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Nucleic Acids Research, 1979, Vol. 6, No. 5 1747-1760
© 1979

Articles

The regulatory region of MS2 phage RNA repliwse cistron. III. Characterization of fragments resulting from S1 nuclese digestion

I. Jansone, V. Berzin, V. Gribanov and E.J. Gren

Laboratory of Nucleic Acid Chemistry, Institute of Organic Synthesis, Latvian Academy of Sciences Riga 226006, USSR

Received March 15, 1979. The functionally active fragments MS2 R(-53<-6) and MS2 R(-53<-3) comprising the regulatory region for the replicase cistron have been isolated from MS2 RNA-coat protein complex following TI RNase digestion. In order to obtain shorter fragments, active in coat protein binding and initiation of translation, MS2 R(-53<-6) was cleaved with S1 nuclease. The results indicate that Sl nuclease attacks the most susceptible loop regions of the two hairpin helices of MS2 R(-53<-6). Amon the three fragments which have been isolated, only W 2 R(-35/33<-6) containing the intact hairpin (b) region with initiation codon AUG is active in the coat protein binding. Functional activity exerted by another polynucleotide MS2 R(-17<-6) supports the assumption that specific binding with the coat protein is determined by the hairpin (b) region prior to the replicase cistron.


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