Dataset Information

Signaling pathways altered at the mitotic spindle in clathrin depleted cells

ABSTRACT: We present a snapshot of the regulation of mitotic-spindle proteins and their phosphorylation status in the absence of clathrin during metaphase. A reference list of proteins and phosphosites identified from a mitotic spindle preparation was generated using a label-free and a parallel chemical dimethyl isotopic peptide labelling approach. We further designed experiments to analyse proteins and phosphosites at the mitotic spindle whose abundance was affected by the knock-down of clathrin. We used dimethylation labelling of peptides in combination with fractionation strategies to identify and quantify phosphopeptides enriched by TiO2 beads and the flow-through non-phoshopeptides by ESI-LC-MS/MS for a broad coverage of the mitotic spindle proteome in the absence of clathrin.

INSTRUMENT(S): LTQ Orbitrap Velos

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Permanent Cell Line Cell

DISEASE(S): Cancer

SUBMITTER: Sushma Rao

LAB HEAD: A/Prof. Megan Chircop

PROVIDER: PXD001603 | Pride | 2016-05-17

REPOSITORIES: Pride

ACCESS DATA

Dataset's files

Source:

			Action	DRS
	Newruncombinedfractions.zip	Other
	Nonphosphocomblongcolumn.zip	Other
	Nonphosphofrac18cmcolumn.zip	Other
	Nonphosphofrac18cmcolumntxtfiles.zip	Other
	Nonphospholongcolmntxtfiles.zip	Other

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Publications

The Clathrin-dependent Spindle Proteome.

Rao Sushma R SR Flores-Rodriguez Neftali N Page Scott L SL Wong Chin C Robinson Phillip J PJ Chircop Megan M

Molecular & cellular proteomics : MCP 20160512 8

The mitotic spindle is required for chromosome congression and subsequent equal segregation of sister chromatids. These processes involve a complex network of signaling molecules located at the spindle. The endocytic protein, clathrin, has a "moonlighting" role during mitosis, whereby it stabilizes the mitotic spindle. The signaling pathways that clathrin participates in to achieve mitotic spindle stability are unknown. Here, we assessed the mitotic spindle proteome and phosphoproteome in clathr ...[more]

PMID: 27174698

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