Characterization of the glyoxysomal isocitrate lyase genes of Aspergillus nidulans (acuD) and Neurospora crassa (acu-3)

Gainey, L. D. S.; Connerton, Ian F.; Lewis, Evelyn L.; Turner, Geoffrey; Ballance, D. J.

doi:10.1007/bf00318653

Cited by 49 publications

(31 citation statements)

References 31 publications

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“…The comparison of the deduced ICL protein sequence from E. coli with those from eucaryotic organisms revealed a high level of similarity between the E. coli and the eucaryotic enzymes. However, the E. coli enzyme turned out to be about 16 kDa smaller than those from eucaryotes (33). This result is in agreement with the findings that by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis, most ICLs from eucaryotes are significantly larger than their procaryotic counterparts (for a review, see reference 46).…”

supporting

confidence: 87%

Characterization of the isocitrate lyase gene from Corynebacterium glutamicum and biochemical analysis of the enzyme

1994

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Isocitrate lyase is a key enzyme in the glyoxylate cycle and is essential as an anaplerotic enzyme for growth on acetate as a carbon source. It is assumed to be of major importance in carbon flux control in the amino acid-producing organism Corynebacterium ghltaml'cum. In crude extracts of C. glutamicum, the specific activities of isocitrate lyase were found to be 0.01 U/mg of protein after growth on glucose and 2.8 U/mg of protein after growth on acetate, indicating tight regulation. The isocitrate lyase gene, aceA, was isolated, subcloned, and characterized. The predicted gene product of aceA consists of 432 amino acids (M1, 47,228) (27, 28; for a review, see reference 46). Particularly in Escherichia coli, regulation of carbon flux between the two cycles has been subject to intensive investigation. It was found that in E. coli, ICL is formed only when acetate is the sole carbon source of the medium (24). ICL shows low affinity towards isocitrate, and ICD exhibits high affinity for its substrate. To allow significant amounts of carbon to enter the glyoxylate cycle, ICD in E. coli is substantially inactivated by reversible phosphorylation during growth on acetate (27,28,49). Both phosphorylation and dephosphorylation is catalyzed by one enzyme, ICD kinase/phosphatase (28). When acetate is the sole carbon source, the kinase activity of this enzyme leads to phosphorylation and thus to inactivation of ICD. When glucose is added, ICD is dephosphorylated, resulting in the restoration of ICD activity. Besides this regulation of ICD, the E. coli ICL is activated by phosphorylation (37) and is inhibited by a variety of metabolites, e.g., phosphoenolpyruvate (PEP), 3-phosphoglycerate, and succinate (31, 38), thereby allowing fine control of the carbon flux.Since ICL has a key position in the central metabolism, increased attention has been focused on the genetics of ICL in

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confidence: 87%

Characterization of the isocitrate lyase gene from Corynebacterium glutamicum and biochemical analysis of the enzyme

1994

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“…The carboxy-terminal sequence of isocitrate lyase from P. pastoris does not coincide with the eukaryotic signal S(A,C)-K(H,R)-L for import of the protein into the peroxisome. However, peroxisomal Icl from Aspergillus nidulans and Neurospora crassa lack the PTS1-like carboxy-terminal amino acids S(A,C)-K(H,R)-L (Gainey et al, 1992). Therefore, conclusive evidence for the localization of the enzyme in P. pastoris can only be obtained from further analysis.…”

Section: Resultsmentioning

confidence: 99%

The ICL1 gene of Pichia pastoris, transcriptional regulation and use of its promoter

Menéndez¹,

Valdés²,

Cabrera³

2003

Yeast

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We cloned and characterized a gene encoding isocitrate lyase from the methylotrophic yeast Pichia pastoris. This gene was isolated from a P. pastoris genomic library using a homologous PCR hybridization probe, amplified with two sets of degenerate primers designed from conserved regions in yeast isocitrate lyases. The cloned gene was sequenced and consists of an open reading frame of 1563 bp encoding a protein of 551 amino acids. The molecular mass of the protein is calculated to be 60.6 kDa with high sequence similarity to isocitrate lyase from other organisms. There is a 64% identity between amino acid sequences of P. pastoris Icl and Saccharomyces cerevisiae Icl. Northern blot analyses showed that, as in S. cerevisiae, the steady-state ICL1 mRNA levels depend on the carbon source used for cell growth. Expression in P. pastoris of the dextranase gene (dexA) from Penicillium minioluteum under control of the ICL1 promoter proved that P ICL1 is a good alternative for the expression of heterologous proteins in this methylotrophic yeast. The sequence presented here has been deposited in the EMBL data library under Accession No. AJ272040.

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“…By analogy with acu-D, the ICLs of A. nidulans (Gainey et al, 1992), D111, D165, D167 and E194 are suggested to be a cluster for the Mg 2+ -binding site. The predicted amino acid sequence was compared with those of the ICLs of Phanerochaete chrysosporium, C. cinereus (Chaure et al, 1997), A. nidulans (Gainey et al, 1992) and Saccharomyces cerevisiae (Schöler & Schüller, 1993), which revealed 83, 79, 63 and 54 % identity, respectively (Fig. 1).…”

Section: Characterization Of Fpicl1mentioning

confidence: 99%

Subcellular localization of glyoxylate cycle key enzymes involved in oxalate biosynthesis of wood-destroying basidiomycete Fomitopsis palustris grown on glucose

et al. 2006

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This study investigated the subcellular localization of key enzymes of the glyoxylate cycle, i.e. isocitrate lyase (ICL; EC 4.1.3.1) and malate synthase (EC 2.3.3.9), that function constitutively in coordination with oxalate biosynthesis of glucose-grown Fomitopsis palustris. The ICL purified previously from F. palustris is termed FPICL1. Subcellular fractionation analysis of the cell homogenate by the sucrose density-gradient method showed that both key enzymes were present in peroxisomes, whereas acetyl-CoA synthase (EC 6.2.1.1) and oxalate-producing oxaloacetate acetylhydrolase (EC 3.7.1.1) were cytosolic. The peroxisomal localization of FPICL1 was further confirmed by electron microscopic and immunocytochemical analysis with anti-FPICL1 antibody. In addition, the peroxisomal target signal, composed of SKL at the C terminus of the cDNA encoding FPICL1, was found, which also suggests that FPICL1 is peroxisomal. Accordingly, it is postulated that transportation of succinate from peroxisomes to mitochondria, and vice versa, for the transportation of isocitrate or citrate, occurs in glucose-grown F. palustris for the constitutive metabolic coordination of the TCA and glyoxylate cycles with oxalate biosynthesis.

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Characterization of the glyoxysomal isocitrate lyase genes of Aspergillus nidulans (acuD) and Neurospora crassa (acu-3)

Cited by 49 publications

References 31 publications

Characterization of the isocitrate lyase gene from Corynebacterium glutamicum and biochemical analysis of the enzyme

Characterization of the isocitrate lyase gene from Corynebacterium glutamicum and biochemical analysis of the enzyme

The ICL1 gene of Pichia pastoris, transcriptional regulation and use of its promoter

Subcellular localization of glyoxylate cycle key enzymes involved in oxalate biosynthesis of wood-destroying basidiomycete Fomitopsis palustris grown on glucose

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