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Nucleic Acids Research, 1982, Vol. 10, No. 4 1215-1229
© 1982

MOLECULAR BIOLOGY

The orderly splicing of the first three leaders of the adenovirus-2 major late transcript

P. Keohavong, R. Gattoni, J.M. LeMoullec*, M. Jacob and J. Stévenin

Unité 184 de Biologie Moléculaire et de Génie Génétique de l'INSERM, et Laboratoire de Génétique Moléculaire des Eucaryotes du CNRS, Institut de Chimie Biologique, Faculté de Médecine 11, rue Humann, 67085 Strasbourg Cédex, France

Received December 7, 1981. Revised January 27, 1982. Accepted January 27, 1982.

A strategy based on the hybridization of labeled nucleoplasmic RNA to a short cloned cDNA probe was devised to study the ligation of the three first leader sequences (Le1, Le2, Le3) of the major late adenovirus-2 transcript. The hybridized RNA was subsequently fractionated by electrophoresis and identified with the aid of restriction fragments of the DNA probe. The ligations were shown to occur stepwise and in an orderly fashion. Le1 and Le2 were first ligated without detectable lag time. The tripartite leader was formed after a lag time of 10–15 min probably due, for a large part, to the stepwise excision of the intervening sequence between Le2 and Le3. The possible processing intermediate Le2-Le3 was not detected.

*Unité 163 de Recombinaisons et Expression Génétique de l'INSERM, Institut Pasteur, 25, rue du Dr. Roux, 75015 Paris, France


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