A novel magnetic resonance-based method to measure gene expression in living cells

In unicellular and multicellular eukaryotes, elaborate gene regulatory mechanisms facilitate a broad range of biological processes from cell division to morphological differentiation. In order to fully understand the gene regulatory networks involved in these biological processes, the spatial and temporal patterns of expression of many thousands of genes will need to be determined in real time in living organisms. Currently available techniques are not sufficient to achieve this goal; however, novel methods based on magnetic resonance (MR) imaging may be particularly useful for sensitive detection of gene expression in opaque tissues. This report describes a novel reporter gene system that monitors gene expression dynamically and quantitatively, in yeast cells, by measuring the accumulation of inorganic polyphosphate (polyP) using MR spectroscopy (MRS) or MR spectroscopic imaging (MRI). Because this system is completely non-invasive and does not require exogenous substrates, it is a powerful tool for studying gene expression in multicellular organisms, as well

A pre-metazoan origin of the CRK gene family and co-opted signaling network.

CRK and CRKL adapter proteins play essential roles in development and cancer through their SRC homology 2 and 3 (SH2 and SH3) domains. To gain insight into the origin of their shared functions, we have investigated their evolutionary history. We propose a term, crk/crkl ancestral (crka), for orthologs in invertebrates before the divergence of CRK and CRKL in the vertebrate ancestor. We have isolated two orthologs expressed in the choanoflagellate Monosiga brevicollis, a unicellular relative to the metazoans. Consistent with its highly-conserved three-dimensional structure, the SH2 domain of M. brevicollis crka1 can bind to the mammalian CRK/CRKL SH2 binding consensus phospho-YxxP, and to the SRC substrate/focal adhesion protein BCAR1 (p130(CAS)) in the presence of activated SRC. These results demonstrate an ancient origin of the CRK/CRKL SH2-target recognition specificity. Although BCAR1 orthologs exist only in metazoans as identified by an N-terminal SH3 domain, YxxP motifs, and a C-terminal FAT-like domain, some pre-metazoan transmembrane proteins include several YxxP repeats in their cytosolic region, suggesting that they are remotely related to the BCAR1 substrate domain. Since the tyrosine kinase SRC also has a pre-metazoan origin, co-option of BCAR1-related sequences may have rewired the crka-dependent network to mediate adhesion signals in the metazoan ancestor

Essential Role of the \u3ci\u3eCrk\u3c/i\u3e Family-Dosage in DiGeorge-Like Anomaly and Metabolic Homeostasis

CRK and CRKL (CRK-like) encode adapter proteins with similar biochemical properties. Here, we show that a 50% reduction of the family-combined dosage generates developmental defects, including aspects of DiGeorge/del22q11 syndrome in mice. Like the mouse homologs of two 22q11.21 genes CRKL and TBX1, Crk and Tbx1 also genetically interact, thus suggesting that pathways shared by the three genes participate in organogenesis affected in the syndrome. We also show that Crk and Crkl are required during mesoderm development, and Crk/Crkl deficiency results in small cell size and abnormal mesenchyme behavior in primary embryonic fibroblasts. Our systems-wide analyses reveal impaired glycolysis, associated with low Hif1a protein levels as well as reduced histone H3K27 acetylation in several key glycolysis genes. Furthermore, Crk/Crkl deficiency sensitizes MEFs to 2deoxy-D-glucose, a competitive inhibitor of glycolysis, to induce cell blebbing. Activated Rapgef1, a Crk/Crkl-downstream effector, rescues several aspects of the cell phenotype, including proliferation, cell size, focal adhesions, and phosphorylation of p70 S6k1 and ribosomal protein S6. Our investigations demonstrate that Crk/Crkl-shared pathways orchestrate metabolic homeostasis and cell behavior through widespread epigenetic controls

Perceived managerial and leadership effectiveness in a Korean context: An indigenous qualitative study

