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Nucleic Acids Research, 1993, Vol. 21, No. 6 1361-1367
© 1993

MOLECULAR BIOLOGY

Role of the 5' leader sequence of alfalfa mosaic virus RNA 3 in replication and translation of the viral RNA

Edwin A.G. Van der Vossen, Lyda Neeleman and John F. Bol*

Institute of Molecular Plant Sciences, Gorlaeus Laboratories, Leiden University PO Box 9502, 2300 RA Leiden, The Netherlands

*To whom correspondence should be addressed

Received January 8, 1993. Revised February 16, 1993. Accepted February 16, 1993.

RNA 3 of alfalfa mosaic virus (AIMV) encodes the movement protein P3 and the viral coat protein which is translated from the subgenomic RNA 4. The 5'-leader sequences of RNA 3 of AIMV strains S, A, and Y differ in length from 314 to 392 nucleotldes and contain a variable number of internal control regions of type 2 (ICR2 motifs) each located In a 27 nt repeat. Infectious cDNA clones were used to exchange the leader sequences of the three strains. This revealed that the leader sequence controls the specific ratio In which RNAs 3 and 4 are synthesized for each strain. In addition, It specifies strain specific differences in the kinetics of P3 accumulation In plants. Subsequent deletion analysis revealed that a 5'-sequence of 112 nt containing one ICR2 motif was sufficient for a 10 to 20% level of RNA 3 accumulation in protoplasts and a delayed accumulation In plants. An additional leader sequence of maximally 114 nt, containing two ICR2 motifs, was required to permit wlldtype levels of RNA 3 accumulation. The effect of deletions In the leader sequence on P3 synthesis In vitro and In vivo was Investigated.


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