Regulation of inorganic sulfate activation in filamentous fungi. Allosteric inhibition of ATP sulfurylase by 3'-phosphoadenosine-5'-phosphosulfate.

Sulfur metabolism has been implicated in the virulence, antibiotic resistance and anti-oxidant defence of Mycobacterium tuberculosis. Despite its human disease relevance, sulfur metabolism in mycobacteria has not yet been fully characterized. ATP sulfurylase catalyses the synthesis of activated sulfate (adenosine 5′-phosphosulfate, APS), the first step in the reductive assimilation of sulfate. Expression of the M. tuberculosis cysD gene, predicted to encode the adenylyl-transferase subunit of ATP sulfurylase, is upregulated by the bacilli inside its preferred host, the macrophage. This study demonstrates that cysD and cysNC orthologues exist in M. tuberculosis and constitute an operon whose expression is induced by sulfur limitation and repressed by the presence of cysteine, a major end-product of sulfur assimilation. The cysDNC genes are also induced upon exposure to oxidative stress, suggesting regulation of sulfur assimilation by M. tuberculosis in response to toxic oxidants. To ensure that the cysDNC operon encoded the activities predicted by its primary sequence, and to begin to characterize the products of the operon, they were expressed in Escherichia coli, purified to homogeneity, and tested for their catalytic activities. The CysD and CysNC proteins were shown to form a multifunctional enzyme complex that exhibits the three linked catalytic activities that constitute the sulfate activation pathway.

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Import of Entamoeba histolytica Mitosomal ATP Sulfurylase Relies on Internal Targeting Sequences

Microorganisms ◽

10.3390/microorganisms8081229 ◽

2020 ◽

Vol 8 (8) ◽

pp. 1229

Author(s):

Herbert J. Santos ◽

Yoko Chiba ◽

Takashi Makiuchi ◽

Saki Arakawa ◽

Yoshitaka Murakami ◽

...

Keyword(s):

Entamoeba Histolytica ◽

Intestinal Parasite ◽

Subcellular Fractionation ◽

Immunofluorescence Assay ◽

Matrix Proteins ◽

Desulfovibrio Vulgaris ◽

Chimeric Proteins ◽

Atp Sulfurylase ◽

Sulfate Activation

Mitochondrial matrix proteins synthesized in the cytosol often contain amino (N)-terminal targeting sequences (NTSs), or alternately internal targeting sequences (ITSs), which enable them to be properly translocated to the organelle. Such sequences are also required for proteins targeted to mitochondrion-related organelles (MROs) that are present in a few species of anaerobic eukaryotes. Similar to other MROs, the mitosomes of the human intestinal parasite Entamoeba histolytica are highly degenerate, because a majority of the components involved in various processes occurring in the canonical mitochondria are either missing or modified. As of yet, sulfate activation continues to be the only identified role of the relic mitochondria of Entamoeba. Mitosomes influence the parasitic nature of E. histolytica, as the downstream cytosolic products of sulfate activation have been reported to be essential in proliferation and encystation. Here, we investigated the position of the targeting sequence of one of the mitosomal matrix enzymes involved in the sulfate activation pathway, ATP sulfurylase (AS). We confirmed by immunofluorescence assay and subcellular fractionation that hemagluttinin (HA)-tagged EhAS was targeted to mitosomes. However, its ortholog in the δ-proteobacterium Desulfovibrio vulgaris, expressed as DvAS-HA in amoebic trophozoites, indicated cytosolic localization, suggesting a lack of recognizable mitosome targeting sequence in this protein. By expressing chimeric proteins containing swapped sequences between EhAS and DvAS in amoebic cells, we identified the ITSs responsible for mitosome targeting of EhAS. This observation is similar to other parasitic protozoans that harbor MROs, suggesting a convergent feature among various MROs in favoring ITS for the recognition and translocation of targeted proteins.

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Allosteric inhibition via R-state destabilization in ATP sulfurylase from Penicillium chrysogenum

Natural Structural Biology ◽

10.1038/nsb868 ◽

2002 ◽

Vol 9 (12) ◽

pp. 945-949 ◽

Cited By ~ 18

Author(s):

Ian J. MacRae ◽

Irwin H. Segel ◽

Andrew J. Fisher

Keyword(s):

Penicillium Chrysogenum ◽

Atp Sulfurylase ◽

Allosteric Inhibition

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ATP Sulfurylase from Filamentous Fungi: Which Sulfonucleotide Is the True Allosteric Effector?

Archives of Biochemistry and Biophysics ◽

10.1006/abbi.1996.9751 ◽

1997 ◽

Vol 337 (1) ◽

pp. 17-26 ◽

Cited By ~ 11

Author(s):

Ian MacRae ◽

Irwin H. Segel

Keyword(s):

Filamentous Fungi ◽

Atp Sulfurylase ◽

Allosteric Effector

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Modulation of the complex regulatory network for methionine biosynthesis in fungi

Genetics ◽

10.1093/genetics/iyaa049 ◽

2021 ◽

Vol 217 (2) ◽

Cited By ~ 1

Author(s):

Manjari Shrivastava ◽

Jinrong Feng ◽

Mark Coles ◽

Benjamin Clark ◽

Amjad Islam ◽

...

