Proteomics Analysis Identifies New Components of the Fission and Budding Yeast Anaphase-Promoting Complexes

Yoon, Hyun-Joo; Feoktistova, Anna; Wolfe, Benjamin; Jennings, Jennifer L.; Link, Andrew J.; Gould, Kathleen L.

doi:10.1016/s0960-9822(02)01331-3

Cited by 97 publications

(99 citation statements)

References 31 publications

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“…Protein complexes were obtained using TAP strategy as described (Tasto et al, 2001) with one variation: the lysates were clarified at 3000 rpm on a tabletop GS-6R centrifuge in lieu of ultracentrifugation. TAP pellets were subjected to mass spectrometric analyses as described (Yoon et al, 2002).…”

Section: Protein Methodsmentioning

confidence: 99%

“…Both the alp4-TAP and alp6-TAP strains grew normally, suggesting that the epitope did not compromise the function of either protein. Tandem affinity purification steps were then carried out from both strains and the protein composition of a portion of each TAP complex was analyzed by silver staining (unpublished data) with the remainder analyzed by DALPC tandem mass spectrometry (Yoon et al, 2002). Proteins identified from both purifications at greater than 10% sequence coverage that were absent from the complex purified from untagged cells or from unrelated TAP purifications (unpublished data) are listed in Table 2.…”

Section: Purification Of S Pombe ␥-Tuc-associated Proteinsmentioning

confidence: 99%

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Identification and Characterization of Two Novel Proteins Affecting Fission Yeast γ-tubulin Complex Function

Venkatram

Tasto

Feoktistova

et al. 2004

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The ␥-tubulin complex, via its ability to organize microtubules, is critical for accurate chromosome segregation and cytokinesis in the fission yeast, Schizosaccharomyces pombe. To better understand its roles, we have purified the S. pombe ␥-tubulin complex. Mass spectrometric analyses of the purified complex revealed known components and identified two novel proteins (i.e., Mbo1p and Gfh1p) with homology to ␥-tubulin-associated proteins from other organisms. We show that both Mbo1p and Gfh1p localize to microtubule organizing centers. Although cells deleted for either mbo1 ؉ or gfh1 ؉ are viable, they exhibit a number of defects associated with altered microtubule function such as defects in cell polarity, nuclear positioning, spindle orientation, and cleavage site specification. In addition, mbo1⌬ and gfh1⌬ cells exhibit defects in astral microtubule formation and anchoring, suggesting that these proteins have specific roles in astral microtubule function. This study expands the known roles of ␥-tubulin complex components in organizing different types of microtubule structures in S. pombe.

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Section: Protein Methodsmentioning

confidence: 99%

Section: Purification Of S Pombe ␥-Tuc-associated Proteinsmentioning

confidence: 99%

Identification and Characterization of Two Novel Proteins Affecting Fission Yeast γ-tubulin Complex Function

Venkatram

Tasto

Feoktistova

et al. 2004

MBoC

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114

191

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“…The APC is a large protein complex containing at least 11 core subunits (Zachariae et al, 1998b;Gmachl et al, 2000;Yoon et al, 2002) that can further associate with at least three known different activators (Fang et al, 1998a, b;Kallio et al, 1998;Zachariae et al, 1998a;Cooper et al, 2000). The majority of these subunits are stably associated throughout the cell cycle (Peters et al, 1996;Grossberger et al, 1999) except for the different activators whose binding to APC is cell cycle regulated (Fang et al, 1998b;Zachariae et al, 1998a;Cooper et al, 2000).…”

Section: Apc Composition and Structurementioning

confidence: 99%

The anaphase-promoting complex: a key factor in the regulation of cell cycle

et al. 2005

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Events controlling cell division are governed by the degradation of different regulatory proteins by the ubiquitin-dependent pathway. In this pathway, the attachment of a polyubiquitin chain to a substrate by an ubiquitin-ligase targets this substrate for degradation by the 26S proteasome. Two different ubiquitin ligases play an important role in the cell cycle: the SCF (Skp1/Cullin/ F-box) and the anaphase-promoting complex (APC). In this review, we describe the present knowledge about the APC. We pay particular attention to the latest results concerning APC structure, APC regulation and substrate recognition, and we discuss the implication of these findings in the understanding the APC function.

