Identification of a novel mitotic phosphorylation motif associated with protein localization to the mitotic apparatus

Journal Article · Fri Nov 16 00:00:00 EST 2007 · Journal of Cell Science, 120(22):4060-70

DOI:https://doi.org/10.1242/jcs.014795· OSTI ID:921417

Yang, Feng; Camp, David G; Gritsenko, Marina A; Luo, Quanzhou; Kelly, Ryan T; Clauss, Therese RW; Brinkley, William R; Smith, Richard D; Stenoien, David L

The chromosomal passenger complex (CPC) is a critical regulator of chromosome, cytoskeleton and membrane dynamics during mitosis. Here, we identified phosphopeptides and phosphoprotein complexes recognized by a phosphorylation specific antibody that labels the CPC using liquid chromatography coupled to mass spectrometry. A mitotic phosphorylation motif (PX{G/T/S}{L/M}[pS]P or WGL[pS]P) was identified in 11 proteins including Fzr/Cdh1 and RIC-8, two proteins with potential links to the CPC. Phosphoprotein complexes contained known CPC components INCENP, Aurora-B and TD-60, as well as SMAD2, 14-3-3 proteins, PP2A, and Cdk1, a likely kinase for this motif. Protein sequence analysis identified phosphorylation motifs in additional proteins including SMAD2, Plk3 and INCENP. Mitotic SMAD2 and Plk3 phosphorylation was confirmed using phosphorylation specific antibodies, and in the case of Plk3, phosphorylation correlates with its localization to the mitotic apparatus. A mutagenesis approach was used to show INCENP phosphorylation is required for midbody localization. These results provide evidence for a shared phosphorylation event that regulates localization of critical proteins during mitosis.

• Research Organization: Pacific Northwest National Lab. (PNNL), Richland, WA (United States). Environmental Molecular Sciences Lab. (EMSL)
• Sponsoring Organization: USDOE
• DOE Contract Number: AC05-76RL01830
• OSTI ID: 921417
• Report Number(s): PNNL-SA-55602; JNCSAI; 12294; TRN: US200804%%805
• Journal Information: Journal of Cell Science, 120(22):4060-70, Vol. 120, Issue 22; ISSN 0021-9533
• Country of Publication: United States
• Language: English

Related Subjects

59 BASIC BIOLOGICAL SCIENCES
60 APPLIED LIFE SCIENCES
ANTIBODIES
CHROMATOGRAPHY
MASS SPECTROSCOPY
MEMBRANES
MITOSIS
MUTAGENESIS
OCCUPANTS
PHOSPHOPROTEINS
PHOSPHORYLATION
PHOSPHOTRANSFERASES
PROTEINS
STRUCTURAL CHEMICAL ANALYSIS
Phosphorylation
Mitosis
Proteomics
Chromosomal Passenger Complex
Environmental Molecular Sciences Laboratory