A pyruvate‐stimulated adenylate cyclase has a sequence related to the <i>fes/fps</i> oncogenes and to eukaryotic cyclases

Peters, Elisabeth; Wilderspin, Andrew F.; Wood, Stephen P.; Zvelebil, Marketa; Sezer, Odile; Danchin, Antoine

doi:10.1111/j.1365-2958.1991.tb01890.x

Cited by 19 publications

(15 citation statements)

References 29 publications

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“…In E. coli, the molecule plays important roles in regulating carbon catabolite genes, such as the lactose and arabinose operons (11). Under conditions of glucose starvation, the intracellular concentration of cAMP becomes elevated, a general phenomenon found in many bacteria with the exception of Brevibacterium species, which apparently keeps the concentration of cAMP high when cells are grown on glucose minimal medium (22,27). Similarly, in C. glutamicum, the intracellular concentration of cAMP is kept low when cells are grown on acetate medium but high when they are grown on glucose medium, as evidenced in our growth experiment (Fig.…”

Section: Discussionmentioning

confidence: 99%

Identification and Characterization ofglxR, a Gene Involved in Regulation of Glyoxylate Bypass inCorynebacterium glutamicum

et al. 2004

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A corynebacterial clone, previously isolated by scoring repression of lacZYA fused to the aceB promoter of Corynebacterium glutamicum, was analyzed further. In the clone, an open reading frame designated glxR, consisting of 681 nucleotides and encoding a 24,957-Da protein, was found. The molecular mass of a native GlxR protein was estimated by gel filtration column chromatography to be 44,000 Da, suggesting that the protein formed dimers. The predicted amino acid sequence contained both cyclic AMP (cAMP)-and DNAbinding motifs and was homologous with the cAMP receptor protein family of proteins. The aceB-repressing activity of the glxR clone was markedly relieved in an Escherichia coli cya mutant, but the activity was restored in growth medium containing cAMP. In glucose medium, the intracellular cAMP concentration of C. glutamicum reached 22 nmol/mg of protein in the early exponential phase and then decreased further; but in acetate medium, the intracellular cAMP concentration was only 5 nmol/mg of protein and remained low throughout the growth phase. The expression of glxR was not affected by the carbon source. Binding of purified GlxR to the promoter region of aceB could be demonstrated only in the presence of cAMP. These data suggest that GlxR may form dimers which bind to the aceB promoter region in the presence of cAMP and repress the glyoxylate bypass genes.

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Section: Discussionmentioning

confidence: 99%

Identification and Characterization ofglxR, a Gene Involved in Regulation of Glyoxylate Bypass inCorynebacterium glutamicum

et al. 2004

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“…More than 30 different adenylyl cyclases and guanylyl cyclases share the common homologous cytoplasmic domains, designated as the cyclase domains (1). For the functional enzyme, the cyclase domains can work as the homo-oligomer, such as adenylyl cyclases from yeast, Trypanosome, Dictyostelium (ACG), Leishmania, Brevibacterium, and Rhizobium (23)(24)(25)(26)(27)(28)(29)(30). The cyclase domains can also exist as an obligatory heterodimer for enzymatic activity such as the C 1 and C 2 domains from type I to type IX adenylyl cyclases, Drosophila rutabaga, and Dictyostelium ACA, and the ␣ and ␤ subunits of soluble guanylyl cyclases (22,23,(31)(32)(33)(34)(35)(36)(37)(38)(39)(40)(41)(42)(43)(44)(45)(46).…”

