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Nucleic Acids Research, 1993, Vol. 21, No. 4 941-947
© 1993

MOLECULAR BIOLOGY

Rye nuclease I as a tool for structural studies of tRNAs with large variable arms

C.EI Adlouni, G. Keith*, G. Dirheimer, J. W. Szarkowski1 and A. Przykorska

Unité ‘Structure des MacromolÉcules Biologiques de Reconnaissance’, Unstitut de Biologie Moléculaire et Cellulaire du Centre National de la Recherche Scientiffique et UniversitéLouis Pasteur 15 Rue Descartes, 67084 Strasbourg, France 1Institute of Biochemistry and Biophysics, Polish Academy of Sciences Rakowiecka 36, 02-532 Warszawa, Poland

*To whom correspondence should be addressed

Received October 29, 1992. Revised January 15, 1993. Accepted January 15, 1993.

A single-strand-speclflc nuclease from rye germ (Rn nuclease I) was used for secondary and tertlary structure lnvestlgations of tRNAs with large variable arms (class II tRNAs). We have studied the structure In solutlon of two recently sequenced tRNALeu: yeast tRNALeu(ncm5UmAA) and bovine tRNALeu(XmAA) as well as yeast tRNALeu(UAG), tRNALeu(m5CAA) and tRNASer(IGA). The latter Is the only tRNA wlth a long variable arm for whlch the secondary and tertlary structure has already been studled by use of chemlcal probes and computer modelllng. The data obtalned in this work showed that the general model of class II tRNAs proposed by others for tRNAser can be extended to tRNAsLeu as well. However Interesting dlfferences in the structure of tRNAsLeU versus tRNASer(IGA) were also noticed. The maln difference was observed In the accessibility of the variable loops to nucleolytlc attack of Rn nuclease I : variable loops of all studied tRNALeu specles were cut by Rn nuclease I, while that of yeast tRNASer(IGA) was not. This could be due to differences In stablllty of the variable arms and the lengths of thelr loops which are 3 and 4 nucleotldes In tRNASer(IGA) and tRNALeu respectlvety.


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