The ATP‐dependent helicase RUVBL1/TIP49a associates with tubulin during mitosis

Gärtner, Wolfgang; Rossbacher, Joerg; Zierhut, Barbara; Daneva, Teodora; Base, Wolfgang; Weissel, M; Waldhäusl, W.; Pasternack, Mark S.; Wagner, Ludwig

doi:10.1002/cm.10136

Cited by 66 publications

(83 citation statements)

References 43 publications

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“…Taken together, these data suggest a new role for cohesin following sister chromatid cohesion establishment. Interestingly, another ATP-dependent DNA helicase, RUVBL1/TIP49a, was reported to be concentrated at the centrosomes and associate with mitotic spindles during early stages of mitosis (Gartner et al, 2003). RUVBL1 is essential for cellular division and proliferation (Lim et al, 2000), and it is suggested that this protein plays a role in microtubule organization during mitosis since it directly associates with ␣-and ␥-tubulin and was shown to increase tubulin polymerization when over-expressed in the presence of Paclitaxel, a microtubule stabilizer (Gartner et al, 2003).…”

Section: Discussionmentioning

confidence: 99%

The DNA helicase ChlR1 is required for sister chromatid cohesion in mammalian cells

Parish

Rosa

Wang

et al. 2006

Journal of Cell Science

100

114

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It has recently been suggested that the Saccharomyces cerevisiae protein Chl1p plays a role in cohesion establishment. Here, we show that the human ATPdependent DNA helicase ChlR1 is required for sister chromatid cohesion in mammalian cells. Localization studies show that ChlR1 diffusely coats mitotic chromatin in prophase and then translocates from the chromatids to concentrate at the spindle poles during the transition to metaphase. Depletion of ChlR1 protein by RNA interference results in mitotic failure with replicated chromosomes failing to segregate after a pro-metaphase arrest. We show that depletion also results in abnormal sister chromatid cohesion, determined by increased separation of chromatid pairs at the centromere. Furthermore, biochemical studies show that ChlR1 is in complex with cohesin factors Scc1, Smc1 and Smc3. We conclude that human ChlR1 is required for sister chromatid cohesion and, hence, normal mitotic progression. These functions are important to maintain genetic fidelity.

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Section: Discussionmentioning

confidence: 99%

The DNA helicase ChlR1 is required for sister chromatid cohesion in mammalian cells

Parish

Rosa

Wang

et al. 2006

Journal of Cell Science

100

114

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“…The repertoire of RuvBL1 functions was furthermore extended by observations that RuvBL1 is associated with the mitotic spindle and the centrosomes via interactions with tubulin (19). These results suggest that RuvBL1 may play an important role in mitosis.…”

