Asymmetric mitotic segregation of the yeast spindle pole body

Vallen, Elizabeth A.; Scherson, Talma; Roberts, Tom; Zee, Karen van; Rose, Mark D.

doi:10.1016/0092-8674(92)90451-h

Cited by 92 publications

(70 citation statements)

References 28 publications

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“…We have never observed two or more dots in a cell (n > 1,000). In this regard the Jnmlp:/3-galactosidase protein most closely resembles a Karl-/3-galactosidase fusion protein that localizes asymmetrically to the single SPB nearest the bud (Vallen et al, 1992). The localization of the Jnmlp:~-galactosidase fusion is clearly different in that it is not always associated with a SPB.…”

Section: A Jnmlp:fl-galactosidase Fusion Protein Has a Restricted Dismentioning

confidence: 87%

The JNM1 gene in the yeast Saccharomyces cerevisiae is required for nuclear migration and spindle orientation during the mitotic cell cycle.

McMillan

Tatchell

1994

The Journal of Cell Biology

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Abstract. JNM1, a novel gene on chromosome XIII in the yeast Saccharomyces cerevisiae, is required for proper nuclear migration, jnml null mutants have a temperature-dependent defect in nuclear migration and an accompanying alteration in astral microtubules. At 30°C, a significant proportion of the mitotic spindles is not properly located at the neck between the mother cell and the bud. This defect is more severe at low temperature. At ll°C, 60% of the cells accumulate with large buds, most of which have two DAPI staining regions in the mother cell. Although mitosis is delayed and nuclear migration is defective in jnml mutants, we rarely observe more than two nuclei in a cell, nor do we frequently observe anuclear cells. No loss of viability is observed at ll°C and cells continue to grow exponentially with increased doubling time. T HE nuclei in the budding yeast Saccharomyces cerevisiae migrate to the neck between the mother cell and the bud prior to mitosis and the spindle microtubules orient along the long axis of the cell. At mitosis the spindle elongates through the neck between the mother cell and the bud and chromosomes separate, depositing one set of chromosomes in the mother cell body and one set in the bud. A key role for microtubules in this process has been shown by the inhibitory effects on nuclear migration by microtubule inhibitors (Jacobs et al., 1988) and by defects in nuclear migration exhibited by some B tubulin mutants (Huffaker et al., 1988). Cytoplasmic (or astral) microtubules appear to be mainly responsible for nuclear migration.Mutant alleles of the tub2 gene that impart a specific defect in astral microtubule stability show defects in nuclear migration during mitosis or karyogamy, but have relatively little

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Section: A Jnmlp:fl-galactosidase Fusion Protein Has a Restricted Dismentioning

confidence: 87%

The JNM1 gene in the yeast Saccharomyces cerevisiae is required for nuclear migration and spindle orientation during the mitotic cell cycle.

McMillan

Tatchell

1994

The Journal of Cell Biology

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“…For example, it has not been possible to localize the authentic Karl or Kar3 proteins, whereas fusion proteins have been localized (Meluh and Rose 1990;Vallen et al 1992). In the case of Cikl, ~-gal fusions localize to microtubules, whereas the authentic protein targets to the spindle pole body (Page et al 1994).…”

Section: Advantages and Disadvantages Of ~-Gal Fusionsmentioning

confidence: 99%

“…comm. ), spindle pole body (Vallen et al 1992), microtubules (Meluh and Rose 1990;Page et al 1994), and sites of polarized growth (Trueheart et al 1987;C. Costigan and M. Snyder, unpubl.).…”

Section: Localization Of Fusion Proteinsmentioning

confidence: 99%

Large-scale analysis of gene expression, protein localization, and gene disruption in Saccharomyces cerevisiae.

