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Abstract LB-124: RHAMM acts near chromosomes to promote mitotic spindle assembly by regulating the location and abundance of TPX2 and the downstream activation of Aurora A.

2013· article· en· W2019200981 on OpenAlexaff
Helen Chen, Chinten James Lim, Christopher A. Maxwell

Bibliographic record

VenueCancer Research · 2013
Typearticle
Languageen
FieldBiochemistry, Genetics and Molecular Biology
TopicMicrotubule and mitosis dynamics
Canadian institutionsChild and Family Research Institute
Fundersnot available
KeywordsCentrosomeCell biologyKinetochoreMitosisSpindle apparatusSpindle checkpointSpindle pole bodyMultipolar spindlesBiologyAurora A kinaseAurora B kinaseMicrotubuleCell cycleCell divisionCellGeneticsGene

Abstract

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Abstract Enhanced cell proliferation is a hallmark of cancer. Thus, molecular pathways underlying mitosis are important targets for therapy. To form a bipolar mitotic spindle, the dividing cell must assemble microtubules at centrosomes [spindle poles] as well as non-centrosome sites [kinetochores]. The spindle then captures and aligns the duplicated chromosomes. Correct attachment satisfies the spindle assembly checkpoint (SAC) and allows for mitotic progression. Aurora kinase A, when bound by its activator, TPX2, is a key regulator of spindle assembly. Upon activation, the kinase phosphorylates substrates to target them to the centrosomes, and protects them from proteolytic degradation. Since it is necessary for optimal Aurora A function, TPX2 availability is tightly controlled. The receptor for hyaluronan-mediated motility (RHAMM) is a microtubule associated protein that interacts with TPX2 and participates in spindle assembly. However, it is unclear whether or how the RHAMM-TPX2 complex modulates Aurora A activity. We hypothesize that RHAMM interacts with TPX2 to determine TPX2 location and abundance, which facilitates Aurora A activity and spindle assembly. We have followed the location and interactions of RHAMM through mitosis, as well as the consequences for its loss of function. We tracked the location of RHAMM and GFP-RHAMM and found it at the kinetochores. Then, we colocalized RHAMM with key kinetochore proteins, BUBR1 and NDC80, during microtubule nucleation and confirmed physical associations by IP. For insights into the function of RHAMM, we followed mitotic cells in real-time. In HeLa cells treated with siRNA targeting RHAMM, we found a significant increase in aberrant spindle morphology and mitotic failure, and we were able to rescue these phenotypes by reintroducing GFP-RHAMM. We also measured the duration of spindle assembly, chromosome congression and cytokinesis. RHAMM depleted mitotic cells present two distinct phenotypes, and these cells either fail spindle assembly or construct phenotypically “normal” mitotic spindles that do not satisfy the SAC. Thus, the protein's location, interactions, and the consequences of its depletion all point to a critical role at the kinetochore. Previous reports suggest RHAMM participates in spindle assembly by interacting with TPX2, however, the exact mechanism remains unknown. We find that RHAMM depletion decreases TPX2 protein abundance during early mitosis, and impairs TPX2 transit to the spindle poles, which results in reduced Aurora A protein abundance and activity. Together, these results suggest that RHAMM influences Aurora A actions by regulating the stability and location of TPX2. Further insights into RHAMM associated pathways may lead to the discovery of new therapeutic targets and optimize the usage of promising drugs such as Aurora A inhibitors. Citation Format: Helen Chen, Chinten Lim, Chris Maxwell. RHAMM acts near chromosomes to promote mitotic spindle assembly by regulating the location and abundance of TPX2 and the downstream activation of Aurora A. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr LB-124. doi:10.1158/1538-7445.AM2013-LB-124

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How this classification was reachedexpand

Full frame machine prediction

Teacher imitation

Not calibrated prevalence, not ground truth. Human validation pending. The Gemma side is a direct model label for every work in the frame, read from the title-only record. The Codex side is a classifier learned from the 10,348 direct Codex labels and calibrated to design-weighted sample rates; fields without enough sample support carry no Codex call. Candidate is the union of the two sides; consensus is their intersection. These outputs are machine_predicted_unvalidated and are not human labels.

metaresearch head score (Codex)0.000
metaresearch head score (Gemma)0.000
Version: metacan-v3-hybrid-931329e0061cValidation status: machine_predicted_unvalidated
Candidate categoriesnone
Consensus categoriesnone
DomainCandidate signal: none · Consensus signal: none
Study designCandidate signal: Bench or experimental · Consensus signal: Bench or experimental
GenreCandidate signal: Empirical · Consensus signal: Empirical
Teacher disagreement score0.005
Threshold uncertainty score0.016

Distilled classifier scores by category (both heads)

CategoryCodexGemma
Metaresearch0.0000.000
Meta-epidemiology (narrow)0.0000.000
Meta-epidemiology (broad)0.0000.000
Bibliometrics0.0000.000
Science and technology studies0.0000.000
Scholarly communication0.0000.000
Open science0.0000.000
Research integrity0.0000.001
Insufficient payload (model declined to judge)0.0050.001

Machine scores (provisional)

The two teacher heads of the student model, read on this work. A score orders the frame for review; it never asserts a category, and the validation status ships verbatim with every row.

Baseline scores from an immature model (maturity gate not passed, 7 training rounds). Scores rank; they never assert a category.

Opus teacher head0.018
GPT teacher head0.314
Teacher spread0.296 · how far apart the two teachers sit on this one work
Validation statusscore_only:v0-immature-baseline · verbatim from the scoring run: score_only means the number may rank works, and no category label ships from it

Classification

machine, unvalidated

Machine predicted; a candidate call from one source (direct Gemma or distilled Codex), not a consensus.

The models applied no category: nothing in the taxonomy fit this work.
Study designBench or experimental
Domainnot available
GenreEmpirical

How this classification was reached, model by model and score by score, is at the end of the page under "How this classification was reached".

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Citations0
Published2013
Admission routes1
Has abstractyes

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