Phosphorylation of SYP-5 S541 requires both PLK-1 and PLK-2, but not the PLK-docking site on SYP-1. (A) Schematic showing in vitro PLK-2 phosphorylation sites on SYP-5, with sequence coverage from mass spectrometry analysis indicated by gray blocks. Conserved residues that conform to the PLK consensus motif are highlighted in magenta. (B) In vitro kinase assays using recombinant 6His-MBP-SYP-5 incubated with or without PLK-2. Coomassie staining of the purified proteins (left) and western blot for SYP-5 pS541 (right) are shown. 6His-MBP-SYP-5 is indicated by an arrow. (C) Immunofluorescence images of pachytene nuclei from wild type, plk-2(tm1395), plk-1(RNAi), plk-2(tm1395); plk-1(RNAi), and syp-1T452A animals, stained for SYP-1 (magenta), and SYP-5 pS541 (green). Scale bar, 5 µm. Source data are available for this figure: SourceData F2.
Figure 2.

Phosphorylation of SYP-5 S541 requires both PLK-1 and PLK-2, but not the PLK-docking site on SYP-1. (A) Schematic showing in vitro PLK-2 phosphorylation sites on SYP-5, with sequence coverage from mass spectrometry analysis indicated by gray blocks. Conserved residues that conform to the PLK consensus motif are highlighted in magenta. (B) In vitro kinase assays using recombinant 6His-MBP-SYP-5 incubated with or without PLK-2. Coomassie staining of the purified proteins (left) and western blot for SYP-5 pS541 (right) are shown. 6His-MBP-SYP-5 is indicated by an arrow. (C) Immunofluorescence images of pachytene nuclei from wild type, plk-2(tm1395), plk-1(RNAi), plk-2(tm1395); plk-1(RNAi), and syp-1T452A animals, stained for SYP-1 (magenta), and SYP-5 pS541 (green). Scale bar, 5 µm. Source data are available for this figure: SourceData F2.

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