Multinational corporations (MNCs) across the world have sent an increasing number of managers abroad to leverage unprecedented opportunities in the era of globalization. However, their failure rate has been above 33% for decades, resulting in substantial costs (Puck, Kittler, & Wright, 2008). One of the primary reasons for this failure is a lack of understanding of the national and organizational cultures within the host countries (Festing & Maletzky, 2011). For example, while a number of MNCs have entered the Korean market, several such as Yahoo, Motorola, and Walmart have failed and withdrawn due to the companies’ lack of adjustment to the Korean cultural context (Choe, 2006; Woo, 2013). In spite of the significance of culturally embedded practices, most researchers who have explored management and leadership in Asian countries, whether they were Western or indigenous researchers, have implemented studies using extant Western management and leadership theories derived within the Western cultural context (Leung, 2007; Tsui, 2006). Numerous scholars have claimed that this could be problematic because the findings of such studies may not be applicable to non-Western countries (Li, 2012; Liden & Antonakis, 2009), and may fail to provide insights and understanding of novel contexts or to reveal indigenous aspects of management and leadership (Tsui, 2007). Consequently, there have been increasing calls for indigenous management and leadership research within Asian countries (see Li et al., 2014; Lyles, 2009; Tsui, 2004; Wolfgramm, Spiller, & Voyageur, 2014). Over the past 30 years, managerial effectiveness and leadership effectiveness have been substantially neglected areas of management research (Noordegraaf & Stewart, 2000; Yukl, Gordon, & Taber, 2002). In addition, there has been little agreement on what specific behaviors distinguish effective managers from ineffective ones. Furthermore, more research is needed to examine the managerial and leadership behaviors that are critical for shaping the performance of individuals, groups and organizations (see Borman & Brush, 1993; Cammock, Nilakant & Dakin, 1995; Mumford, 2011; Noordegraaf & Stewart, 2000; Yukl et al., 2002). While most of the research related to managerial and leadership effectiveness has been conducted in the U.S., the few notable non-U.S. studies include that of Cammock et al. (1995) in New Zealand who developed a behavioral lay model of managerial effectiveness using the repertory grid technique. Another notable exception is the cumulative series of perceived managerial and leadership effectiveness studies conducted by Hamlin with various indigenous co-researchers in Western and non-Western countries (see Hamlin & Patel, 2012; Ruiz, Wang, & Hamlin, 2013) using Flanagan’s (1954) critical incident technique (CIT)

Tunable Superhydrophobic and Optical Properties of Colloidal Films Coated with Block Copolymer Micelle/Micelle Multilayers

Superhydrophobic surfaces are prepared based on silica colloids coated with multiple layers of block copolymer micelles (see figure). The hierarchical surface roughness can be tuned by adjusting the pH and molecular weight of the polymer segments. Optical functionality is imparted to the films by the inclusion of quantum dots or organic dyes in the hydrophobic cores, as depicted in the figure.This work was supported by a Korea Science and Engineering Foundation (KOSEF) grant funded by the Ministry of Science and Technology (R17-2007-059-01 000-0). This work was also supported by the Brain Korea 21 Program endorsed by the Ministry of Education of Korea, the Seoul Science Fellowship by the Seoul Metropolitan Government, ERC (CMPS, Center for Materials and Processes of Self-Assembly) Program of the MOST/KOSEF (R11-2005-048-00000-0), the New Faculty Research Program 2006 of Kookmin University in Korea, and the Australian Research Council. Additional support for this work was provided by a Korea Research Foundation Grant funded by the Korean Government (KRF-2006-311-D00453)

Assembly of Regulatory Factors on rRNA and Ribosomal Protein Genes in Saccharomyces cerevisiae▿ †

HMO1 is a high-mobility group B protein that plays a role in transcription of genes encoding rRNA and ribosomal proteins (RPGs) in Saccharomyces cerevisiae. This study uses genome-wide chromatin immunoprecipitation to study the roles of HMO1, FHL1, and RAP1 in transcription of these genes as well as other RNA polymerase II-transcribed genes in yeast. The results show that HMO1 associates with the 35S rRNA gene in an RNA polymerase I-dependent manner and that RPG promoters (138 in total) can be classified into several distinct groups based on HMO1 abundance at the promoter and the HMO1 dependence of FHL1 and/or RAP1 binding to the promoter. FHL1, a key regulator of RPGs, binds to most of the HMO1-enriched and transcriptionally HMO1-dependent RPG promoters in an HMO1-dependent manner, whereas it binds to HMO1-limited RPG promoters in an HMO1-independent manner, irrespective of whether they are transcribed in an HMO1-dependent manner. Reporter gene assays indicate that these functional properties are determined by the promoter sequence

measure gene expression

novel magnetic resonance-based method t

Regulation of B cell activating factor (BAFF) receptor expression by NF-κB signaling in rheumatoid arthritis B cells

B cells play an important role in the pathogenesis of rheumatoid arthritis (RA). High levels of B cell activating factor (BAFF) are detected in autoimmune diseases. BAFF and BAFF receptor (BAFF-R) are expressed in B and T cells of RA synovium. The study was undertaken to identify the NF-κB signal pathway involved in the induction of BAFF-R in human B cells. Immunohistochemical staining of NF-κB p65, NF-κB p50, BAFF, and BAFF-R was performed on sections of synovium from severe and mild RA and osteoarthritis (OA) patients. Peripheral blood mononuclear cells (PBMCs) were isolated from control and RA patients and B cells were isolated from controls. BAFF-R was analyzed by flow cytometry, realtime PCR and confocal staining after treatment with NF-κB inhibitors. NF-κB p65, NF-κB p50, BAFF, and BAFF-R were highly expressed in severe RA synovium relative to mild RA synovium or OA synovium. BAFF-R expression was reduced by NF-κB inhibitors in PBMCs and B cells from normal controls. We also showed reduction in expression of BAFF-R via inhibition of the NF-κB pathway in PBMCs of RA patients. BAFF/BAFF-R signaling is an important mechanism of pathogenesis in RA and that BAFF-R reduction by NF-κB blocking therapy is another choice for controlling B cells in autoimmune diseases such as RA