Pericentrin and γ-Tubulin Form a Protein Complex and Are Organized into a Novel Lattice at the Centrosome

Dictenberg, Jason B.; Zimmerman, Wendy; Sparks, Cynthia A.; Young, Aaron Isadore; Vidair, Charles; Zheng, Yixian; Carrington, Walter A.; Fay, Fredric S.; Doxsey, Stephen

doi:10.1083/jcb.141.1.163

Cited by 463 publications

(402 citation statements)

References 47 publications

Supporting

Mentioning

384

Contrasting

Unclassified

Order By: Relevance

“…However, γ-tubulin localization to pericentrindeficient centrosomes was reduced, as has been seen in other models. 4,5,7,18 We also observed a significant increase in the proportion of monopolar spindles, but not to the extent observed in knockdown experiments in HeLa cells. 7,18 Knockdown of HeLa PCNT reduced the centrosomal γ-tubulin to approximately 10% of wild-type levels, 18 a more dramatic reduction than we observed and one that could impact sufficiently on centrosome maturation to lead to monopolar spindle formation in almost all cells.…”

Section: Discussionmentioning

confidence: 40%

See 1 more Smart Citation

Promoter hijack reveals pericentrin functions in mitosis and the DNA damage response

et al. 2013

View full text Add to dashboard Cite

Section: Discussionmentioning

confidence: 40%

“…[1][2][3][4] Pericentrin acts as scaffold within the PCM and interacts with a number of partners, in particular γ-tubulin and other members of the γ-tubulin ring complex, providing a structural framework in which the MTOC activities can occur [5][6][7] (reviewed in ref. 8).…”

Section: Introductionmentioning

confidence: 99%

Promoter hijack reveals pericentrin functions in mitosis and the DNA damage response

et al. 2013

View full text Add to dashboard Cite

“…In contrast, the pericentriolar material is structurally less well defined and has often been described as ''amorphous'' or ''electron dense'' material. Recently however, electron microscopy tomography and high-resolution fluorescence microscopy of centrosomes from Drosophila, Spisula and mammalian cells have identified ring-like structures and protein complexes of higher order (Dictenberg et al 1998;Moritz et al 1995a;Vogel et al 1997). These ring-like structures colocalize with gamma-tubulin, a conserved centrosomal component, in immunoelectron microscopy (Moritz et al 1995b) suggesting them to be akin to the c-tubulin ring complex (cTuRC) involved in microtubule nucleation.…”

Section: Introductionmentioning

confidence: 99%

Structure and microtubule-nucleation activity of isolated Drosophila embryo centrosomes characterized by whole mount scanning and transmission electron microscopy

Lange

Kirfel

Gestmann

et al. 2005

Histochem Cell Biol

View full text Add to dashboard Cite

Experimental approaches in Drosophila melanogaster over the last 20 years have played a fundamental role in elucidating the function, structure and molecular composition of the centrosome. However, quantitative data on the structure and function of the Drosophila centrosome are still lacking. This study uses, for the first time, whole mount electron microscopy in combination with negative staining on isolated centrosomes from the early Drosophila embryos to analyze its dimensions, structure and capacity to nucleate microtubules in vitro. We show that these organelles are on average 0.75 lm in diameter and have abundant pericentriolar material which often appears fibrillar and with bulbous protrusions. Corresponding to the abundant pericentriolar material, extensive microtubule nucleation occurs. Quantification of the number of microtubules nucleated showed that 50-300 active nucleation sites are present. We examined via electron microscopy immunogold labeling the distribution of c-tubulin, CNN, Asp and the MPM-2 epitopes that are phosphorylated through Polo and the Cdk1 kinase. The distribution of these proteins is homogeneous, with the MPM-2 epitopes exhibiting the highest density. In contrast, centrosomal subdomains are identified using a centriole marker to relate centrosome size to the centriole number by electron microscopy. In conclusion, we present a clear-cut technique assaying and quantifying the microtubule nucleation capacity and antigen distribution complementing molecular studies on centrosome protein complexes, cell organelle assembly and protein composition.