Burns¹,

Grimwade²,

et al. 1994

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We have developed a large-scale screen to identify genes expressed at different times during the life cycle of Saccharomyces cerevisiae and to determine the subcellular locations of many of the encoded gene products. Diploid yeast strains containing random lacZ insertions throughout the genome have been constructed by transformation with a mutagenized genomic library. Twenty-eight hundred transformants containing fusion genes expressed during vegetative growth and 55 transformants containing meiotically induced fusion genes have been identified. Based on the frequency of transformed strains producing 13-galactosidase, we estimate that 80-86% of the yeast genome (excluding the rDNA) contains open reading frames expressed in vegetative cells and that there are 93-135 meiotically induced genes. Indirect immunofluorescence analysis of 2373 strains carrying fusion genes expressed in vegetative cells has identified 245 fusion proteins that localize to discrete locations in the cell, including the nucleus, mitochondria, endoplasmic reticulum, cytoplasmic dots, spindle pole body, and microtubules. The DNA sequence adjacent to the lacZ gene has been determined for 91 vegetative fusion genes whose products have been localized and for 43 meiotically induced fusions. Although most fusions represent genes unidentified previously, many correspond to known genes, including some whose expression has not been studied previously and whose products have not been localized. For example, Sec21-13-gal fusion proteins yield a Golgi-like staining pattern, Tyl-13-gal fusion proteins localize to cytoplasmic dots, and the meiosis-specific Mekl/Mre4-13-gal and Spol 1-13-gal fusion proteins reside in the nucleus. The phenotypes in haploid cells have been analyzed for 59 strains containing chromosomal fusion genes expressed during vegetative growth; 9 strains fail to form colonies indicating that the disrupted genes are essential. Fifteen additional strains display slow growth or are impaired for growth on specific media or in the presence of inhibitors. Of 39 meiotically induced fusion genes examined, 14 disruptions confer defects in spore formation or spore viability in homozygous diploids. Our results will allow researchers who identify a yeast gene to determine immediately whether that gene is expressed at a specific time during the life cycle and whether its gene product localizes to a specific subcellular location.

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“…Once DNA replication is completed, SPBs separate and a short intranuclear spindle forms. Spindle polarity is already evident during SPB separation (Vallen et al, 1992), and results in a distinct pattern of cytoplasmic microtubule-cortex interactions throughout assembly: either with the bud cortex (SPBdaughter ) or the bud neck region (SPB mother ), respectively (Segal et al, 2000). These dynamic interactions play a critical role in spindle orientation (Carminati and Stearns, 1997;Shaw et al, 1997).…”

Section: Introductionmentioning

confidence: 99%

Bud6 Directs Sequential Microtubule Interactions with the Bud Tip and Bud Neck during Spindle Morphogenesis inSaccharomyces cerevisiae

Segal

Bloom

Reed

2000

MBoC

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In budding yeast, spindle polarity relies on a precise temporal program of cytoplasmic microtubule-cortex interactions throughout spindle assembly. Loss of Clb5-dependent kinase activity under conditions of attenuated Cdc28 function disrupts this program, resulting in diploid-specific lethality. Here we show that polarity loss is tolerated by haploids due to a more prominent contribution of microtubule-neck interactions to spindle orientation inherent to haploids. These differences are mediated by the relative partition of Bud6 between the bud tip and bud neck, distinguishing haploids from diploids. Bud6 localizes initially to the bud tip and accumulates at the neck concomitant with spindle assembly. bud6⌬ mutant phenotypes are consistent with Bud6's role as a cortical cue for cytoplasmic microtubule capture. Moreover, mutations that affect Bud6 localization and partitioning disrupt the sequential program of microtubule-cortex interactions accordingly. These data support a model whereby Bud6 sequentially cues microtubule capture events at the bud tip followed by capture events at the bud neck, necessary for correct spindle morphogenesis and polarity.

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Asymmetric mitotic segregation of the yeast spindle pole body

Cited by 92 publications

References 28 publications

The JNM1 gene in the yeast Saccharomyces cerevisiae is required for nuclear migration and spindle orientation during the mitotic cell cycle.

The JNM1 gene in the yeast Saccharomyces cerevisiae is required for nuclear migration and spindle orientation during the mitotic cell cycle.

Large-scale analysis of gene expression, protein localization, and gene disruption in Saccharomyces cerevisiae.

Bud6 Directs Sequential Microtubule Interactions with the Bud Tip and Bud Neck during Spindle Morphogenesis inSaccharomyces cerevisiae

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