response ◽

Protein Function ◽

Effector Proteins ◽

Protein Acetylation ◽

Post Translational Modification ◽

Cochliobolus Carbonum ◽

Pathogen Virulence ◽

Pathogen Effector ◽

Race 1 ◽

Hc Toxin

ABSTRACTLysine acetylation is a key post-translational modification that regulates diverse proteins involved in a range of biological processes. The role of histone acetylation in plant defense is well established and it is known that pathogen effector proteins encoding acetyltransferses can directly acetylate host proteins to alter immunity. However, it is unclear whether endogenous plant enzymes can modulate protein acetylation during an immune response. Here we investigate how the effector molecule HC-toxin, a histone deacetylase inhibitor, produced by Cochliobolus carbonum race 1 promotes pathogen virulence in maize through altering protein acetylation. Using mass spectrometry we globally quantified the abundance of 3,636 proteins and the levels of acetylation at 2,791 sites in maize plants treated with HC-toxin as well as HC-toxin deficient or producing strains of C. carbonum. Analyses of these data demonstrate that acetylation is a widespread post-translational modification impacting proteins encoded by many intensively studied maize genes. Furthermore, the application of exogenous HC-toxin enabled us to show that the activity of plant-encoded enzymes can be modulated to alter acetylation of non-histone proteins during an immune response. Collectively, these results provide a resource for further mechanistic studies examining the regulation of protein function and offer insight into the complex immune response triggered by virulent C. carbonum.

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Inhibition of maize histone deacetylases by HC toxin, the host-selective toxin of Cochliobolus carbonum.

The Plant Cell ◽

10.1105/tpc.7.11.1941 ◽

1995 ◽

Vol 7 (11) ◽

pp. 1941-1950 ◽

Cited By ~ 117

Author(s):

G Brosch ◽

R Ransom ◽

T Lechner ◽

J D Walton ◽

P Loidl

Keyword(s):

Histone Deacetylases ◽

Cochliobolus Carbonum ◽

Hc Toxin

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Modulation of Acetylation: Creating a Pro-survival and Anti-Inflammatory Phenotype in Lethal Hemorrhagic and Septic Shock

Journal of Biomedicine and Biotechnology ◽

10.1155/2011/523481 ◽

2011 ◽

Vol 2011 ◽

pp. 1-15 ◽

Cited By ~ 23

Author(s):

Yongqing Li ◽

Hasan B. Alam

Keyword(s):

Septic Shock ◽

Histone Deacetylases ◽

Histone Deacetylase Inhibitors ◽

Clinical Applications ◽

Critical Conditions ◽

Protein Acetylation ◽

Nonhistone Proteins ◽

Recent Developments ◽

Inflammatory Phenotype ◽

Anti Inflammatory

Histone deacetylases (HDACs) play a key role in homeostasis of protein acetylation in histone and nonhistone proteins and in regulating fundamental cellular activities. In this paper we review and discuss intriguing recent developments in the use of histone deacetylase inhibitors (HDACIs) to combat some critical conditions in an animal model of hemorrhagic and septic shock. HDACIs have neuroprotective, cardioprotective, renal-protective, and anti-inflammatory properties; survival improvements have been significantly shown in these models. We discuss the targets and mechanisms underlying these effects of HDACIs and comment on the potential new clinical applications for these agents in the future. This paper highlights the emerging roles of HDACIs as acetylation modulators in models of hemorrhagic and septic shock and explains some contradictions encountered in previous studies.

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Inhibition of Maize Histone Deacetylases by HC Toxin, the Host-Selective Toxin of Cochliobolus carbonum

The Plant Cell ◽

10.2307/3870201 ◽

1995 ◽

Vol 7 (11) ◽

pp. 1941 ◽

Cited By ~ 5

Author(s):

Gerald Brosch ◽

Richard Ransom ◽

Thomas Lechner ◽

Jonathan D. Walton ◽

Peter Loidl

Keyword(s):

Histone Deacetylases ◽

Cochliobolus Carbonum ◽

Hc Toxin

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Cross-reactivity of a rice NLR immune receptor to distinct effectors from the blast pathogen leads to partial disease resistance

10.1101/530675 ◽

2019 ◽

Cited By ~ 2

Author(s):

Freya A. Varden ◽

Hiromasa Saitoh ◽

Kae Yoshino ◽

Marina Franceschetti ◽

Sophien Kamoun ◽

...

