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Molecular and Cellular Biology, September 1998, p. 5021-5031, Vol. 18, No. 9
0270-7306/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.
Sendai Virus Y Proteins Are Initiated by a
Ribosomal Shunt
Patrizia
Latorre,
Daniel
Kolakofsky, and
Joseph
Curran*
Department of Genetics and Microbiology,
University of Geneva Medical School (CMU), CH1211 Geneva,
Switzerland
Received 9 February 1998/Returned for modification 24 March
1998/Accepted 25 May 1998
The Sendai virus P/C mRNA expresses eight primary translation
products by using a combination of ribosomal choice and
cotranscriptional mRNA editing. The longest open reading frame (ORF) of
the mRNA starts at AUG104 (the second initiation site) and
encodes the 568-amino-acid P protein, an essential subunit of the viral
polymerase. The first (ACG81), third (ATG114),
fourth (ATG183), and fifth (ATG201) initiation
sites are used to express a C-terminal nested set of polypeptides
(collectively named the C proteins) in the +1 ORF
relative to P, namely, C', C, Y1, and Y2, respectively. Leaky scanning accounts for translational initiation at the first three start
sites (a non-ATG followed by ATGs in progressively stronger contexts).
Consistent with this, changing ACG81/C' to ATG
(GCCATG81G) abrogates expression from the
downstream ATG104/P and ATG114/C initiation
codons. However, expression of the Y1 and Y2 proteins remains normal in
this background. We now have evidence that initiation from
ATG183/Y1 and ATG201/Y2 takes place via a
ribosomal shunt or discontinuous scanning. Scanning complexes appear to
assemble at the 5' cap and then scan ca. 50 nucleotides (nt) of the 5'
untranslated region before being translocated to an acceptor site at or
close to the Y initiation codons. No specific donor site sequences are
required, and translation of the Y proteins continues even when
their start codons are changed to ACG. Curiously,
ATG codons (in good contexts) in the P ORF, placed either 16 nt
upstream of Y1, 29 nt downstream of Y2, or between the Y1 and
Y2 codons, are not expressed even in the
ACGY1/ACGY2 background. This indicates that
ATG183/Y1 and ATG201/Y2 are privileged start
sites within the acceptor site. Our observations suggest that the shunt
delivers the scanning complex directly to the Y start codons.
*
Corresponding author. Mailing address: Department of
Genetics and Microbiology, University of Geneva Medical School (CMU), 9 Ave. Champel, CH1211 Geneva, Switzerland. Phone: 41-22-702-57.27. Fax:
41-22-346-72.37. E-mail: curran{at}cmu.unige.ch.
Molecular and Cellular Biology, September 1998, p. 5021-5031, Vol. 18, No. 9
0270-7306/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.
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