scholarly journals Autoregulation of the Raf-1 serine/threonine kinase

1998 ◽  
Vol 95 (16) ◽  
pp. 9214-9219 ◽  
Author(s):  
Richard E. Cutler ◽  
Robert M. Stephens ◽  
Misty R. Saracino ◽  
Deborah K. Morrison
Keyword(s):  
Plasma Membrane ◽  
Protein Kinase C ◽  
Regulatory Region ◽  
Serine Threonine Kinase ◽  
Threonine Kinase ◽  
Rich Domain ◽  
Kinase C ◽  
Evolutionarily Conserved ◽  
Phosphorylation Event ◽  
Cysteine Rich Domain

The Raf-1 serine/threonine kinase is a key protein involved in the transmission of many growth and developmental signals. In this report, we show that autoinhibition mediated by the noncatalytic, N-terminal regulatory region of Raf-1 is an important mechanism regulating Raf-1 function. The inhibition of the regulatory region occurs, at least in part, through binding interactions involving the cysteine-rich domain. Events that disrupt this autoinhibition, such as mutation of the cysteine-rich domain or a mutation mimicking an activating phosphorylation event (Y340D), alleviate the repression of the regulatory region and increase Raf-1 activity. Based on the striking similarites between the autoregulation of the serine/threonine kinases protein kinase C, Byr2, and Raf-1, we propose that relief of autorepression and activation at the plasma membrane is an evolutionarily conserved mechanism of kinase regulation.

GFP-Tagged Protein Domains as Tools to Study Localized Second Messenger Function in Living Vells

1998 ◽  
Vol 4 (S2) ◽  
pp. 1014-1015
Author(s):  
T. Meyer ◽  
E. Oancea ◽  
T. Stauffer ◽  
M. N. Teruel
Keyword(s):  
Plasma Membrane ◽  
Protein Kinase C ◽  
Calcium Concentration ◽  
Protein Domains ◽  
Living Cells ◽  
Fluorescent Imaging ◽  
Protein Domain ◽  
Rich Domain ◽  
Kinase C ◽  
Cysteine Rich Domain

We have investigated the translocation of different GFP-tagged protein domains in response to various receptor stimuli. Confocal fluorescent imaging was used to study the localization of the GFP-tagged protein domain before, during and after the receptor stimuli was applied. A minimal cysteine-rich domain from protein kinase C was shown to be a useful tool to study localized diacylglycerol changes. Furthermore, the calcium-induced plasma membrane translocation of the GFP-tagged C2-domain from PKC could be used to study receptor-mediated changes in near plasma membrane calcium concentration. In addition, different pleckstrin homology domains (PH-domains) could be employed to measure changes in the concentration of phosphatidylinositol 4,5-bisphosphate (PI4.5P2) as well as of P13,4P2and PO,4,5P3. In combination, these fluorescent translocation probes provide a new means to investigate the local and temporal control of cell signaling processes in individual living cells.

scholarly journals CARMA1 Is Required for Akt-Mediated NF-κB Activation in T Cells

2006 ◽  
Vol 26 (6) ◽  
pp. 2327-2336 ◽  
Author(s):  
Preeti Narayan ◽  
Brittany Holt ◽  
Richard Tosti ◽  
Lawrence P. Kane
Keyword(s):  
T Cells ◽  
T Cell ◽  
Cell Receptor ◽  
Functional Interaction ◽  
Serine Threonine Kinase ◽  
Threonine Kinase ◽  
Kinase C ◽  
Evolutionarily Conserved ◽  
The Common ◽  
Protein Kinase C Activation

ABSTRACT Many details of the generic pathway for induction of NF-κB have been delineated, but it is still not clear how multiple, diverse receptor systems are able to converge on this evolutionarily conserved family of transcription factors. Recent studies have shown that the CARMA1, Bcl10, and MALT1 proteins are critical for coupling the common elements of the NF-κB pathway to the T-cell receptor (TCR) and CD28. We previously demonstrated a role for the serine/threonine kinase Akt in CD28-mediated NF-κB induction. Using a CARMA1-deficient T-cell line, we have now found that the CARMA complex is required for induction of NF-κB by Akt, in cooperation with protein kinase C activation. Furthermore, using a novel selective inhibitor of Akt, we confirm that Akt plays a modulatory role in NF-κB induction by the TCR and CD28. Finally, we provide evidence for a physical and functional interaction between Akt and CARMA and for Akt-dependent phosphorylation of Bcl10. Therefore, in T cells, Akt impinges upon NF-κB signaling through at least two separate mechanisms.