Keyword(s):

Immune Response ◽

Disease Resistance ◽

Effector Proteins ◽

In Planta ◽

Immune Receptor ◽

Immune Receptors ◽

Binding Interface ◽

Pathogen Effector ◽

Integrated Domains

ABSTRACTUnconventional integrated domains in plant intracellular immune receptors (NLRs) can directly bind translocated pathogen effector proteins to initiate an immune response. The rice immune receptor pairs Pik-1/Pik-2 and RGA5/RGA4 both use integrated heavy metal-associated (HMA) domains to bind the Magnaporthe oryzae effectors AVR-Pik and AVR-Pia, respectively. These effectors both belong to the MAX effector family and share a core structural fold, despite being divergent in sequence. How integrated domains maintain specificity of recognition, even for structurally similar effectors, has implications for understanding plant immune receptor evolution and function. Here we show that the rice NLR pair Pikp-1/Pikp-2 triggers an immune response leading to partial disease resistance towards the “mismatched” effector AVR-Pia in planta, and that the Pikp-HMA domain binds AVR-Pia in vitro. The HMA domain from another Pik-1 allele, Pikm, is unable to bind AVR-Pia, and does not trigger a response in plants. The crystal structure of Pikp-HMA bound to AVR-Pia reveals a different binding interface compared to AVR-Pik effectors, suggesting plasticity in integrated domain/effector interactions. This work shows how a single NLR can bait multiple pathogen effectors via an integrated domain, and may enable engineering immune receptors with extended disease resistance profiles.

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A Fatty Acid Synthase Gene in Cochliobolus carbonum Required for Production of HC-Toxin, Cyclo(d-Prolyl-l-Alanyl- d-Alanyl- l-2-Amino-9,10-Epoxi-8-Oxodecanoyl)

Molecular Plant-Microbe Interactions ◽

10.1094/mpmi.1997.10.2.207 ◽

1997 ◽

Vol 10 (2) ◽

pp. 207-214 ◽

Cited By ~ 47

Author(s):

Joong-Hoon Ahn ◽

Jonathan D. Walton

Keyword(s):

Fatty Acid ◽

Fatty Acid Synthase ◽

Cyclic Peptide ◽

Decanoic Acid ◽

Toxin Production ◽

Gene Encoding ◽

Cochliobolus Carbonum ◽

Primary Amino ◽

Hc Toxin

The fungal maize pathogen Cochliobolus carbonum produces a phytotoxic and cytostatic cyclic peptide, HC-toxin, of structure cyclo(D-prolyl-L-alanyl-D-alanyl-L-Aeo), in which Aeo stands for 2-amino-9,10-epoxi-8-oxodecanoic acid. Here we report the isolation of a gene, TOXC, that is present only in HC-toxin-producing (Tox2+) fungal strains. TOXC is present in most Tox2+ strains in three functional copies, all of which are on the same chromosome as the gene encoding HC-toxin synthetase. When all copies of TOXC are mutated by targeted gene disruption, the fungus grows and sporulates normally in vitro but no longer makes HC-toxin and is not pathogenic, indicating that TOXC has a specific role in HC-toxin production and hence virulence. The TOXC mRNA is 6.5 kb and the predicted product has 2,080 amino acids and a molecular weight of 233,000. The primary amino acid sequence is highly similar (45 to 47% identity) to the β subunit of fatty acid synthase from several lower eukaryotes, and contains, in the same order as in other β subunits, domains predicted to encode acetyl transferase, enoyl reductase, dehydratase, and malonyl-palmityl transferase. The most plausible function of TOXC is to contribute to the synthesis of the decanoic acid backbone of Aeo.

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Cloning and Mapping of a Putative Barley NADPH-Dependent HC-Toxin Reductase

Molecular Plant-Microbe Interactions ◽

10.1094/mpmi.1997.10.2.234 ◽

1997 ◽

Vol 10 (2) ◽

pp. 234-239 ◽

Cited By ~ 18

Author(s):

F. Han ◽

A. Kleinhofs ◽

A. Kilian ◽

S. E. Ullrich

Keyword(s):

