Peroxisomes are single membrane bound compartments. They are thought to be present in almost all eukaryotic cells, although the bulk of our knowledge about peroxisomes has been generated from only a handful of model organisms. Peroxisomal matrix proteins are synthesized cytosolically and posttranslationally imported into the peroxisomal matrix. The import is generally thought to be mediated by two different targeting signals. These are respectively recognized by the two import receptor proteins Pex5 and Pex7, which facilitate transport across the peroxisomal membrane.
Here, we show the first in vivo localization studies of peroxisomes in a representative organism of the ecologically relevant group of diatoms using fluorescence and transmission electron microscopy. By expression of various homologous and heterologous fusion proteins we demonstrate that targeting of Phaeodactylum tricornutum peroxisomal matrix proteins is mediated only by PTS1 targeting signals, also for proteins that are in other systems imported via a PTS2 mode of action. Additional in silico analyses suggest this surprising finding may also apply to further diatoms. Our data suggest that loss of the PTS2 peroxisomal import signal is not reserved to Caenorhabditis elegans as a single exception, but has also occurred in evolutionary divergent organisms. Obviously, targeting switching from PTS2 to PTS1 across different major eukaryotic groups might have occurred for different reasons. Thus, our findings question the widespread assumption that import of peroxisomal matrix proteins is generally mediated by two different targeting signals. Our results implicate that there apparently must have been an event causing the loss of one targeting signal even in the group of diatoms. Different possibilities are discussed that indicate multiple reasons for the detected targeting switching from PTS2 to PTS1.
P. tricornutum cells expressing different types of GFP fusion proteins were harvested via centrifugation at 1,500 × g and cryoimmobilized by high-pressure freezing on gold carriers.
Figure: (A) Ultrathin section of P. tricornutum expressing Pex10-GFP in Epon without antibody labeling. The boxed area is shown in (B) at higher magnification and illustrates two peroxisomes in proximity to the nucleus, the golgi and the plastid.
CW, cell wall; G, golgi apparatus; Mt, mitochondrium; Ne, nuclear envelope; Nu, nucleus; P, peroxisome; Pl, plastid; V, vacuole. Scale bars represent 2 μm (A), 500 nm (B).