Nucleic Acids Research, 1988, Vol. 16, No. 22 10717-10732
© 1988
MOLECULAR BIOLOGY |
Exploration of the L18 binding site on 5S RNA by deletion mutagenesis
1Departments of Molecular Biophysics and Biochemistry, Yale University New Haven, CT 06511, USA 2Department of Chemistry, Yale University New Haven, CT 06511, USA
*To whom correspondence should be addressed
Received June 17, 1988. Revised October 21, 1988. Accepted October 21, 1988.
Several deletion variants of E. coli 5S RNA have been constructed and produced either in vivo or in vitro using T7 RNA Polymerase. Their structures and ribosomal protein L18 binding properties have been examined. All of them are similar to wild-type 5S RNA in their helix II-III regions, where L18 binds [Huber, P.W. and Wool, LG. (1984) Proc. Natl. Acad. Sci. (USA) 81, 322–326; Douthwaite, S., Christensen, A., and Garrett, R.A. (1982) Biochemistry 21, 2313–2320.], by NMR criteria. However, none of the molecules examined that lack the helix IV-helix V stem bind L18 efficiendy, even though that portion of 5S RNA is outside tbe L18 footprint. Tbe L18 binding site is clearly more than a simple hairpin loop.
+Present address: Department of Biochemistry and Molecular Biology, Harvard University, Cambridge, MA 02138, USA