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J Biol Chem, Vol. 275, Issue 2, 1269-1274, January 14, 2000
From the The antisense Inc RNA encoded by the IncI
Structural Analysis of Late Intermediate Complex Formed
between Plasmid ColIb-P9 Inc RNA and Its Target RNA
HOW DOES A SINGLE ANTISENSE RNA REPRESS TRANSLATION OF
TWO GENES AT DIFFERENT RATES?*
§ and Department of Biophysics and
Biochemistry, Graduate School of Science, University of Tokyo,
Hongo, Tokyo 113-0033, Japan and the ¶ Department of Applied
Physics and Chemistry, University of Electro-Communications,
Chofu-shi, Tokyo 182-8585, Japan
ColIb-P9 plasmid replicon controls the translation of repZ
encoding the replication initiator and its leader peptide
repY at different rates with different mechanisms. The
initial loop-loop base pairing between Inc RNA and the target in the
repZ mRNA leader inhibits formation of a pseudoknot
required for repZ translation. A subsequent base pairing at
the 5' leader of Inc RNA blocks repY translation. To delineate the molecular basis for the differential control, we analyzed
the intermediate complexes formed between RepZ mRNA and Inc
RNA54, a 5'-truncated Inc RNA derivative. We found that the initial base pairing at the loops transforms into a more stable intermediate complex by its propagation in both directions. The resulting extensive base pairing indicates that the inhibition of the
pseudoknot formation is established at this stage. Furthermore, the
region of extensive base pairing includes bases different in related
plasmids showing different incompatibility. Thus, the observed
extensive base pairing is important for determining the incompatibility
of the low-copy-number plasmids. We discuss the evolution of
replication control systems found in IncI, IncB, and IncFII group plasmids.
*
This research was supported by a grant-in-aid from the
Ministry of Education, Science, Sports and Culture (Monbu-sho) of
Japan.The costs of publication of this
article were defrayed in part by the
payment of page charges. The article
must therefore be hereby marked
"advertisement" in
accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
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