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confidence: 82%

Crystal Structure of the Human AAA+ Protein RuvBL1

Matías

Gorynia

Donner³

et al. 2006

Journal of Biological Chemistry

154

288

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RuvBL1 is an evolutionarily highly conserved eukaryotic protein belonging to the AAA ؉ -family of ATPases (ATPase associated with diverse cellular activities). It plays important roles in essential signaling pathways such as the c-Myc and Wnt pathways in chromatin remodeling, transcriptional and developmental regulation, and DNA repair and apoptosis. Herein we present the three-dimensional structure of the selenomethionine variant of human RuvBL1 refined using diffraction data to 2.2 Å of resolution. The crystal structure of the hexamer is formed of ADP-bound RuvBL1 monomers. The monomers contain three domains, of which the first and the third are involved in ATP binding and hydrolysis. Although it has been shown that ATPase activity of RuvBL1 is needed for several in vivo functions, we could only detect a marginal activity with the purified protein. Structural homology and DNA binding studies demonstrate that the second domain, which is unique among AAA ؉ proteins and not present in the bacterial homolog RuvB, is a novel DNA/RNA-binding domain. We were able to demonstrate that RuvBL1 interacted with single-stranded DNA/RNA and double-stranded DNA. The structure of the RuvBL1⅐ADP complex, combined with our biochemical results, suggest that although RuvBL1 has all the structural characteristics of a molecular motor, even of an ATP-driven helicase, one or more as yet undetermined cofactors are needed for its enzymatic activity.RuvBL1 is a ubiquitously expressed protein (1) that plays important roles in chromatin remodeling, transcription, DNA repair, and apoptosis (2, 3). The significant evolutionary conservation of RuvBL1 from yeast to man strongly suggests that it mediates important cellular functions. RuvBL1 was originally identified by several unrelated approaches and is also known as TIP49a 3 (TATA-binding protein-interacting protein) (1, 4), Rvb1p (5), TAP54␣ (TIP60-associated protein) (3), and Pontin52 (1). The RuvBL1 gene is essential for viability in the yeast Saccharomyces cerevisiae (6), Drosophila melanogaster (7), and in Caenorhabditis elegans.4 A number of chromatin-remodeling complexes contain RuvBL1, like INO80 in yeast and human (8), which is involved in transcription and DNA repair (2), and p400 in animal cells (9). It was demonstrated that yeast Rvb1p is required for the catalytic activity of the INO80 chromatin-remodeling complex (8). RuvBL1 is also an essential component of the human histone acetylase/chromatin-remodeling complex TIP60 (3), which consists of at least 14 distinct subunits and displays histone acetylase activity on chromatin, ATPase, DNA helicase, and structural DNA binding activities.Generally, chromatin-remodeling complexes regulate chromatin structure and are critical for DNA-based transactions in the cell (10, 11). Acetylation of nucleosomal histones leads to relaxation of chromatin structure, thus enabling various transcription factors to gain access to chromatin and to interact with DNA (12, 13). As part of a chromatin-remodeling complex, yeast Rvb1p regulates the transcri...

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“…Immunofluorescence Microscopy-Immunofluorescence microscopy was performed as described previously (21). After deparaffinization, sections were incubated with a mixture of CLU H-330 rabbit polyclonal antibody (1:50 dilution) and monoclonal mouse anti-human CgA antibody (1:10 dilution).…”

Section: Methodsmentioning

confidence: 99%

Clusterin Expression in Normal Mucosa and Colorectal Cancer

Andersen

Schepeler

Thorsen

et al. 2007

Molecular & Cellular Proteomics

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The gene Clusterin is a target for cancer therapy in clinical trials. The indication for intervention is up-regulated Clusterin expression. Clusterin has been reported to be deregulated in multiple cancer types, including colorectal cancer (CRC). However, for CRC the studies have disagreed on whether Clusterin is up-or down-regulated by neoplastic cells. In the present study we sought to clarify the expression and distribution of Clusterin mRNAs and proteins in normal and neoplastic colorectal tissue through laser microdissection, variant-specific real time RT-PCR, immunohistochemistry, immunofluorescence, Western blotting, and array-based transcriptional profiling. At the transcript level we demonstrated the expression of two novel Clusterin transcripts in addition to the known transcript, and at the protein level we demonstrated two Clusterin isoforms. Our analysis of normal epithelial cells revealed that among these, Clusterin was only expressed by rare neuroendocrine subtype. Furthermore our analysis showed that in the normal mucosa the majority of the observed Clusterin protein originated from the stromal compartment. In tumors we found that Clusterin was de novo synthesized by non-neuroendocrine cancer cells in ϳ25% of cases. Moreover we found that the overall Clusterin level in tumors often appeared to be lower than in normal mucosa due to the stromal compartment often being suppressed in tumors. Although Clusterin in normal neuroendocrine cells showed a basal localization, the localization in cancer cells was often apical and in some cases associated with apical secretion. Collectively our results indicate that Clusterin expression is very complex.

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The ATP‐dependent helicase RUVBL1/TIP49a associates with tubulin during mitosis

Cited by 66 publications

References 43 publications

The DNA helicase ChlR1 is required for sister chromatid cohesion in mammalian cells

The DNA helicase ChlR1 is required for sister chromatid cohesion in mammalian cells

Crystal Structure of the Human AAA+ Protein RuvBL1

Clusterin Expression in Normal Mucosa and Colorectal Cancer

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