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Nucleic Acids Research, 1978, Vol. 5, No. 8 2913-2926
© 1978

Articles

Studies on the single-stranded diseontinuities of the cauliflower mosaic virus genome

M. Volovitch*, G. Drugeon* and P. Yot

Section de Biologie Foundation Curie, 26 Rue d'Ulm, 75321 Paris Cedex 05, France *Institut de Recherche en Biologie Moleculaire du C.N.R.S., Université Paris VII 2 place Jussieu, 75221 PARIS CEDEX 05

Received March 31, 1978. The Cauliflower Mosaic Virus (CaMV) genome is a double-stranded DNA molecule of about 5 million daltons. Native DNA molecules appear heterogeneous when analysed by gel electrophoresis. We have examined the nature of this apparent heterogeneity. Besides, this genome is shown here to contain three single-stranded breaks, as revealed by different denaturation experimerits: heating at 75°C, treatment with NaOH or dimethyl sulfoxide (DMSO). Labelling with terminal transferase proves that the 3' ends at these interruptions all have free hydroxyl groups. Electron microscopy and alkaline gel electrophoresis indicate that these three discontinuities are shared by both strands, and that they are not randomly located. S1 nuclease is active on CaMV DNA and generates three fragments. The comparison between the sizes of these fragments and of the products of denaturation leads us to consider that S1 acts at the level of the interruptions. We have determined that two of them, distant by one third genome unit, are in the same strand; the other is in the opposite strand, distant by one sixth genome unit from the nearest other one. The combined use of restriction enzymes and S1 nuclease has enabled us to locate these three discontinuities on the restriction map of the CaMV genome that we have otherwise established.


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S. H. HOWELL, L. L. WALKER, and R. K. DUDLEY
Cloned Cauliflower Mosaic Virus DNA Infects Turnips (Brassica rapa)
Science, June 13, 1980; 208(4449): 1265 - 1267.
[Abstract] [PDF]


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