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Mah AS, et al.  (2005) Substrate specificity analysis of protein kinase complex Dbf2-Mob1 by peptide library and proteome array screening. BMC Biochem 6():22

Abstract: BACKGROUND: The mitotic exit network (MEN) is a group of proteins that form a signaling cascade that is essential for cells to exit mitosis in Saccharomyces cerevisiae. The MEN has also been implicated in playing a role in cytokinesis. Two components of this signaling pathway are the protein kinase Dbf2 and its binding partner essential for its kinase activity, Mob1. The components of MEN that act upstream of Dbf2-Mob1 have been characterized, but physiological substrates for Dbf2-Mob1 have yet to be identified. RESULTS: Using a combination of peptide library selection, phosphorylation of opitmal peptide variants, and screening of a phosphosite array, we found that Dbf2-Mob1 preferentially phosphorylated serine over threonine and required an arginine three residues upstream of the phosphorylated serine in its substrate. This requirement for arginine in peptide substrates could not be substituted with the similarly charged lysine. This specificity determined for peptide substrates was also evident in many of the proteins phosphorylated by Dbf2-Mob1 in a proteome chip analysis. CONCLUSIONS: We have determined by peptide library selection and phosphosite array screening that the protein kinase Dbf2-Mob1 preferentially phosphorylated substrates that contain an RXXS motif. A subsequent proteome microarray screen revealed proteins that can be phosphorylated by Dbf2-Mob1 in vitro. These proteins are enriched for RXXS motifs, and may include substrates that mediate the function of Dbf2-Mob1 in mitotic exit and cytokinesis. The relatively low degree of sequence restriction at the site of phosphorylation suggests that Dbf2 achieves specificity by docking its substrates at a site that is distinct from the phosphorylation site.

Status: Published Type: Journal Article PubMed ID: 16242037

Topics addressed in this paper

Number of different genes curated to this paper: 69

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Topics Topics not linked to Genes Genes linked to topics (#1 - 10 )
ABF2 ADD37 ADH7 ADK1 AIM17 AIM3 AMD1 ARC1 AVT3 AVT4
Additional Literature blue ball blue ball blue ball blue ball blue ball blue ball blue ball blue ball blue ball blue ball
Large-scale protein modification yg ball
Omics yg ball
Protein Processing/Modification/Regulation blue ball blue ball blue ball blue ball blue ball blue ball blue ball blue ball blue ball blue ball
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Topics Genes linked to topics (#11 - 20 )
BDF2 BFR2 CBF1 CDC13 DBF2 DOT5 DRS1 ESBP6 FPK1 GUS1
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Function/Process blue ball
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Topics Genes linked to topics (#21 - 30 )
HSP42 HSV2 HTZ1 KKQ8 MES1 MET17 MOB1 MRPL9 NCL1 NDJ1
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Function/Process blue ball
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Topics Genes linked to topics (#31 - 40 )
NET1 NHP10 NIP7 NSE1 NTR2 NVJ2 OAF1 OSH2 PAH1 PRE10
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Topics Genes linked to topics (#41 - 50 )
PRM10 QCR6 RNT1 RPL43B RRP5 SIS1 SRO77 TEF2 TGL1 THI73
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Topics Genes linked to topics (#51 - 60 )
VHS2 VPS27 YBR138C YBR285W YDR134C YGR038C-A YHR182W YJL211C YJL213W YJR142W
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Topics Genes linked to topics (#61 - 69 )
YKL077W YLR177W YML037C YMR196W YNL155W YNL284C-A YOR228C YPL150W YPL257W-A
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