Li, F. et al. p21-activated kinase 1 interacts with and phosphorylates histone H3 in breast cancer cells. EMBO Rep. 3, 767-773

The University of Texas MD Anderson Cancer Center, Houston 77030, USA.
EMBO Reports (Impact Factor: 9.06). 09/2002; 3(8):767-73. DOI: 10.1093/embo-reports/kvf157
Source: PubMed


Stimulation of p21-activated kinase-1 (Pak1) signaling promotes motility, invasiveness, anchorage-independent growth and abnormal mitotic assembly in human breast cancer cells. Here, we provide new evidence that, before the onset of mitosis, activated Pak1 is specifically localized with the chromosomes during prophase and on the centrosomes in metaphase and moves to the contraction ring during cytokinesis. To identify mitosis-specific substrates of Pak1, we screened a synchronized G2-M expression library by using a glutathione transferase Pak1 solid-phase-based kinase reaction. This analysis identified histone H3 as a substrate of Pak1 both in vitro and in vivo, and it specifically interacted with Pak1 but not Pak2 or Pak3. Site-directed mutagenesis indicated that Pak1 phosphorylates histone H3 on Ser10. Expressions of the wild-type, or catalytically active, Pak1 caused it to appear at the poles corresponding to mitotic centrosomes in a variety of mammalian cells. Together, these results suggest for the first time that Pak1 interacts with and phosphorylates histone H3 and may thus influence the Pak1-histone H3 pathway, which in turn may influence mitotic events in breast cancer cells.