show abstract

“…M30 Cytodeath and anti-HA rat monoclonal antibody 3F10 was obtained from Roche Diagnostics (Indianapolis, IN) Anti-human lamin A/C antibody was purchased from Cell Signaling Technology (Beverley, MA). Other antibodies included M8 anti-pericentrin antibody, (Dictenberg et al, 1998), human autoimmune serum 5051 that recognizes centrosome proteins (Doxsey et.al., 1994), anti-pericentrin B/kendrin-specific antibody (Flory et.al., 2000) (obtained from Trisha Davis, University of Washington, Seattle, WA), anti-GCP2 antibody (Murphy et al, 1998) (obtained from Dr. Tim Stearns), and anti-GCP3 antibody (a gift from Michel Bornens, Institut Curie, Paris, France). …”

mentioning

confidence: 99%

Mitosis-specific Anchoring of γ Tubulin Complexes by Pericentrin Controls Spindle Organization and Mitotic Entry

Zimmerman

Sillibourne

Rosa

et al. 2004

MBoC

Self Cite

275

258

View full text Add to dashboard Cite

Microtubule nucleation is the best known function of centrosomes. Centrosomal microtubule nucleation is mediated primarily by ␥ tubulin ring complexes (␥ TuRCs). However, little is known about the molecules that anchor these complexes to centrosomes. In this study, we show that the centrosomal coiled-coil protein pericentrin anchors ␥ TuRCs at spindle poles through an interaction with ␥ tubulin complex proteins 2 and 3 (GCP2/3). Pericentrin silencing by small interfering RNAs in somatic cells disrupted ␥ tubulin localization and spindle organization in mitosis but had no effect on ␥ tubulin localization or microtubule organization in interphase cells. Similarly, overexpression of the GCP2/3 binding domain of pericentrin disrupted the endogenous pericentrin-␥ TuRC interaction and perturbed astral microtubules and spindle bipolarity. When added to Xenopus mitotic extracts, this domain uncoupled ␥ TuRCs from centrosomes, inhibited microtubule aster assembly, and induced rapid disassembly of preassembled asters. All phenotypes were significantly reduced in a pericentrin mutant with diminished GCP2/3 binding and were specific for mitotic centrosomal asters as we observed little effect on interphase asters or on asters assembled by the Ran-mediated centrosome-independent pathway. Additionally, pericentrin silencing or overexpression induced G2/antephase arrest followed by apoptosis in many but not all cell types. We conclude that pericentrin anchoring of ␥ tubulin complexes at centrosomes in mitotic cells is required for proper spindle organization and that loss of this anchoring mechanism elicits a checkpoint response that prevents mitotic entry and triggers apoptotic cell death. INTRODUCTIONThe centrosome is the primary microtubule-organizing center in animal cells. At the centrosome core is a pair of barrel-shaped microtubule assemblies, the centrioles (Doxsey, 2001). Centrioles are capable of self-assembly (Marshall et al., 2001;Khodjakov et al., 2002) and can serve as templates for recruitment and organization of the surrounding pericentriolar matrix (Bobinnec et al., 1998;Kirkham et al., 2003). The pericentriolar material or centrosome matrix contains a high proportion of coiled coil proteins and is the site of microtubule nucleation. Within the matrix are large protein complexes of ␥ tubulin and associated proteins that have a ring-like structure and mediate the nucleation of microtubules called ␥ tubulin ring complexes or ␥ TuRCs (Moritz et al., 1995a;Zheng et al., 1995). Other proteins may share the ability to nucleate microtubules because centrosomes can organize microtubules in the absence of functional ␥ tubulin (Sampaio et al., 2001;Strome et al., 2001;Hannak et al., 2002).During cell cycle progression, centrosomes "mature" by recruiting additional ␥ TuRCs and several other proteins, resulting in an increase in the nucleation capacity of the centrosome (reviewed in Blagden and Glover, 2003). However, we still know very little about proteins that directly anchor ␥ TuRCs to centrosomes in vertebrate cells. ...

show abstract

Pericentrin and γ-Tubulin Form a Protein Complex and Are Organized into a Novel Lattice at the Centrosome

Cited by 463 publications

References 47 publications

Promoter hijack reveals pericentrin functions in mitosis and the DNA damage response

Promoter hijack reveals pericentrin functions in mitosis and the DNA damage response

Structure and microtubule-nucleation activity of isolated Drosophila embryo centrosomes characterized by whole mount scanning and transmission electron microscopy

Mitosis-specific Anchoring of γ Tubulin Complexes by Pericentrin Controls Spindle Organization and Mitotic Entry

Contact Info

Product

Resources

About