Changes in macromolecular synthesis in Xanthomonas oryzae infected with bacteriophage XP-12

Ehrlich, Melanie; Lin, Fu-Pang; Ehrlich, Kenneth C.; Brown, Susan L.; Mayo, John A.

doi:10.1128/jvi.23.3.517-523.1977

Cited by 8 publications

(7 citation statements)

References 33 publications

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“…Cm5dCyd (1.00 A287 -0.32 A267) x 10-4 CdThd (1.34 A267 -0.69 A287) x Because the bisulfite adducts of pyrimidines have negligible absorption at greater than 240 nm (12), the amount of bisulfite adduct remaining can be determined from the difference between the original concentration of the m5dCyd and the sum of the concentrations of m5dCyd and dThd after the reaction. DNA Preparation and DNA-bisulfite reaction: For preparing X. oryzae DNA exclusively radiolabeled in its Cyt residues, a thymine auxotroph of X. oryzae was grown in XMS medium (19) supplemented with dThd and [2-14C]uracil (21). To label bacteriophage DNA to equal specific activity in Thy and m5Cyt residues, the phage were propagated on X. oryzae in the presence of [2-14C]uracil as previously described (21).…”

Section: Methodsmentioning

confidence: 99%

“…DNA Preparation and DNA-bisulfite reaction: For preparing X. oryzae DNA exclusively radiolabeled in its Cyt residues, a thymine auxotroph of X. oryzae was grown in XMS medium (19) supplemented with dThd and [2-14C]uracil (21). To label bacteriophage DNA to equal specific activity in Thy and m5Cyt residues, the phage were propagated on X. oryzae in the presence of [2-14C]uracil as previously described (21). Unlabeled DNA was isolated by standard methods (22).…”

Section: Methodsmentioning

confidence: 99%

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Comparison of bisulfite modification of 5-methyldeoxycytidine and deoxycytidine residues

1980

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Sodium bisulfite is a mutagen which can specifically deaminate more than 96% of the cytosine residues in single-stranded DNA via formation of a 5,6-dihydrocytosine-6-sulfonate intermediate. Under the same reaction conditions, only 2-3% of the 5-methylcytosine (m5Cyt) residues in single-stranded XP-12 DNA, which has 34 mole% m5Cyt, was converted to thymine (Thy) residues. In contrast, at the deoxynucleoside and free base levels, the same treatment with bisulfite and then alkali converted 51% and > 95%, respectively, of the m5Cyt to the corresponding Thy derivatives. However, the rate of reaction of m5Cyt and its deoxyribonucleoside was much slower than that of the analogous quantitative conversion of cytosine or deoxycytidine to uracil or deoxyuridine, respectively. The much lower reactivity of m5Cyt and its derivatives compared to that of the unmethylated analogs is primarily due to a decrease in the rate of formation of the sulfonate adduct.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Comparison of bisulfite modification of 5-methyldeoxycytidine and deoxycytidine residues

1980

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“…Phage-infected bacteria were harvested 45 min postinfection in XPS medium (3) according to previous methods (6). X. oryzae [14C]cytosine DNA was prepared from the Thy auxotroph X. oryzae thyH (4) grown in medium containing 1 ,Ci of [2-14C]uracil per ml (30 mCi/mmol, ICN Pharmaceuticals, Irvine, Calif.) and 5 ,ug of unlabeled dThd per ml. XP-12 [14C]pyrimidine DNA, that is, DNA labeled to equal specific activity in both pyrimidine residues, was obtained from XP-12-infected X. oryzae propagated in the same medium containing [2-14C]uracil without dThd.…”

Section: Methodsmentioning

confidence: 99%

“…When a Thy auxotrophic host is transferred to a medium deficient in dThd and then infected with XP-12, a normal yield of phage is obtained. Nonetheless, in this medium the uninfected host is committed to thymineless death (4). When XP-12 phage is propagated on this Thy auxotroph in medium containing [2-14C]uracil and 50 ,ug of dThd per ml, the resulting phage DNA contains m5Cyt and Thy residues labeled to equal specific activity.…”

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confidence: 99%

5-methyl-dCTP deaminase induced by bacteriophage XP-12

Wang¹,

Ehrlich²

1982

J Virol

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Bacteriophage XP-12, whose DNA contains 34 mol% 5-methylcytosine, induces the synthesis of a unique enzyme, 5-methyl-dCTP deaminase. The substrate for this enzyme, 5-methyl-dCTP, is produced by reactions catalyzed in part by other phage-induced enzymes, and the product of the reaction is dTTP. The deaminase therefore provides a novel pathway for biosynthesis of thymine residues for phage XP-12 DNA. Evidencd is presented that this pathway is used for dTTP synthesis in XP-12-infected Xanthomonas oryzae.

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“…oryzae, Xp12 is the only known source of DNA in which cytosine is completely replaced by S-methylcytosine [23]. This unusual property has made Xp12 a unique model for probing the effects of naturally occurring methylation of DNA cytosine residues [15,24]. A significant change in the pattern of protein phosphorylation has been discovered in X. oryzae pv.…”

Section: Introductionmentioning

confidence: 99%

Rifampicin: an inhibitor of Xpl2-specific protein phosphorylation in Xanthomonas oryzae pv. oryzae

Cheng

Yang

et al. 1996

FEMS Microbiology Letters

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Phosphorylation of the three Xp12-specific phosphoproteins was drastically reduced by rifampicin, an antibiotic that specifically inhibits the host-cell RNA polymerase. However, this inhibitory effect could not be found in spontaneous mutants of Xanthomonas oryzae pv. oryzae whose RNA polymerase are resistant to the drug. The inhibitory effect of rifampicin treatment also resulted suppression of the Xp12 multiplication cycle. This implies the physiological significance of this effect and supports our previous prediction that phosphorylation plays an important role in the life cycle of Xp12. The acid- and alkali-labile character of the Xp12-specific phosphoproteins and the chemical stability of the phosphoryl linkages show that the corresponding protein kinase catalyzes the formation of an acyl phosphorylation. Subsequent fractionation of cell lysate revealed that the phosphoproteins were located in the periplasm. Actinomycin D, which affects transcription through DNA condensation rather than its binding to RNA polymerase, was not able to cause the inhibition effect. On the other hand, cerulenin was found to reduce the acyl phosphorylation which hints at a possible role of cell membrane in the phosphorylation. Here we present the evidence for the functional involvement of the rifampicin treatment on protein phosphorylation. A possible mechanism of rifampicin on the alternation of acyl phosphorylation is proposed.

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Changes in macromolecular synthesis in Xanthomonas oryzae infected with bacteriophage XP-12

Cited by 8 publications

References 33 publications

Comparison of bisulfite modification of 5-methyldeoxycytidine and deoxycytidine residues

Comparison of bisulfite modification of 5-methyldeoxycytidine and deoxycytidine residues

5-methyl-dCTP deaminase induced by bacteriophage XP-12

Rifampicin: an inhibitor of Xpl2-specific protein phosphorylation in Xanthomonas oryzae pv. oryzae

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