scholarly journals The cysteine-rich domain of human proteins, neuronal chimaerin, protein kinase C and diacylglycerol kinase binds zinc. Evidence for the involvement of a zinc-dependent structure in phorbol ester binding

1991 ◽  
Vol 280 (1) ◽  
pp. 233-241 ◽  
Author(s):  
S Ahmed ◽  
R Kozma ◽  
J Lee ◽  
C Monfries ◽  
N Harden ◽  
...  
Keyword(s):  
Protein Kinase C ◽  
Protein Kinase ◽  
Phorbol Ester ◽  
Hormone Receptors ◽  
Phorbol Esters ◽  
Steroid Hormone Receptors ◽  
Rich Domain ◽  
Kinase C ◽  
Cysteine Rich Domain

Diacylglycerol (DG) and its analogue phorbol 12-myristate 13-acetate (PMA) activate the ubiquitous phospholipid/Ca2(+)-dependent protein kinase, protein kinase C (PKC), and cause it to become tightly associated with membranes. DG is produced transiently as it is rapidly metabolized by DG kinase (DGK) to phosphatidic acid. Phorbol esters such as PMA are not metabolized and induced a prolonged membrane association of PKC. Until recently, PKC was the only known phorbol ester receptor. We have shown that a novel brain-specific cDNA, neuronal chimaerin (NC), expressed in Escherichia coli, binds phorbol ester with high affinity, stereospecificity and a phospholipid requirement [Ahmed, Kozma, Monfries, Hall, Lim, Smith & Lim (1990) Biochem. J. 272, 767-773]. The proteins NC, PKC and DGK possess a cysteine-rich domain with the motif HX11/12CX2CXnCX2CX4HX2CX6/7C (where n varies between 12 and 14). The partial motif, CX2CX13CX2C, is present in a number of transcription factors including the steroid hormone receptors and the yeast protein, GAL4, in which zinc plays a structural role of co-ordinating cysteine residues and is essential for DNA binding (protein-nucleic acid interactions). The cysteine-rich domain of NC and PKC is required for phospholipid-dependent phorbol is required for phospholipid-dependent phorbol ester binding, suggesting an involvement of this domain in protein-lipid interactions. We have expressed recombinant NC, PKC and DGK glutathione S-transferase and TrpE fusion proteins in E. coli to investigate the relationship between the cysteine-rich motif, HX11/12CX2CX10-14CX2CX4HX2CX6/7C, zinc and phorbol ester binding. The cysteine-rich domain of NC, PKC and DGK bound 65Zn2+ but only NC and PKC bound [3H]phorbol 12,13-dibutyrate. When NC and PKC were subjected to treatments known to remove metal ions from GAL4 and the human glucocorticoid receptor, phorbol ester binding was inhibited. These data provide evidence for the role of a zinc-dependent structure in phorbol ester binding.

scholarly journals KRAS interaction with RAF1 RAS-binding domain and cysteine-rich domain provides insights into RAS-mediated RAF activation

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Timothy H. Tran ◽  
Albert H. Chan ◽  
Lucy C. Young ◽  
Lakshman Bindu ◽  
Chris Neale ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Crystal Structures ◽  
Regulatory Region ◽  
Binding Domain ◽  
Published Data ◽  
Wild Type ◽  
Kras Mutations ◽  
Rich Domain ◽  
Structural Entity ◽  
Cysteine Rich Domain

AbstractThe first step of RAF activation involves binding to active RAS, resulting in the recruitment of RAF to the plasma membrane. To understand the molecular details of RAS-RAF interaction, we present crystal structures of wild-type and oncogenic mutants of KRAS complexed with the RAS-binding domain (RBD) and the membrane-interacting cysteine-rich domain (CRD) from the N-terminal regulatory region of RAF1. Our structures reveal that RBD and CRD interact with each other to form one structural entity in which both RBD and CRD interact extensively with KRAS. Mutations at the KRAS-CRD interface result in a significant reduction in RAF1 activation despite only a modest decrease in binding affinity. Combining our structures and published data, we provide a model of RAS-RAF complexation at the membrane, and molecular insights into RAS-RAF interaction during the process of RAS-mediated RAF activation.

