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Nucleic Acids Research, 1982, Vol. 10, No. 4 1193-1201
© 1982

MOLECULAR BIOLOGY

Accessibility of U1 RNA to base pairing with a single-stranded DNA fragment mimicking the intron extremities at the splice junction

Eliane Lazar, Monique Jocob, Alain Krol and Christiane Branlant

Lab. Génétique Moléculaire des Eucaryotes du CNRS, U.184 de Biologie Moléculaire et de Génie Génétique, INSERM, Institut de Chimie Biologique 11, rue Humann, 67085 Strasbourg Cédex Institut de Biologie Moléculaire et Cellulaire du CNRS, Université Louis Pasteur 15, rue René Descartes, 67084 Strasbourg Cédex, France

Received December 22, 1981. Accepted January 18, 1982.

A DNA fragment containing a 16 nucleotide sequence mimicking the intron extremities of premessenger RNA aligned as proposed previously (1,2) in a model of splicing mechanism was prepared and used as a probe for accessibility of the 5' extremity of U1 RNA. Hybridization of U1 RNA to the probe under non denaturing conditions and digestion of the hybrid with RNase H revealed that the sequence of U1 RNA which is complementary to the extremities of introns is accessible to hybridization and to enzymes. Therefore, the configuration of isolated U1 RNA satisfies the criteria required for the alignment of introns and further enzymatic reactions of splicing.


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