Details

Autor(en) / Beteiligte

• Biswal, Nilambar
• Murray, Byron K.
• Benyesh-Melnick, Matilda

Titel

Ribonucleotides in newly synthesized DNA of herpes simplex virus

Ist Teil von

• Virology (New York, N.Y.), 1974-09, Vol.61 (1), p.87-99

Ort / Verlag

United States: Elsevier Inc

Erscheinungsjahr

1974

Quelle

Elsevier Journal Backfiles on ScienceDirect (DFG Nationallizenzen)

Beschreibungen/Notizen

• Newly synthesized DNA of herpes simplex virus type 1 (HSV-1), obtained from primary rabbit kidney cells pulse-labeled with [ 3H]thymidine or [ 3H]uridine at 6 hr postinfection, was purified by two cycles of centrifugation in CsCl density gradients. These intracellular viral DNA preparations hybridized specifically with homologous HSV-1 DNA but not with host cell DNA or E. coli DNA. Upon denaturation by alkali, the [ 3H]thymidine-labeled HSV-1 DNA cleaved to smaller pieces. The alkali-labile material in the viral DNA was identified as ribonucleotides on the basis of the following observations: (1) When labeled with [ 3H]uridine for short periods, the labeled viral “DNA” was susceptible to RNase and NaOH, and all the radioactivity was confined to the nucleoside [ 3H]uridine; however, upon longer labeling periods (up to 20 hr), the [ 3H]uridine-labeled viral “DNA” became more susceptible to DNase, as most but not all of the [ 3H]uridine was converted to deoxyribonucleosides. (2) Denaturation of [ 3H]uridine-labeled double-stranded HSV-1 “DNA” ( ϱ Cs 2SO 4 = 1.45 g/cm 3) by heat shifted the buoyant density to single-stranded DNA region ( ϱ Cs 2SO 4 = 1.48−1.50 g/cm 3) but not to single-stranded RNA region; however, treatment with hot NaOH considerably reduced the radioactivity of this “DNA.” Treatment with DNase, but not with pronase, shifted the buoyant density to the heavier RNA region of the gradient. Heat-denatured DNA but not the native DNA was susceptible to single-strand specific nuclease S1.