We have studied the subcellular localization of the
acid S-like
ribonuclease (
RNase) LX in tomato (Lycopersicon esculentum Mill.) cells using a combination of biochemical and immunological methods. It was found that the
enzyme, unexpectedly excluded from highly purified vacuoles, accumulates in the endoplasmic reticulum. The evidence that
RNase LX is a resident of the endoplasmic reticulum (ER) is supported by an independent approach showing that the C-terminal
peptide HDEF of
RNase LX acts as an alternative ER retention signal in plants. For functional testing, the cellular distribution of chimeric
protein constructs based on a marker
protein, Brazil nut (Bertholletia excelsa) 2S
albumin, was analyzed immunochemically in transgenic tobacco (Nicotiana tabacum) plants. Here, we report that the
peptide motif is necessary and sufficient to accumulate 2S
albumin constructs of both vacuolar and extracellular final destinations in the ER. We have shown immunochemically that
RNase LX is specifically expressed during endosperm mobilization and leaf and flower senescence. Using immunofluorescence,
RNase LX protein was detected in immature tracheary elements, suggesting a function in xylem differentiation. These results support a physiological function of
RNase LX in selective cell death processes that are also thought to involve programmed cell death. It is assumed that
RNase LX accumulates in an ER-derived compartment and is released by membrane disruption into the cytoplasma of those cells that are intended to undergo
autolysis. These processes are accompanied by degradation of cellular components supporting a metabolic recycling function of the intracellular
RNase LX.