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“…For the Cdk1 kinase assay, Ϸ50 ng of recombinant Cdk1 kinase complex, purified from baculovirus-infected insect cells as previously described (Yoon et al, 2002), was incubated with 200 ng of bacterially produced His6-Nbl1(1-95) in a reaction buffer containing 50 mM Tris pH 7.4, 10 mM MgCl 2 , 2 M DTT, 10 M unlabeled ATP, and 5 Ci of [␥-32 P] ATP. After incubation with shaking at 30°C for 30 min, reactions were terminated with 5ϫ sample buffer, and samples were boiled and separated by SDS-PAGE.…”

Section: In Vitro Kinase Assaysmentioning

confidence: 99%

“…The gel was cut in half and the upper portion (30 to 90 kDa) was transferred to a PVDF membrane and blotted with anti-GFP antibodies to detect Ark1-GFP. The lower part (10 to 30 kDa) of the same gel was stained by Coomassie blue dye to visualize Histone H3.2, and, after drying of the gel, autoradiography was used to check incorporation of 32 P. For the Cdk1 kinase assay, Ϸ50 ng of recombinant Cdk1 kinase complex, purified from baculovirus-infected insect cells as previously described (Yoon et al, 2002), was incubated with 200 ng of bacterially produced His6-Nbl1(1-95) in a reaction buffer containing 50 mM Tris pH 7.4, 10 mM MgCl 2 , 2 M DTT, 10 M unlabeled ATP, and 5 Ci of [␥-32 P] ATP. After incubation with shaking at 30°C for 30 min, reactions were terminated with 5ϫ sample buffer, and samples were boiled and separated by SDS-PAGE.…”

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confidence: 99%

A Link between Aurora Kinase and Clp1/Cdc14 Regulation Uncovered by the Identification of a Fission Yeast Borealin-Like Protein

Bohnert

Chen

Clifford

et al. 2009

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The chromosomal passenger complex (CPC) regulates various events in cell division. This complex is composed of a catalytic subunit, Aurora B kinase, and three nonenzymatic subunits, INCENP, Survivin, and Borealin. Together, these four subunits interdependently regulate CPC function, and they are highly conserved among eukaryotes. However, a Borealin homologue has never been characterized in the fission yeast, Schizosaccharomyces pombe. Here, we isolate a previously uncharacterized S. pombe protein through association with the Cdc14 phosphatase homologue, Clp1/Flp1, and identify it as a Borealin-like member of the CPC. Nbl1 (novel Borealin-like 1) physically associates with known CPC components, affects the kinase activity and stability of the S. pombe Aurora B homologue, Ark1, colocalizes with known CPC subunits during mitosis, and shows sequence similarity to human Borealin. Further analysis of the Clp1-Nbl1 interaction indicates that Clp1 requires CPC activity for proper accumulation at the contractile ring (CR). Consistent with this, we describe negative genetic interactions between mutant alleles of CPC and CR components. Thus, this study characterizes a fission yeast Borealin homologue and reveals a previously unrecognized connection between the CPC and the process of cytokinesis in S. pombe. INTRODUCTIONTo ensure successful cell division, sister chromatids must segregate to opposite poles of dividing cells in mitosis and be partitioned into two new daughter cells during cytokinesis (Pines and Rieder, 2001). Highly intricate mechanisms control these processes, with various macromolecular complexes coordinating their activities such that the integrity of cell division is maintained. The chromosomal passenger complex (CPC), which is composed of a catalytic subunit, Aurora B kinase, and three nonenzymatic subunits, INCENP, Survivin, and Borealin, functions as one of these critical regulatory complexes. As its name implies, this complex travels on chromosomes to various sites during cell division such that it can execute specific tasks at distinct locations and times (Earnshaw and Bernat, 1991). These functions include, but are not limited to, chromosome condensation, stabilization of the mitotic spindle, correction of improper kinetochore-microtubule attachments, and regulation of cytokinesis (for reviews, see Vagnarelli and Earnshaw, 2004;Vader et al., 2006;Ruchaud et al., 2007).The four CPC subunits are highly interdependent for proper localization, activity, and stability (Ruchaud et al., 2007). Consistent with such interdependency, these subunits are widely conserved among eukaryotes (Ruchaud et al., 2007). However, it has proven difficult to identify Borealin homologues in yeasts. Though some have speculated that yeast Borealin homologues might not exist because of fusion of the Borealin and Survivin subunits into a single yeast Survivin homologue (Vader et al., 2006), recent evidence suggests that there are in fact yeast Borealin homologues (Nakajima et al., 2009). Nonetheless, a Borealin homologue in th...

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Proteomics Analysis Identifies New Components of the Fission and Budding Yeast Anaphase-Promoting Complexes

Cited by 97 publications

References 31 publications

Identification and Characterization of Two Novel Proteins Affecting Fission Yeast γ-tubulin Complex Function

Identification and Characterization of Two Novel Proteins Affecting Fission Yeast γ-tubulin Complex Function

The anaphase-promoting complex: a key factor in the regulation of cell cycle

A Link between Aurora Kinase and Clp1/Cdc14 Regulation Uncovered by the Identification of a Fission Yeast Borealin-Like Protein

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