Cytoskeletal changes regulated by the PAK4 serine/threonine kinase are mediated by LIM kinase 1 and cofilin.
Authors Dan C; Kelly A; Bernard O; Minden A;
JournalJ Biol Chem
PubMed ID11413130
'PAK4 is the most recently identified member of the PAK family of serine/threonine kinases. PAK4 differs from other members of the PAK family in sequence and in many of its functions. Previously, we have shown that an important function of this kinase is to mediate the induction of filopodia in ... More
Sp3 Represses Gene Expression via the Titration of Promoter-specific Transcription Factors.
Authors Kennett Sarah B; Moorefield K Scott; Horowitz Jonathan M;
JournalJ Biol Chem
PubMed ID11773047
'We have determined previously that Sp3 encodes three distinct gene products as follows: a full-length protein (Sp3) that is an activator of transcription and two isoforms (M1 and M2) derived via internal translational initiation that function as transcriptional repressors. To identify amino acids and functions required for transcriptional repression, we ... More
Identification of phosphorylated residues that affect the activity of the mitotic kinase Aurora-A.
Authors Littlepage Laurie E; Wu Hua; Andresson Thorkell; Deanehan Julia K; Amundadottir Laufey T; Ruderman Joan V;
JournalProc Natl Acad Sci U S A
PubMed ID12422018
'The activity of the kinase Aurora-A (Aur-A) peaks during mitosis and depends on phosphorylation by one or more unknown kinases. Mitotic phosphorylation sites were mapped by mass spec sequencing of recombinant Aur-A protein that had been activated by incubation in extracts of metaphase-arrested Xenopus eggs. Three sites were identified: serine ... More
15-Lipoxygenase metabolism of 2-arachidonylglycerol. Generation of a peroxisome proliferator-activated receptor alpha agonist.
Authors Kozak Kevin R; Gupta Rajnish A; Moody John S; Ji Chuan; Boeglin William E; DuBois Raymond N; Brash Alan R; Marnett Lawrence J;
JournalJ Biol Chem
PubMed ID11956198
'The recent demonstrations that cyclooxygenase-2 and leukocyte-type 12-lipoxygenase (LOX) efficiently oxygenate 2-arachidonylglycerol (2-AG) prompted an investigation into related oxygenases capable of metabolizing this endogenous cannabinoid receptor ligand. We evaluated the ability of six LOXs to catalyze the hydroperoxidation of 2-AG. Soybean 15-LOX, rabbit reticulocyte 15-LOX, human 15-LOX-1, and human 15-LOX-2 ... More
Recombinant Dicer efficiently converts large dsRNAs into siRNAs suitable for gene silencing.
AuthorsMyers JW, Jones JT, Meyer T, Ferrell JE,
JournalNat Biotechnol
PubMed ID12592410
RNA interference (RNAi) is a powerful method for specifically silencing gene expression in diverse cell types. RNAi is mediated by approximately 21-nucleotide small interfering RNAs (siRNAs), which are produced from larger double-stranded RNAs (dsRNAs) in vivo through the action of Dicer, an RNase III-family enzyme. Transfecting cells with siRNAs rather ... More
Structure of a c-Kit Product Complex Reveals the Basis for Kinase Transactivation.
AuthorsMol CD, Lim KB, Sridhar V, Zou H, Chien EY, Sang BC, Nowakowski J, Kassel DB, Cronin CN, McRee DE,
JournalJ Biol Chem
PubMed ID12824176
The c-Kit proto-oncogene is a receptor protein-tyrosine kinase associated with several highly malignant human cancers. Upon binding its ligand, stem cell factor (SCF), c-Kit forms an active dimer that autophosphorylates itself and activates a signaling cascade that induces cell growth. Disease-causing human mutations that activate SCF-independent constitutive expression of c-Kit ... More
DEF6, a Novel Substrate for the Tec Kinase ITK, Contains a Glutamine-rich Aggregation-prone Region and Forms Cytoplasmic Granules that Co-localize with P-bodies.
AuthorsHey F, Czyzewicz N, Jones P, Sablitzky F,
JournalJ Biol Chem
PubMed ID22829599
Localization of DEF6 (SLAT/IBP), a Rho-family guanine nucleotide exchange factor, to the center of the immune synapse is dependent upon ITK, a Tec-family kinase that regulates the spatiotemporal organization of components of T cell signaling pathways and Cdc42-dependent actin polymerization. Here we demonstrate that ITK both interacts with DEF6 and ... More
The delta subunit of retinal rod cGMP phosphodiesterase regulates the membrane association of Ras and Rap GTPases.
Authors Nancy Vanessa; Callebaut Isabelle; El Marjou Ahmed; de Gunzburg Jean;
JournalJ Biol Chem
PubMed ID11786539
Post-translational modifications of GTPases from the Ras superfamily enable them to associate with membrane compartments where they exert their biological activities. However, no protein acting like Rho and Rab dissociation inhibitor (GDI) that regulate the membrane association of Rho and Rab GTPases has been described for Ras and closely related ... More
In Vivo Reconstitution of Saccharomyces cerevisiae DNA Polymerase epsilon in Insect Cells. PURIFICATION AND CHARACTERIZATION.
Authors Dua Rajiv; Levy Daniel L; Li Caroline M; Snow Peter M; Campbell Judith L;
JournalJ Biol Chem
PubMed ID11756442
DNA polymerase epsilon (pol epsilon) is a multiple subunit complex consisting of at least four proteins, including catalytic Pol2p, Dpb2p, Dpb3p, and Dpb4p. Pol epsilon has been shown to play essential roles in chromosomal DNA replication. Here, we report reconstitution of the yeast pol epsilon complex, which was expressed and ... More