Download full-text


Available from: Jonathan Chernoff, Oct 01, 2015
31 Reads
    • "For example, PAK1 is located in the dendrites and axons of hippocampal and cortical neurons, while PAK3 accumulates in the cell bodies (Hayashi et al. 2002 ; Ong et al. 2002 ). Moreover, only PAK1 can shuttle to the nucleus and consequently regulate gene transcription (Li et al. 2002 ; Singh et al. 2005 ). PAKs bind to Rho family GTPases (RhoA, Rac1, and Cdc42) and they participate in a number of processes associated with cytoskeletal reorganization, including the regulation of cell proliferation and migration, neurogenesis , developmental apoptosis, neuronal polarity, neurite outgrowth, dendritic branching, spine formation, axonal guidance, and synaptic plasticity [reviewed by Kreis and Barnier ( 2009 )]. "
    [Show abstract] [Hide abstract]
    ABSTRACT: DSCAMs (Down syndrome cell adhesion molecules) are a group of immunoglobulin-like transmembrane proteins that contain fibronectin III domains. The founding member of the family was isolated in a positional cloning study that sought to identify genes located on chromosome 21 at the locus 21q22.2-q22.3 that is implicated in the neurological and cardiac phenotypes associated with Down's syndrome. In Drosophila, Dscam proteins are involved in neuronal wiring, while in vertebrates, the role of these cell adhesion molecules in neurogenesis, dendritogenesis, axonal outgrowth, synaptogenesis, and synaptic plasticity is only just beginning to be understood. In this chapter, we will review the functions ascribed to the two paralogous proteins found in humans, DSCAM and DSCAML1 (DSCAM-like 1), based on findings in knockout mice. The signaling pathways downstream of DSCAM activation and the role of DSCAM miss-expression in disease will be also discussed, particularly with regard to the intellectual disability in Down's syndrome.
    Cell Adhesion Molecules: Implications in Neurological Diseases, Edited by Vladimir Berezin and Peter S. Walmod, 10/2014: chapter 11: pages 249-270; Springer., ISBN: 978-1-4614-8089-1
  • Source
    • "Accumulating evidence has pointed out that there are several potential mechanisms responsible for PAK1 regulation of gene transcription. First, nuclear PAK1 associates with chromatin and regulates histone modifications such as phosphorylation of histone H3 at serine 10, which in turn directs gene transcription [13], [14]; second, PAK1 may interact with and phosphorylate transcription factors, transcription coregulators, or chromatin remodeling factors, thus regulating the bindings of the transcription machinery to nucleosomal DNA and consequently, gene transcription. A case in point is that PAK1 interacts with and phosphorylates transcriptional corepressor SHARP and enhances SHARP-mediated repression in Notch signaling [49]. "
    [Show abstract] [Hide abstract]
    ABSTRACT: P21-activated kinase 1 (PAK1), a serine/threonine protein kinase, modulates many cellular processes by phosphorylating its downstream substrates. In addition to its role in the cytoplasm, PAK1 also affects gene transcription due to its nuclear localization and association with chromatin. It is now recognized that PAK1 kinase activity and its nuclear translocation are rapidly stimulated by ionizing radiation (IR), and that PAK1 activation is a component of the DNA damage response. Owing to the role of PAK1 in the cell survival, its association with the chromatin, and now, stimulation by ionizing radiation, we hypothesize that PAK1 may be contributing to modulation of genes with roles in cellular processes that might be important in the DNA damage response. The purpose of this study was to identify new PAK1 targets in response to ionizing radiation with putative role in the DNA damage response. We examined the effect of IR on the gene expression patterns in the murine embryonic fibroblasts with or without Pak1 using microarray technology. Differentially expressed transcripts were identified using Gene Spring GX 10.0.2. Pathway, network, functional analyses and gene family classification were carried out using Kyoto Encyclopedia of Genes and Genomes (KEGG), Ingenuity Pathway, Gene Ontology and PANTHER respectively. Selective targets of PAK1 were validated by RT-qPCR. For the first time, we provide a genome-wide analysis of PAK1 and identify its targets with potential roles in the DNA damage response. Gene Ontology analysis identified genes in the IR-stimulated cells that were involved in cell cycle arrest and cell death. Pathway analysis revealed p53 pathway being most influenced by IR responsive, PAK1 targets. Gene family of transcription factors was over represented and gene networks involved in DNA replication, repair and cellular signaling were identified. In brief, this study identifies novel PAK1 dependent IR responsive genes which reveal new aspects of PAK1 biology.
    PLoS ONE 08/2013; 8(8):e66585. DOI:10.1371/journal.pone.0066585 · 3.23 Impact Factor
  • Source
    • "Image analysis showed that group I PAKs and their phosphorylated forms, hereafter named PAKs and PhoPAKs, respectively, localize in the cytoplasm as well as in the nucleus of HeLa cells (Figure 2A and E, respectively), as previously described [35-37]. PAKs and PhoPAKs cluster in spots of different dimensions in the nucleus (Figure 2A and E, respectively). "
    [Show abstract] [Hide abstract]
    ABSTRACT: Farnesyltransferase inhibitors (FTIs) are anticancer agents with a spectrum of activity in Ras-dependent and independent tumor cellular and xenograph models. How inhibition of protein farnesylation by FTIs results in reduced cancer cell proliferation is poorly understood due to the multiplicity of potential FTase targets. The low toxicity and oral availability of FTIs led to their introduction into clinical trials for the treatment of breast cancer, hematopoietic malignancy, advanced solid tumor and pancreatic cancer treatment, and Hutchinson-Gilford Progeria Syndrome. Although their efficacy in combinatorial therapies with conventional anticancer treatment for myeloid malignancy and solid tumors is promising, the overall results of clinical tests are far below expectations. Further exploitation of FTIs in the clinic will strongly rely on understanding how these drugs affect global cellular activity. Using FTase inhibitor I and genome-wide chemical profiling of the yeast barcoded deletion strain collection, we identified genes whose inactivation increases the antiproliferative action of this FTI peptidomimetic. The main findings were validated in a panel of cancer cell lines using FTI-277 in proliferation and biochemical assays paralleled by multiparametric image-based analyses. ABC transporter Pdr10 or p-21 activated kinase (PAK) gene deletion increases the antiproliferative action of FTase inhibitor I in yeast cells. Consistent with this, enhanced inhibition of cell proliferation by combining group I PAK inhibition, using IPA3, with FTI-277 was observed in melanoma (A375MM), lung (A549) and colon (HT29), but not in epithelial (HeLa) or breast (MCF7), cancer cell lines. Both HeLa and A375MM cells show changes in the nuclear localization of group 1 PAKs in response to FTI-277, but up-regulation of PAK protein levels is observed only in HeLa cells. Our data support the view that group I PAKs are part of a pro-survival pathway activated by FTI treatment, and group I PAK inactivation potentiates the anti-proliferative action of FTIs in yeast as well as in cancer cells. These findings open new perspectives for the use of FTIs in combinatorial strategies with PAK inhibitors in melanoma, lung and colon malignancy.
    Molecular Cancer 08/2013; 12(1):88. DOI:10.1186/1476-4598-12-88 · 4.26 Impact Factor
Show more