Plant Species ◽

Chromosome 9 ◽

Immature Embryos ◽

Chromosome 1 ◽

Grass Species ◽

Cochliobolus Carbonum ◽

Wide Range ◽

Young Leaves ◽

High Conservation ◽

Hc Toxin

The NADPH-dependent HC-toxin reductase (HCTR), encoded by Hm1 in maize, inactivates HC-toxin produced by the fungus Cochliobolus carbonum, and thus confers resistance to the pathogen. The fact that C. carbonum only infects maize (Zea mays) and is the only species known to produce HC-toxin raises the question: What are the biological functions of HCTR in other plant species? An HCTR-like enzyme may function to detoxify toxins produced by pathogens which infect other plant species (R. B. Meeley, G. S. Johal, S. E. Briggs, and J. D. Walton, Plant Cell, 4:71–77, 1992). Hm1 homolog in rice (Y. Hihara, M. Umeda, C. Hara, Q. Liu, S. Aotsuka, K. Toriyama, and H. Uchimiya, unpublished) and HCTR activity in barley, wheat, oats and sorghum have been reported (R. B. Meeley and J. D. Walton, Plant Physiol. 97:1080–1086, 1993). To investigate the sequence conservation of Hm1 and HCTR in barley and the possible relationship of barley Hm1 homolog to the known disease resistance genes, we cloned and mapped a barley (Hordeum vulgare) Hm1-like gene. A putative full-length cDNA clone, Bhm1-18, was isolated from a cDNA library consisting of mRNA from young leaves, inflorescences, and immature embryos. This 1,297-bp clone encodes 363 amino acids which show great similarity (81.6%) with the amino acid sequence of HM1 in maize. Two loci were mapped to barley molecular marker linkage maps with Bhm1-18 as the probe; locus A (Bhm1A) on the long arm of chromosome 1, and locus B (Bhm1B) on the short arm of chromosome 1 which is syntenic to maize chromosome 9 containing the Hm2 locus. The Bhm1-18 probe hybridized strongly to a Southern blot of a wide range of grass species, indicating high conservation of HCTR at the DNA sequence level among grasses. The HCTR mRNA was detected in barley roots, leaves, inflorescences, and immature embryos. The conservation of the HCTR sequence, together with its expression in other plant species (R. B. Meeley and J. D. Walton, Plant Physiol. 97:1080–1086, 1993), suggests HCTR plays an important functional role in other plant species.

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Histone deacetylase 1 reduces NO production in endothelial cells via lysine deacetylation of NO synthase 3

AJP Heart and Circulatory Physiology ◽

10.1152/ajpheart.00243.2014 ◽

2014 ◽

Vol 307 (5) ◽

pp. H803-H809 ◽

Cited By ~ 15

Author(s):

Kelly A. Hyndman ◽

Dao H. Ho ◽

Martiana F. Sega ◽

Jennifer S. Pollock

Keyword(s):

Endothelial Cells ◽

Histone Deacetylases ◽

Production Control ◽

No Synthase ◽

No Production ◽

Lysine Acetylation ◽

Nonhistone Proteins ◽

Endothelin 1 ◽

Histone Deacetylase 1 ◽

Nitrite Production

The lysine acetylation state of nonhistone proteins may be regulated through histone deacetylases (HDACs). Evidence suggests that nitric oxide (NO) synthase 3 (NOS3; endothelial NOS) is posttranslationally lysine acetylated, leading to increased NO production in the endothelium. We tested the hypothesis that NOS3 is lysine acetylated and that upregulated HDAC1-mediated deacetylation leads to reduced NO production in endothelial cells. We determined that NOS3 is basally lysine acetylated in cultured bovine aortic endothelial cells (BAECs). In BAECs, HDAC1 is expressed in the nucleus and cytosol and forms a novel protein-protein interaction with NOS3. Overexpression of HDAC1 in BAECs resulted in a significant reduction in NOS3 lysine acetylation (control = 1.0 ± 0.1 and HDAC1 = 0.59 ± 0.08 arbitrary units, P < 0.01) and significantly blunted basal nitrite production (control 287.7 ± 29.1 and HDAC1 172.4 ± 31.7 pmol·mg−1·h−1, P < 0.05) as well as attenuating endothelin-1-stimulated nitrite production (control = 481.8 ± 50.3 and HDAC1 243.1 ± 48.2 pmol·mg−1·h−1, P < 0.05). While HDAC1 knockdown with small-interfering RNA resulted in no change in NOS3 acetylation level, yet increased basal nitrite production (730.6 ± 99.1 pmol·mg−1·h−1) and further exaggerated increases in endothelin-1 stimulated nitrite production (1276.9 ± 288.2 pmol·mg−1·h−1) was observed. Moreover, overexpression or knockdown of HDAC1 resulted in no significant effect on NOS3 protein expression or NOS3 phosphorylation sites T497, S635, or S1179. Thus these data indicate that upregulated HDAC1 decreases NOS3 activity, most likely through direct lysine deacetylation of NOS3. We propose that HDAC1-mediated deacetylation of NOS3 may represent a novel target for endothelial dysfunction.

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