Section: Isolation and Analysis Of Loss Of Function Ic 1 Iic 2 Mutants-mentioning

confidence: 99%

“…Third are the peripheral membrane proteins from yeast (Schizosaccharomyces pombe, Saccharomyces cerevisiae, and Saccharomyces klyuyveri) (26 -28). Fourth are the soluble enzymes from the bacteria, Brevibacterium liquefaciens (Bl AC) and Rhizobium meliloti (Rm AC) (29,30). 2 The abbreviations used are: G protein, guanine nucleotide binding regulatory protein; G s␣ , the ␣ subunit of the G protein that stimulates adenylyl cyclase; Fsk, forskolin; IC 1 protein, C 1 domain of type I adenylyl cyclase; IIC 2 protein, C 2 domain of type II adenylyl cyclase; GTP␥S, guanosine-5Ј-[␥-thio]triphosphate; aa, amino acid(s); 2Ј-D-3Ј-AMP, 2Јde-oxyadenosine 3Ј-monophosphate; kbp, kilobase pair; IPTG, isopropyl-1-thio-␤-D-galactopyranoside; LB, Luria broth; CMV, cytomegalovirus; IBMX, isobutylmethylxanthine.…”

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confidence: 99%

The Conserved Asparagine and Arginine Are Essential for Catalysis of Mammalian Adenylyl Cyclase

Yan

Huang

Shaw

et al. 1997

Journal of Biological Chemistry

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Mammalian adenylyl cyclases have two homologous cytoplasmic domains (C 1 and C 2 ), and both domains are required for the high enzymatic activity. Mutational and genetic analyses of type I and soluble adenylyl cyclases suggest that the C 2 domain is catalytically active and the C 1 domain is not; the role of the C 1 domain is to promote the catalytic activity of the C 2 domain. Two amino acid residues, Asn-1025 and Arg-1029 of type II adenylyl cyclase, are conserved among the C 2 domains, but not among the C 1 domains, of adenylyl cyclases with 12 putative transmembrane helices. Mutations at each amino acid residue alone result in a 30 -100-fold reduction in K cat of adenylyl cyclase. However, the same mutations do not affect the K m for ATP, the half-maximal concentration (EC 50 ) for the C 2 domain of type II adenylyl cyclase to associate with the C 1 domain of type I adenylyl cyclase and achieve maximal enzyme activity, or the EC 50 for forskolin to maximally activate enzyme activity with or without G s␣ . This indicates that the mutations at these two residues do not cause gross structural alteration. Thus, these two conserved amino acid residues appear to be crucial for catalysis, and their absence from the C 1 domains may account for its lack of catalytic activity. Mutations at both amino acid residues together result in a 3,000-fold reduction in K cat of adenylyl cyclase, suggesting that these two residues have additive effects in catalysis. A second site suppressor of the Asn-1025 to Ser mutant protein has been isolated. This suppressor has 17-fold higher activity than the mutant and has a Pro-1015 to Ser mutation.The activity of mammalian adenylyl cyclase, the enzyme that converts ATP to cAMP, is the key step in modulating intracellular cAMP concentration in response to extracellular stimulation by hormones, neurotransmitters, and odorants. Nine isoforms of mammalian adenylyl cyclases have been cloned to date, and they belong to a rapid expanding cyclase family that includes class III adenylyl cyclases 1 and guanylyl cyclases (1).All nine isoforms have a similar structure, including two intensely hydrophobic domains (M 1 and M 2 ) and two 40-kDa cytoplasmic domains (C 1 and C 2 ). The two cytoplasmic domains contain sequences (C 1a and C 2a ) that are homologous to each other and to class III adenylyl cyclases and guanylyl cyclases. Each isoform of adenylyl cyclase not only has distinct patterns of tissue expression but also has unique responses to extracellular and intracellular stimuli (1-3). In common, each adenylyl cyclase can be activated by the G protein 2 ␣ subunit, designated as G s␣ , and each can be inhibited by certain adenosine analogues (P-site inhibitors). However, there are several subtype-specific regulators, including forskolin, G protein ␤␥ subunits, the ␣ subunit of G i , G o , and G z , Ca 2ϩ ion, Ca 2ϩ -calmodulin, Ca 2ϩ -calcineurin, cAMP-dependent protein kinase, and PKC (for review, see Refs. 1-3). Certain ones of these regulators (e.g. ␤␥) can be either stimulatory or inhibitory...

