A 47-kDa human nuclear protein recognized by antikinetochore autoimmune sera is homologous with the protein encoded by RCC1, a gene implicated in onset of chromosome condensation.

Assembly of the mitotic spindle requires a global change in the activity and constitution of the microtubule-binding-protein array at mitotic onset. An important subset of mitotic microtubule-binding proteins localises to the nucleus in interphase and essentially contributes to spindle formation and function after nuclear envelope breakdown. Here, we used a proteomic approach to selectively identify proteins of this category and revealed 50 poorly characterised human gene products, among them the echinoderm microtubule-associated-protein-like gene product, EML3. Indirect immunofluorescence showed that EML3 colocalises with spindle microtubules throughout all mitotic stages. In interphase, EML3 colocalised with cytoplasmic microtubules and accumulated in interphase nuclei. Using YFP-fusion constructs of EML3, we located a nuclear localisation signal and confirmed the microtubule-binding domain of EML3. Functional analysis of EML3 using time-lapse fluorescence microscopy and detailed end-point analysis of phenotypes after siRNA knockdown demonstrates an important role for EML3 in correct metaphase chromosome alignment. Our proteomic identification screen combined with sensitive phenotypic analysis therefore provides a reliable platform for the identification and characterisation of proteins important for correct cell division

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“…1B). The majority of RCC1, a chromatin-associated nuclear protein (Bischoff et al, 1990) was found in nuclear extracts (Fig. 1B).…”

Section: Resultsmentioning

confidence: 99%

EML3 is a nuclear microtubule-binding protein required for the correct alignment of chromosomes in metaphase

Tegha-Dunghu

Neumann

Reber

et al. 2008

Journal of Cell Science

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“…Alternatively, the entire Ran cycle might run in the vicinity of kinetochores. A long-neglected study showed convincingly that RCC1, the Ran GEF, is located at centromeres (Bischoff et al, 1990), where it was termed CENP-D (Kingwell and Rattner, 1987). At kinetochores, this cycle could be part of an uncharacterized switching mechanism that is required for kinetochore stability, microtubule nucleation, or modulation of the dynamic instability of kinetochore-associated microtubules.…”

Section: The Role Of Ran In Kinetochore Assembly and Functionmentioning

confidence: 99%

The dynamic kinetochore-microtubule interface

Maiato

DeLuca

Salmon

et al. 2004

Journal of Cell Science

357

379

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The kinetochore is a control module that both powers and regulates chromosome segregation in mitosis and meiosis. The kinetochore-microtubule interface is remarkably fluid, with the microtubules growing and shrinking at their point of attachment to the kinetochore. Furthermore, the kinetochore itself is highly dynamic, its makeup changing as cells enter mitosis and as it encounters microtubules. Active kinetochores have yet to be isolated or reconstituted, and so the structure remains enigmatic. Nonetheless, recent advances in genetic, bioinformatic and imaging technology mean we are now beginning to understand how kinetochores assemble, bind to microtubules and release them when the connections made are inappropriate, and also how they influence microtubule behaviour. Recent work has begun to elucidate a pathway of kinetochore assembly in animal cells; the work has revealed that many kinetochore components are highly dynamic and that some cycle between kinetochores and spindle poles along microtubules. Further studies of the kinetochore-microtubule interface are illuminating: (1) the role of the Ndc80 complex and components of the Ran-GTPase system in microtubule attachment, force generation and microtubule-dependent inactivation of kinetochore spindle checkpoint activity; (2) the role of chromosomal passenger proteins in the correction of kinetochore attachment errors; and (3) the function of microtubule plus-end tracking proteins, motor depolymerases and other proteins in kinetochore movement on microtubules and movement coupled to microtubule poleward flux.

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“…6. The DJ628 insertion is 54 bp upstream of the transcription start site of the CG11133 gene, which encodes a protein that shares a domain with RCC1, a regulator of chromatin condensation (Bischoff et al ., 1990). The DJ634 insertion is in the first intron of the LK6 gene (CG17342), which encodes the centrosomal microtubuleassociated LK6 serine /threonine protein kinase (Kidd & Raff, 1997).…”

Section: Molecular Analysis Of Insertion Sitesmentioning

confidence: 99%

Spatio‐temporal analysis of gene expression during aging in Drosophila melanogaster

et al. 2002

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SummaryThe relationship between gene expression and the regulation of longevity is poorly understood. Previous studies focusing on microarray or tissue-specific changes in gene expression as a function of age have provided evidence that gene expression is a dynamic process which is regulated, even late in an organism's lifespan. Using the enhancer-trap technique, a systematic analysis of the spatio-temporal regulation of gene expression in tissues of adult Drosophila is presented. As many as 80% of enhancer traps analysed displayed (some form of) transcriptional change with age. In some cases the rate of change in expression was found to correlate with changes in longevity under various conditions, suggesting that they may be indicators of 'physiological age' and therefore valuable markers for dissecting the aging process. Molecular analysis of enhancer traps that showed increased activity with age was performed to identify candidate genes that may be important in the regulation of longevity; we identified changes in reporters associated with immunity, microtubule organization and muscle function.

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A 47-kDa human nuclear protein recognized by antikinetochore autoimmune sera is homologous with the protein encoded by RCC1, a gene implicated in onset of chromosome condensation.

Cited by 88 publications

References 25 publications

EML3 is a nuclear microtubule-binding protein required for the correct alignment of chromosomes in metaphase

EML3 is a nuclear microtubule-binding protein required for the correct alignment of chromosomes in metaphase

The dynamic kinetochore-microtubule interface

Spatio‐temporal analysis of gene expression during aging in Drosophila melanogaster

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