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Plant Physiol, May 2001, Vol. 126, pp. 167-175
The Internal Propeptide of the Ricin Precursor Carries a
Sequence-Specific Determinant for Vacuolar Sorting1
Lorenzo
Frigerio,*
Nicholas A.
Jolliffe,
Alessandra
Di Cola,
Doramys Hernández
Felipe,
Nadine
Paris,
Jean-Marc
Neuhaus,
J.
Michael
Lord,
Aldo
Ceriotti, and
Lynne M.
Roberts
Department of Biological Sciences, University of Warwick, Coventry
CV4 7AL, United Kingdom (L.F., N.A.J., A.D.C., J.M.L., L.M.R.);
Laboratorie de Biochimie, Université de Neuchâtel, rue
Émile-Argand 9, CH-2007 Neuchâtel 7, Switzerland (D.H.F.,
N.P., J.-M.N.); and Istituto Biosintesi Vegetali, Consiglio Nazionale
delle Ricerche, via Bassini 15, 20133 Milano, Italy (A.C.)
Ricin is a heterodimeric toxin that accumulates in the storage
vacuoles of castor bean (Ricinus communis) endosperm.
Proricin is synthesized as a single polypeptide precursor comprising
the catalytic A chain and the Gal-binding B chain joined by a 12-amino acid linker propeptide. Upon arrival in the vacuole, the linker is
removed. Here, we replicate these events in transfected tobacco (Nicotiana tabacum) leaf protoplasts. We show that the
internal linker propeptide is responsible for vacuolar sorting and is
sufficient to redirect the ricin heterodimer to the vacuole when fused
to the A or the B chain. This internal peptide can also target two different secretory protein reporters to the vacuole. Moreover, mutation of the isoleucine residue within an NPIR-like motif of the propeptide affects vacuolar sorting in proricin and in the reconstituted A-B heterodimer. This is the first reported example of a
sequence-specific vacuolar sorting signal located within an internal propeptide.
1
This work was supported in part by the European
Union (grant no. CHRX-CT94-0590), by the Biotechnology and Biological
Science Research Council (grant no. 88/C08612), by the British
Council/Ministero dell'Universita'e della Ricerca Scientifica e
Tecnologica (grant no. ROM/889/99/10), and by the Swiss National
Science Foundation (grant no. 31-46926.96).
*
Corresponding author; e-mail l.frigerio{at}warwick.ac.uk; fax
44-2476-523701.
© 2001 American Society of Plant Physiologists
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