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J. Biol. Chem., Vol. 275, Issue 47, 37278-37284, November 24, 2000
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From the Friedrich-Miescher-Institute, P.O. Box 2543, CH-4002 Basel, Switzerland
The pathways of scanning ribosome migration
controlled by the cauliflower mosaic virus 35 S RNA leader were
investigated in vitro and in vivo. This long
(600 nucleotides) leader contains several short open reading frames
(sORFs) and folds into an extended hairpin structure with three main
stable stem sections. Translation initiation downstream of the leader
is cap-dependent and occurs via ribosomal shunt under the
control of two cis elements, a short open reading frame A
(sORF A) followed by stem section 1. Here we show that a second similar
configuration comprising sORF B followed by stem section 2 also allows
shunting. The efficiency of the secondary shunt was greatly increased
when stem section 1 was destabilized. In addition, we present evidence
that a significant fraction of reinitiation-competent ribosomes that
escape both shunt events migrate linearly via the structured central
region but are intercepted by internal AUG start codons. Thus,
expression downstream of the 35 S RNA leader is largely controlled by
its multiple sORFs.
Continuous and Discontinuous Ribosome Scanning on the Cauliflower
Mosaic Virus 35 S RNA Leader Is Controlled by Short Open Reading
Frames*
,
*
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.
Supported by a Federation of European Biochemical Societies fellowship.
§
Present address: Center for Human Genetics, Katholieke
Universiteit Leuven; Herestraat 49, 3000 Leuven, Belgium.
¶
To whom correspondence should be addressed. Tel.: 4161-697 66 84; Fax: 4161-697 39 76; E-mail: Hohn@fmi.ch.
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