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Nucleic Acids Research, 1982, Vol. 10, No. 23 7521-7529
© 1982

ENZYMOLOGY

Circularization of linear viroid RNA via 2'-phosphomonoester, 3', 5'-phosphodiester bonds by a novel type of RNA ligase from wheat germ and Chlamydomonas

Yo Kikuchi*, Kazimierz Tyc**, Witold Filipowicz§,**, Heinz L. Sänger# and Hans J. Gross

Institut für Biochemie Röntgenring 11, D-8700 Würzburg, FRG

Received October 15, 1982. Accepted November 9, 1982.

A novel type of RNA ligase activity in extracts of wheat germ or Chlamydomonas requires 2',3'-cyclic phosphate and 5'-phosphate ends for ligation to form a 2'-phosphomonoester, 3',5'-phosphodiester bond. Using 5'-32P-labeled linear PSTV, we demonstrate that RNase T1-nicked viroid predominantly forms FormulaU-bonds. Natural linear PSTV, however, forms main-FormulaA-bonds upon enzymatic circularization. We show that natural linear PSTV RNA has nicks between C181 and A182, or between C348 and A349, and that consequently C181 and C348 carry 2',3'-cyclophosphate termini.

*Mitsubishi-Kasei Institute of Life Sciences, Tokyo, Japan

**Institute of Biochemistry and Biophysics, Polish Academy of Science: Warsaw, Poland

§Roche Institute of Molecular Biology, Nutley, N.J., USA

#Max-Planck-Institut für Biochemie, D-8033 Martinsried, F.R.G.


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