Centrosomes at M phase act as a scaffold for the accumulation of intracellular ubiquitinated proteins.

Abstract

Centrosome size varies considerably during the cell cycle; it is greatest during metaphase, partly because of pericentriolar matrix recruitment and an increase in microtubule-organizing activity. However, the mechanism of centrosome maturation during M phase is poorly defined. In the present study, we identified and quantified centrosomal proteins during S and M phases using stable isotope labeling by amino acids in cell culture (SILAC) coupled with liquid chromatography-tandem mass spectrometry (LC-MS/MS). We identified 991 proteins, of which 310 and 325 proteins were upregulated during S and M phases, respectively. Ubiquitinated proteins containing K48- and K63-linked polyubiquitin chains accumulated in the centrosomes during M phase, although 26S proteasome activity in the centrosomes did not markedly differ between S and M phases. Conversely, cytoplasmic dynein, which transports ubiquitinated proteins to the centrosomes, increased 2-fold in the centrosomes during M phase relative to S phase. Furthermore, PYR-41, a ubiquitin E1 inhibitor, reduced centrosome size during metaphase, causing increased aneuploidy. RNA interference suppression of Ecm29, which inhibits proteasome activity, decreased the accumulation of ubiquitinated proteins in the centrosomes. These results show that accumulation of ubiquitinated proteins promotes centrosome maturation during M phase and further suggest a novel function of centrosomes as a scaffold temporarily gathering intracellular ubiquitinated proteins.

• Authors: Hitomi Kimura, Yoshio Miki, Akira Nakanishi
• Journal: Cell cycle (Georgetown, Tex.) (Cell Cycle) Vol. 13 Issue 12 Pg. 1928-37 ( 2014) ISSN: 1551-4005 [Electronic] United States
• PMID: 24743317 (Publication Type: Journal Article)

Chemical References

• Ubiquitinated Proteins
• Polyubiquitin

Topics

• Cell Division (physiology)
• Cell Line, Tumor
• Centrosome (metabolism)
• Humans
• Polyubiquitin (metabolism)
• S Phase (physiology)
• Ubiquitinated Proteins (metabolism)