Solution structure of a cysteine rich domain of rat protein kinase C

1994 ◽  
Vol 1 (6) ◽  
pp. 383-387 ◽  
Author(s):  
Ulrich Hommel ◽  
Mauro Zurini ◽  
Marcel Luyten
Keyword(s):  
Protein Kinase C ◽  
Protein Kinase ◽  
Solution Structure ◽  
Rich Domain ◽  
Kinase C ◽  
Cysteine Rich Domain

scholarly journals Identification of an alcohol binding site in the first cysteine-rich domain of protein kinase C δ

2006 ◽  
Vol 15 (9) ◽  
pp. 2107-2119 ◽  
Author(s):  
Joydip Das ◽  
Xiaojuan Zhou ◽  
Keith W. Miller
Keyword(s):  
Protein Kinase C ◽  
Protein Kinase ◽  
Binding Site ◽  
Rich Domain ◽  
Kinase C ◽  
Cysteine Rich Domain

scholarly journals KRAS interaction with RAF1 RAS-binding domain and cysteine-rich domain provides insights into RAS-mediated RAF activation

2020 ◽  
Author(s):  
Timothy H. Tran ◽  
Albert H. Chan ◽  
Lucy C. Young ◽  
Lakshman Bindu ◽  
Chris Neale ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Crystal Structures ◽  
Kras Mutation ◽  
Regulatory Region ◽  
Binding Domain ◽  
Published Data ◽  
Wild Type ◽  
Rich Domain ◽  
Structural Entity ◽  
Cysteine Rich Domain

ABSTRACTA vital first step of RAF activation involves binding to active RAS, resulting in the recruitment of RAF to the plasma membrane. To understand the molecular details of RAS-RAF interaction, we solved crystal structures of wild-type and oncogenic mutants of KRAS complexed with the RAS-binding domain (RBD) and the membrane-interacting cysteine-rich domain (CRD) from the N-terminal regulatory region of RAF1. Our structures revealed that RBD and CRD interact with each other to form one structural entity in which both RBD and CRD interact extensively with KRAS. Mutation at the KRAS-CRD interface resulted in a significant reduction in RAF1 activation despite only a modest decrease in binding affinity. Combining our structures and published data, we provide a model of RAS-RAF complexation at the membrane, and molecular insights into RAS-RAF interaction during the process of RAS-mediated RAF activation.

scholarly journals Evidence that a kinase distinct from protein kinase C and phosphatidylinositol 3-kinase mediates ligation-dependent serine/threonine phosphorylation of the T-lymphocyte co-stimulatory molecule CD28

1997 ◽  
Vol 326 (1) ◽  
pp. 249-257 ◽  
Author(s):  
Richard V. PARRY ◽  
Daniel OLIVE ◽  
John WESTWICK ◽  
David M. SANSOM ◽  
Stephen G. WARD
Keyword(s):  
Protein Kinase C ◽  
Protein Kinase ◽  
Cytoplasmic Tail ◽  
Jurkat Cells ◽  
Phosphatidylinositol 3 Kinase ◽  
Serine Kinase ◽  
Serine Threonine Kinase ◽  
Threonine Kinase ◽  
Kinase C ◽  
Phosphatidylinositol 3

The CD28 cytoplasmic tail contains several potential phosphorylation sites for the serine/threonine kinase protein kinase C (PKC) and/or proline-directed serine/threonine kinases, such as extracellular signal-regulated kinases. We demonstrate that ligation of CD28 by B7.1 results in strong serine/threonine phosphorylation of CD28. It is unlikely that ligation-stimulated phosphorylation of CD28 is mediated via activation of PKC, since it was not prevented by pre-treatment of Jurkat cells with inhibitors of PKC, and it was not mimicked by treatment with PKC activators such as PMA. Nevertheless, despite the lack of detectable effects of PMA treatment on CD28 phosphorylation, PMA did partially inhibit the association of CD28 with the putative signalling molecule phosphatidylinositol 3-kinase (PI 3-kinase) and the subsequent accumulation of PtdIns(3,4,5)P3. PI 3-kinase exhibits dual specificity as both a lipid kinase and a protein serine kinase, and site-specific mutagenesis of the Tyr173 residue in the CD28 cytoplasmic tail, which abolishes CD28 coupling to PI 3-kinase [Pages, Ragueneau, Rottapel, Truneh, Nunes, Imbert and Olive (1994) Nature (London) 369, 327–329], also prevents ligation-stimulated phosphorylation of CD28. However, the two PI 3-kinase inhibitors wortmannin and LY294002 had no effect on phosphorylation of CD28 after ligation by B7.1. This study therefore demonstrates that (1) a CD28-activated serine/threonine kinase distinct from both PKC and PI 3-kinase mediates ligation-stimulated CD28 phosphorylation, and (2) the PMA-stimulated down-regulation of the coupling of CD28 to PI 3-kinase is not due to PMA-stimulated phosphorylation of CD28.

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