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PMID:21852952

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Citation

Hughes, SE, Beeler, JS, Seat, A, Slaughter, BD, Unruh, JR, Bauerly, E, Matthies, HJ and Hawley, RS (2011) Gamma-tubulin is required for bipolar spindle assembly and for proper kinetochore microtubule attachments during prometaphase I in Drosophila oocytes. PLoS Genet. 7:e1002209

Abstract

In many animal species the meiosis I spindle in oocytes is anastral and lacks centrosomes. Previous studies of Drosophila oocytes failed to detect the native form of the germline-specific γ-tubulin (γTub37C) in meiosis I spindles, and genetic studies have yielded conflicting data regarding the role of γTub37C in the formation of bipolar spindles at meiosis I. Our examination of living and fixed oocytes carrying either a null allele or strong missense mutation in the γtub37C gene demonstrates a role for γTub37C in the positioning of the oocyte nucleus during late prophase, as well as in the formation and maintenance of bipolar spindles in Drosophila oocytes. Prometaphase I spindles in γtub37C mutant oocytes showed wide, non-tapered spindle poles and disrupted positioning. Additionally, chromosomes failed to align properly on the spindle and showed morphological defects. The kinetochores failed to properly co-orient and often lacked proper attachments to the microtubule bundles, suggesting that γTub37C is required to stabilize kinetochore microtubule attachments in anastral spindles. Although spindle bipolarity was sometimes achieved by metaphase I in both γtub37C mutants, the resulting chromosome masses displayed highly disrupted chromosome alignment. Therefore, our data conclusively demonstrate a role for γTub37C in both the formation of the anastral meiosis I spindle and in the proper attachment of kinetochore microtubules. Finally, multispectral imaging demonstrates the presences of native γTub37C along the length of wild-type meiosis I spindles.

Links

PubMed PMC3154956 Online version:10.1371/journal.pgen.1002209

Keywords

Animals; Cell Cycle Checkpoints; Chromosomes/metabolism; Drosophila Proteins/metabolism; Drosophila melanogaster/cytology; Drosophila melanogaster/genetics; Drosophila melanogaster/physiology; Female; Kinetochores/metabolism; Male; Meiosis; Metaphase; Microtubules/metabolism; Mitotic Spindle Apparatus/metabolism; Mutation, Missense; Oocytes/metabolism; Oocytes/physiology; Prometaphase; Protein Binding; Tubulin/genetics; Tubulin/metabolism

Significance

Annotations

Gene product Qualifier GO Term Evidence Code with/from Aspect Extension Notes Status

DROME:TBG2

located_in

GO:0072687: meiotic spindle

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

DROME:TBG2

involved_in

GO:0007312: oocyte nucleus migration involved in oocyte dorsal/ventral axis specification

ECO:0000315: mutant phenotype evidence used in manual assertion

P

Seeded From UniProt

complete

DROME:TBG2

involved_in

GO:0007057: spindle assembly involved in female meiosis I

ECO:0000315: mutant phenotype evidence used in manual assertion

P

Seeded From UniProt

complete

DROME:TBG2

involved_in

GO:0051316: attachment of spindle microtubules to kinetochore involved in meiotic chromosome segregation

ECO:0000315: mutant phenotype evidence used in manual assertion

P

Seeded From UniProt

complete


See also

References

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