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“…Earlier, AC genes of this subtype were identified by complementation cloning in Brevibacterium liquefaciens and Streptomyces (5,6). Those genes code for modular proteins with the CHD located C-terminally.…”

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Adenylyl Cyclase Rv1264 from Mycobacterium tuberculosis Has an Autoinhibitory N-terminal Domain

Linder

Schultz

2002

Journal of Biological Chemistry

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Mycobacterium tuberculosis contains 15 class III adenylyl cyclase genes. The gene Rv1264 is predicted to be composed of two distinct protein modules. The C terminus seems to code for a catalytic domain belonging to a subfamily of adenylyl cyclase isozymes mostly found in Gram-positive bacteria. The expressed protein was shown to function as a homodimeric adenylyl cyclase (1 mol of cAMP⅐mg ؊1 ⅐min ؊1 ). In analogy to the structure of the mammalian adenylyl cyclase catalyst, six amino acids were targeted by point mutations and found to be essential for catalysis. The N-terminal region represents a novel protein domain, the occurrence of which is restricted to several adenylyl cyclases present in Grampositive bacteria. The purified full-length enzyme was 300-fold less active than the catalytic domain alone. (2), and in lower organisms, particularly in bacteria, class III CHDs seem to be linked with different protein domains that most likely impart peculiar regulatory features. However, so far only a few studies addressed bacterial class III ACs.In the completed genome of Mycobacterium tuberculosis, 15 open reading frames were identified that contain a CHD (2). The availability of this information enables us to study each mycobacterial AC isoform individually in vitro with the perspective to determine its contribution to the cAMP regulatory system during tuberculosis disease development in vivo. Two of the 15 AC open reading frames (Rv1625c and Rv2435c) belong to the mammalian-type ACs, and recent work concentrated on the membrane-bound mammalian-type AC present in Mycobacterium, Rv1625c (3, 4). Four predicted mycobacterial ACs (Rv1318c, Rv1319c, Rv1320c, and Rv3645) contain CHDs that are part of a subclass consisting of ACs from, among others, Anabaena, Stigmatella, Rhizobium, and Treponema (2). The remaining nine mycobacterial CHDs (Rv1264, Rv1647, Rv2212, Rv0386, Rv1358, Rv1359, Rv2488c, Rv0891c, and LipJ) are most similar to CHDs detected in other Gram-positive bacteria.Here we investigated the gene product of Rv1264. Earlier, AC genes of this subtype were identified by complementation cloning in Brevibacterium liquefaciens and Streptomyces (5, 6). Those genes code for modular proteins with the CHD located C-terminally. The dismal expression of these ACs in E. coli precluded detailed biochemical studies (5, 6). Thus, Rv1264 was used in an attempt to characterize this AC subtype. In addition, the biochemical characterization of the Rv1264 catalyst might constitute a starting point for future studies on the remaining eight related AC genes present in M. tuberculosis.We were able to express reasonable amounts of the Rv1264 AC catalytic domain in E. coli with high AC activity. The catalytic site is the result of homodimerization, and catalysis depends on the same amino acids previously identified as crucial in mammalian ACs. The holoenzyme was much less active than the catalytic domain alone, suggesting an autoinhibitory function of this unique N-terminal domain, which contains no similarity to any other known prot...

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A pyruvate‐stimulated adenylate cyclase has a sequence related to the fes/fps oncogenes and to eukaryotic cyclases

Cited by 19 publications

References 29 publications

Identification and Characterization ofglxR, a Gene Involved in Regulation of Glyoxylate Bypass inCorynebacterium glutamicum

Identification and Characterization ofglxR, a Gene Involved in Regulation of Glyoxylate Bypass inCorynebacterium glutamicum

The Conserved Asparagine and Arginine Are Essential for Catalysis of Mammalian Adenylyl Cyclase

Adenylyl Cyclase Rv1264 from Mycobacterium tuberculosis Has an Autoinhibitory N-terminal Domain

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