Cloning and characterization of the histidine biosynthetic gene cluster of Streptomyces coelicolor A3(2)

Limauro, Danila; Avitabile, A.; Cappellano, Carmela; Puglia, Anna Maria; Bruni, Carmelo B.

doi:10.1016/0378-1119(90)90436-u

Cited by 41 publications

(7 citation statements)

References 33 publications

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“…kluyveri HIS3 homolog. Bacterial HIS3 homologs from Azospirillum brasiliense (Fani et al, 1989), Escherichia coli (Chiariotti et al, 1986), Salmonella typhimurium (Carlomagno et al, 1988) and Streptomyces coelicolor (Limauro et al, 1990) are 4 5 4 7 % identical to the k-HIS3 O R F (696 nucleotides) at the nucleic acid level.…”

Section: Discussionmentioning

confidence: 99%

Molecular genetics in Saccharomyces kluyveri: The HIS3 homolog and its use as a selectable marker gene in S. kluyveri and Saccharomyces cerevisiae

Weinstock

Strathern

1993

Yeast

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We cloned the Saccharomyces kluyveri HIS3 homolog, k-HIS3, and made a partial deletion of the gene. The k-HIS3 gene complemented a HIS3 deletion in S. cerevisiae. The DNA sequences of the open reading frames (ORFs) of the HIS3 homologs are 70% identical at the DNA level and 83% identical at the deduced amino acid level. The ORF upstream of the k-HIS3 gene is related to the PET56 gene of S. cerevisiae found upstream of the HIS3 gene of S. cerevisiae. The ORF downstream from the k-HIS3 gene is not related to the DED1 gene found downstream of the HIS3 gene in S. cerevisiae.

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Section: Discussionmentioning

confidence: 99%

Molecular genetics in Saccharomyces kluyveri: The HIS3 homolog and its use as a selectable marker gene in S. kluyveri and Saccharomyces cerevisiae

Weinstock

Strathern

1993

Yeast

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“…Figure 5 shows the complete progress curve for conversion of IGP into IAP, both in the presence and in the absence of 0.1 M ,6-mercaptoethanol. Comparing the amino acid sequences of IGPD derived from the DNA sequences of S. cerevisiae (Struhl, 1985), Azospirillum brasilense (Fani et al, 1989), S. typhimurium (Carlomagno et al, 1988), Streptomyces coelicum (Limauro et al, 1990), Trichoderma harzianum (Goldman et al, 1992) and E. coli (Chiariotti et al, 1986), there is no conserved cysteine residue. It seems, unlikely, therefore that a cysteine residue has a critical role in any evolutionarily conserved mechanism.…”

Section: Properties Of Mn-igpdmentioning

confidence: 99%

Purification and characterization of the imidazoleglycerol-phosphate dehydratase of Saccharomyces cerevisiae from recombinant Escherichia coli

Hawkes

Thomas

Edwards

et al. 1995

Biochemical Journal

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The HIS3+ gene of Saccharomyces cerevisiae was overexpressed in Escherichia coli and the recombinant imidazoleglycerol-phosphate dehydratase (IGPD) purified to homogeneity. Laser-desorption and electrospray m.s. indicated a molecular ion within 2 units of that expected (23833.3) on the basis of the protein sequence, with about half of the polypeptide lacking the N-terminal formylmethionine residue. IGPD initially purified as an apoprotein was catalytically inactive and mainly a trimer of M(r) 70,000. Addition of Mn2+ (but not Mg2+) caused this to assemble to an active (40 units/mg) enzyme (Mn-IGPD) comprising of 24 subunits (M(r) 573,000) and containing 1.35 +/- 0.1 Mn atoms/polypeptide subunit. An enzyme with an identical activity and metal content was also obtained when the fermenter growth medium of recombinant Escherichia coli was supplemented with MnCl2, and IGPD was purified through as Mn-IGPD rather than as the apoenzyme and assembled in vitro. Inhibition by EDTA indicated that the intrinsic Mn2+ was essential for activity. The retention of activity over time after dilution to very low concentrations of enzyme (< 20 nM) indicated that the metal remained in tight association with the protein. A novel continuous assay method was developed to facilitate the kinetic characterization of Mn-IGPD. At pH 7.0, the Km for IGP was 0.10 +/- 0.02 mM and the Ki value for inhibition by 1,2,4-triazole, 0.12 +/- 0.02 mM. In contrast with other reports, thiols had no influence on catalytic activity. The activity of Mn-IGPD varied with enzyme concentration in such a way as to suggest that it dissociates to a less active form at very low concentrations. Significant inhibition by the product, imidazole acetol phosphate, was inferred from the shape of the progress curve. Titration with, the potent competitive inhibitor, 2-hydroxy-3-(1,2,4-triazol-1-yl)propyl phosphonate indicated that Mn-IGPD contained 0.9 +/- 0.1 catalytic sites/protomer. The activity nearly doubled in the presence of high concentrations of Mn2+; the apparent Ks for stimulation was 20 microM. The basis of this effect was obscure, since there was no corresponding increase in the titre of active sites. Neither was there a discernable shift in the values of Km or Ki (above), although exogenous Mn2+ did reduce the optimum pH for kcat, from 7.2 to 6.8. On the basis of a single site/subunit, the maximum rate of catalytic turnover at 30 degrees C was 32 s-1.

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“…DNA sequence analysis showed that AG29 harbored a hybrid plasmid insert (base pairs 13638-18321 of cosmid SC4G6; Fig. 1) within the histidine biosynthesis operon (44). Consistent with this, the mutant was able to grow on minimal medium that had been supplemented with histidine (not shown).…”

Section: An Insertion Mutation Blocking Spore Formation At An Early Smentioning

confidence: 58%

Genomewide insertional mutagenesis in Streptomyces coelicolor reveals additional genes involved in morphological differentiation

Gehring

Nodwell

Beverley

et al. 2000

Proc. Natl. Acad. Sci. U.S.A.

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The filamentous soil bacterium Streptomyces coelicolor undergoes a complex cycle of morphological differentiation involving the formation of an aerial mycelium and the production of pigmented antibiotics. We have developed a procedure for generating insertional mutants of S. coelicolor based on in vitro transposition of a plasmid library of cloned S. coelicolor DNAs. The insertionally mutated library was introduced into S. coelicolor, and transposon insertions were recovered at widely scattered locations around the chromosome. Many of the insertions revealed previously uncharacterized genes, and several caused novel mutant phenotypes, such as altered pigment production, enhanced antibiotic sensitivity, delayed or impaired formation of aerial hyphae, and a block in spore formation. The sporulation mutant harbored an insertion in one of three adjacent genes that are apparently unique to Streptomyces but are each represented by at least 20 paralogs at dispersed locations in the chromosome. Individual members of the three families often are found grouped together in a characteristic arrangement, suggesting that they have a common function.

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Cloning and characterization of the histidine biosynthetic gene cluster of Streptomyces coelicolor A3(2)

Cited by 41 publications

References 33 publications

Molecular genetics in Saccharomyces kluyveri: The HIS3 homolog and its use as a selectable marker gene in S. kluyveri and Saccharomyces cerevisiae

Molecular genetics in Saccharomyces kluyveri: The HIS3 homolog and its use as a selectable marker gene in S. kluyveri and Saccharomyces cerevisiae

Purification and characterization of the imidazoleglycerol-phosphate dehydratase of Saccharomyces cerevisiae from recombinant Escherichia coli

Genomewide insertional mutagenesis in Streptomyces coelicolor reveals additional genes involved in morphological differentiation

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