Keyword(s):

Filamentous Fungi ◽

Fungal Pathogen ◽

Biosynthetic Pathway ◽

Protein A ◽

Central Metabolism ◽

Methionine Biosynthesis ◽

Central Elements ◽

Sulfur Containing ◽

Complex Regulatory Network ◽

Inorganic Sulfate

Abstract The assimilation of inorganic sulfate and the synthesis of the sulfur-containing amino acids methionine and cysteine is mediated by a multibranched biosynthetic pathway. We have investigated this circuitry in the fungal pathogen Candida albicans, which is phylogenetically intermediate between the filamentous fungi and Saccharomyces cerevisiae. In S. cerevisiae, this pathway is regulated by a collection of five transcription factors (Met4, Cbf1, Met28, and Met31/Met32), while in the filamentous fungi the pathway is controlled by a single Met4-like factor. We found that in C. albicans, the Met4 ortholog is also a core regulator of methionine biosynthesis, where it functions together with Cbf1. While C. albicans encodes this Met4 protein, a Met4 paralog designated Met28 (Orf19.7046), and a Met31 protein, deletion, and activation constructs suggest that of these proteins only Met4 is actually involved in the regulation of methionine biosynthesis. Both Met28 and Met31 are linked to other functions; Met28 appears essential, and Met32 appears implicated in the regulation of genes of central metabolism. Therefore, while S. cerevisiae and C. albicans share Cbf1 and Met4 as central elements of the methionine biosynthesis control, the other proteins that make up the circuit in S. cerevisiae are not members of the C. albicans control network, and so the S. cerevisiae circuit likely represents a recently evolved arrangement.

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Mechanism of Sulfate Activation Catalyzed by ATP Sulfurylase - Magnesium Inhibits the Activity

Computational and Structural Biotechnology Journal ◽

10.1016/j.csbj.2019.06.016 ◽

2019 ◽

Vol 17 ◽

pp. 770-784 ◽

Cited By ~ 2

Author(s):

Anna Wójcik-Augustyn ◽

A. Johannes Johansson ◽

Tomasz Borowski

Keyword(s):

Atp Sulfurylase ◽

Sulfate Activation

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Crystal structure of ATP sulfurylase from Saccharomyces cerevisiae, a key enzyme in sulfate activation

The EMBO Journal ◽

10.1093/emboj/20.3.316 ◽

2001 ◽

Vol 20 (3) ◽

pp. 316-329 ◽

Cited By ~ 69

Author(s):

T. C. Ullrich

Keyword(s):

Crystal Structure ◽

Saccharomyces Cerevisiae ◽

Atp Sulfurylase ◽

Key Enzyme ◽

Sulfate Activation

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Rhizobium meliloti genes involved in sulfate activation: the two copies of nodPQ and a new locus, saa.

Genetics ◽

10.1093/genetics/132.4.899 ◽

1992 ◽

Vol 132 (4) ◽

pp. 899-909 ◽

Cited By ~ 5

Author(s):

J S Schwedock ◽

S R Long

Keyword(s):

Host Plants ◽

Rhizobium Meliloti ◽

Atp Sulfurylase ◽

Plant Host ◽

Nod Factor ◽

Aps Kinase ◽

Specific Host ◽

Cysteine Synthesis ◽

Sulfate Activation ◽

Leguminous Host

Abstract The nitrogen-fixing symbiont Rhizobium meliloti establishes nodules on leguminous host plants. Nodulation (nod) genes used for this process are located in a cluster on the pSym-a megaplasmid of R. meliloti. These genes include nodP and nodQ (here termed nodPQ), which encode ATP sulfurylase and APS kinase, enzymes that catalyze the conversion of ATP and SO(4)2- into the activated sulfate form 3'-phosphoadenosine 5'-phosphosulfate (PAPS), an intermediate in cysteine synthesis. In Rhizobium, PAPS is also a precursor for sulfated and N-acylated oligosaccharide Nod-factor signals that cause symbiotic responses on specific host plants such as alfalfa. We previously found a highly conserved second copy of nodPQ in R. meliloti. We report here the mapping and cloning of this second copy, and its location on the second megaplasmid, pSym-b. The function of nodP2Q2 is equivalent to that of nodP1Q1 in complementation tests of R. meliloti and Escherichia coli mutants in ATP sulfurylase and adenosine 5'-phosphosulfate (APS) kinase. Mutations in nodP2Q2 do not have as severe an effect on symbiosis or plant host range as do those in nodP1Q1, however, possibly reflecting differences in expression and/or channeling of metabolites to specific enzymes involved in sulfate transfer. Strains mutated or deleted for both copies of nodQ are severely defective in symbiotic phenotypes, but remain prototrophic. This suggests the existence in R. meliloti of a third locus for ATP sulfurylase and APS kinase activities. We have found a new locus saa (sulfur amino acid), which may also encode these activities.

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