The N-terminal ricin propeptide influences the fate of ricin A-chain in tobacco protoplasts.
The plant toxin ricin is synthesized in castor bean seeds as an endoplasmic reticulum (ER)-targeted precursor. Removal of the signal peptide generates proricin in which the mature A- and B-chains are joined by an intervening propeptide and a 9-residue propeptide persists at the N terminus. The two propeptides are ultimately removed in protein storage vacuoles, where ricin accumulates. Here we have demonstrated that the N-terminal propeptide of proricin acts as a nonspecific spacer to ensure efficient ER import and glycosylation. Indeed, when absent from the N terminus of ricin A-chain, the non-imported material remained tethered to the cytosolic face of the ER membrane, presumably by the signal peptide. This species appeared toxic to ribosomes. The propeptide does not, however, influence catalytic activity per se or the vacuolar targeting of proricin or the rate of retrotranslocation/degradation of A-chain in the cytosol. The likely implications of these findings to the survival of the toxin-producing tissue are discussed.
Jolliffe
Nicholas A
NA
Department of Biological Sciences, University of Warwick, Coventry CV4 7AL, United Kingdom.
Di Cola
Alessandra
A
Marsden
Catherine J
CJ
Lord
J Michael
JM
Ceriotti
Aldo
A
Frigerio
Lorenzo
L
Roberts
Lynne M
LM
D000595:Amino Acid Sequence; D001692:Biological Transport / Q000235:genetics; D002367:Ricinus communis; D004721:Endoplasmic Reticulum / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D006031:Glycosylation; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000737:chemistry* / Q000235:genetics / Q000502:physiology; D011498:Protein Precursors / Q000737:chemistry* / Q000235:genetics / Q000502:physiology; D021122:Protein Subunits / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D011523:Protoplasts / Q000737:chemistry / Q000378:metabolism*; D012276:Ricin / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D014026:Nicotiana / Q000737:chemistry / Q000166:cytology / Q000378:metabolism*
D010446:Peptide Fragments; D011498:Protein Precursors; D021122:Protein Subunits; D012276:Ricin