Microtubule Nucleation at Non-Spindle Pole Body Microtubule-Organizing Centers Requires Fission Yeast Centrosomin-Related Protein mod20p

Sawin, Kenneth E.; Lourenco, Paula C.; Snaith, Hilary A.

doi:10.1016/j.cub.2004.03.042

Cited by 173 publications

(301 citation statements)

References 58 publications

Supporting

Mentioning

274

Contrasting

Order By: Relevance

“…ϩ , under control of the nmt81 promoter (Sawin et al, 2004) and Sid4-CFP. Many cells (24%) were observed with greater-than-normal numbers of cytoplasmic MTs emanating from the SPBs ( Figure 7D).…”

Section: Ppc89 Overproduction Is Lethal and Results In Spb Enlargementmentioning

confidence: 99%

“…Antibodies 12CA5 and 9E10 were used at 2 g/ml in Tris-buffered saline to detect epitope-tagged proteins. These antibodies were then detected b Constructed by crosses among other strains listed here or between such strains and strains described by West et al (1998), Chang and Gould (2000), Flory et al (2002), Tomlin et al (2002), Sawin et al (2004), and Venkatram et al (2004).…”

Section: Protein Methodsmentioning

confidence: 99%

See 1 more Smart Citation

Ppc89 Links Multiple Proteins, Including the Septation Initiation Network, to the Core of the Fission Yeast Spindle-Pole Body

Rosenberg

Tomlin

McDonald

et al. 2006

MBoC

View full text Add to dashboard Cite

The spindle-pole body (SPB), the yeast analog of the centrosome, serves as the major microtubule (MT) organizing center in the yeast cell. In addition to this central function, the SPB organizes and concentrates proteins required for proper coordination between the nuclear-division cycle and cytokinesis. For example, the Schizosaccharomyces pombe septationinitiation network (SIN), which is responsible for initiating actomyosin ring constriction and septation, is assembled at the SPB through its two scaffolding components, Sid4 and Cdc11. In an effort to identify novel SIN interactors, we purified Cdc11 and identified by mass spectrometry a previously uncharacterized protein associated with it, Ppc89. Ppc89 localizes constitutively to the SPB and interacts directly with Sid4. A fusion between the N-terminal 300 amino acids of Sid4 and a SPB targeting domain of Ppc89 supplies the essential function of Sid4 in anchoring the SIN. ppc89⌬ cells are inviable and exhibit defects in SPB integrity, and hence in spindle formation, chromosome segregation, and SIN localization. Ppc89 overproduction is lethal, resulting primarily in a G2 arrest accompanied by massive enlargement of the SPB and increased SPB MT nucleation. These results suggest a fundamental role for Ppc89 in organization of the S. pombe SPB.

show abstract

Section: Ppc89 Overproduction Is Lethal and Results In Spb Enlargementmentioning

confidence: 99%

Section: Protein Methodsmentioning

confidence: 99%

Ppc89 Links Multiple Proteins, Including the Septation Initiation Network, to the Core of the Fission Yeast Spindle-Pole Body

Rosenberg

Tomlin

McDonald

et al. 2006

MBoC

View full text Add to dashboard Cite

show abstract

“…Translocation of ␥-TuCs on MTs is also observed in cells lacking mto1p, a centrosomin-related protein involved in the recruitment of ␥-TuCs to iMTOCs (Sawin et al, 2004;Zimmerman and Chang, 2005). Studies of mto2p, to which mto1p binds directly (Samejima et al, 2005), provide further evidence that nucleating components are transported along pre-existing MTs in S. pombe (Janson et al, 2005).…”

Section: Generation Of Noncentrosomal Mtsmentioning

confidence: 90%

Generation of noncentrosomal microtubule arrays

Bartolini

Gundersen

2006

Journal of Cell Science

228

219

View full text Add to dashboard Cite

In most proliferating and migrating animal cells, the centrosome is the main site for microtubule (MT) nucleation and anchoring, leading to the formation of radial MT arrays in which MT minus ends are anchored at the centrosomes and plus ends extend to the cell periphery. By contrast, in most differentiated animal cell types, including muscle, epithelial and neuronal cells, as well as most fungi and vascular plant cells, MTs are arranged in noncentrosomal arrays that are non-radial. Recent studies suggest that these noncentrosomal MT arrays are generated by a three step process. The initial step involves formation of noncentrosomal MTs by distinct mechanisms depending on cell type: release from the centrosome, catalyzed nucleation at noncentrosomal sites or breakage of pre-existing MTs. The second step involves transport by MT motor proteins or treadmilling to sites of assembly. In the final step, the noncentrosomal MTs are rearranged into cell-type-specific arrays by bundling and/or capture at cortical sites, during which MTs acquire stability. Despite their relative stability, the final noncentrosomal MT arrays may still exhibit dynamic properties and in many cases can be remodeled.

show abstract

“…4 A and C). We performed collocation studies using additional fluorescent markers [ase1-3xmCherry for the MT overlap zone; mod20-GFP for the MT organization center (MTOC)] (29,30), and found that these patches resided in the vicinity of MTOC in the MT overlap zone (Fig. S3 C and D).…”

Section: Identification Of Discrete Dam1 Assemblies On Cytoplasmic Mtmentioning

confidence: 99%

A non-ring-like form of the Dam1 complex modulates microtubule dynamics in fission yeast

Gao

Courthéoux

Gachet

et al. 2010

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

The Dam1 complex is a kinetochore component that couples chromosomes to the dynamic ends of kinetochore microtubules (kMTs). Work in the budding yeast Saccharomyces cerevisiae has shown that the Dam1 complex forms a 16-unit ring encircling and tracking the tip of a MT in vitro, consistent with its cellular function as a coupler. Dam1 also forms smaller, nonring patches in vitro that track the dynamic ends of MTs. However, the identity of Dam1’s functional form in vivo remains unknown. Here we report a comprehensive in vivo characterization of Dam1 in the fission yeast Schizosaccharomyces pombe . In addition to their dense localizations on kinetochores and spindle MTs during mitosis, we identify that Dam1 is also localized onto cytoplasmic MTs as discrete spots in interphase, providing the unique opportunity to analyze Dam1 oligomers at the single-particle resolution in live cells. Such analysis shows that each oligomer contains one to five copies of Dam1, and is able to “switch-rail” while moving along MTs, precluding the possibility of a 16-unit encircling structure. Dam1 patches track the plus ends of the shortening, but not the elongating, MTs and retard MT depolymerization. Together with Mal3, the EB1-like MT-interacting protein, cytoplasmic Dam1 plays an important role in maintaining proper cell shape. In mitosis, kinetochore-associated Dam1 appears to facilitate kMT depolymerization. Together, our findings suggest that patches, instead of rings, are the physiologically functional forms of Dam1 in pombe. Our findings help establish the benchmark parameters of the Dam1 coupler and elucidate the mechanism of its functions.

show abstract

Microtubule Nucleation at Non-Spindle Pole Body Microtubule-Organizing Centers Requires Fission Yeast Centrosomin-Related Protein mod20p

Cited by 173 publications

References 58 publications

Ppc89 Links Multiple Proteins, Including the Septation Initiation Network, to the Core of the Fission Yeast Spindle-Pole Body

Ppc89 Links Multiple Proteins, Including the Septation Initiation Network, to the Core of the Fission Yeast Spindle-Pole Body

Generation of noncentrosomal microtubule arrays

A non-ring-like form of the Dam1 complex modulates microtubule dynamics in fission yeast

Contact Info

Product

Resources

About